Document MJanDoZJXBZrQOvmGODaXRnaV

fffi rTincPovrtf j5*eP',',u,# Topics: PCBs Chemical analysis Chamical reactions Chlorinated aromatic hydrocarbons Toxic materials Insulating oils EPRI CS-33C8 Protect 1263-11 Pinai Report Novamear 1983 State-of-the-Art Review: PCDDs and PCDFs in Utility PCB Fluid Prepared by SCS Engineers. Inc. Long Baacn, California MOMS 215169 State*of*the-Art Review: PCDDs and PCDFs in Utility PCB Fluid CS-3308 Research Project 1263-11 Finn Report Novemoer 1983 Prepared by SCS ENGINEERS INC 40u Long Beacn Boulevard Long Beach Calilorma 90807 Principal Investigators J Vuceta J R Marsn S Kennedy L Htidemann S Wiley Prepared lor Electric Rjwer Researcn institute 3412 Hiiiview Avenue Palo Alta California 9430* EPRi Protect Manager R V Komai Heat. Waste, and Water Management Program Coal Combustion Systems Division HONS 215170 ORDERING INFORMATION 'of C3(v#a of tnn r#oon snouia s* amectaa to o#se*rcn Peoorts Cant#r <RRCi 3ni siiion Pain A'fo 0*9*103 i*iS> 965-*08i Tnare * no cnarga tor'naoons raauast*n ov EPRl mnmner utilities and aftmatts u S utility associations U S government aaences if*dfl state *nn 'ocmi meoi* ana 'or*ion Orainriatton* **i|n wmcn R*I nas an ntormanon eimanoe sorem#m On reouest RPC win Sana a catalog of EPRl r-eoorts Cgovnem * tS*S E'oc*ne Nw a***arcn miniiil# i"c * 'ignit NOTICE *>* -won dwiih 6* ma siamtanowti namon oo<e** n an account oi woTM >oo' o* * i " e aowo* Anearcn inetitut# me 'EP*n uannor gppi nyrmi ot SPAT mt organieanorsii name ot-ow *or any ww tciine on oenarf 01 on* oi ntm iai mom an* warrant* tzoraaa or mo*ao wrin 'ofooci o -* oi an* miormaian aooa'aiut mo>noo or O'oeau oiacoaao m mu -oon or mat luC" *aa "** not "at 0' ia '* o**n#o r^ma Or 101 autumn an* laomnaa am rtaoect '0 mo uaa O' a' 'or oamaon 'eiuiiing "om u ai an* mtowaiion aoovaiut ntomoe or orocaaa ortcioaac * inn 'toon Aroaaran o* sea Enomaon me Long *Mcn Caworn.a HONS 215171 abstract An extensive literature search was conducted into the chemical, environmental, toxi cological, and analytical orooertles of chlorinated dioxins and dlbenzofurans, par ticularly as these relate to PCB fluids. The literature search olus consultations with the leading international researchers m the field was used to oreoare a timel state-of-the-art comoi lation of knowledge and theory on these important compounds. The chemistry of these compounds suggests that while chlorinated dibenzofurans can form directly from PCBs, chlorinated dioxins cannot. Both, however, can be formed from chlorophenols or chlorinated benzenes. Formation within electrical equipment PCBs under normal operating conditions is not likely. There has been little re search conducted into degradation techniques. Some chemical degradation techniques and photolysis show promise. These compounds are extremely persistent In the envi ronment, especially if protected from the sun. They are Immobile in soil and sedln*nt and do not bloma^ilfy to any great extent. On a molecular basis, 2,3,7,8-TCOO Is one of the most toxic synthetic molecules known to man, although Its toxicologi cal effects in man are stilt not well understood. Chemical analysis of dioxins and dlbenzofurans is still a developing science. All analytical nmthods rely on some form of gas chromatography/mass spectroscopy. A further development which shows same praeise as a rapid screening test is enzyme bioassay. HONS 15172 111 EPRI PERSPECTIVF PROJECT OESCRIPTIOK There has been inference and speculation about the potential presence and formation of polychlorinated dlbenzodloxms (PCOOs) and polychlorinated dlbenzofurans (PCDFs) in polychlorinated biphenyls (PCBs), This conjecture arises from the identification of these materials In combustion by-products of fires associated with PCS equipment. Th'ere has not been a consolidation or review of the pertinent chemistry and toxi cology literature from which one could separate legitimate scientific conclusions from unproven speculations and myths. This final report for RP1263-U endeavors to fill this need for a scientific litera ture review and serves as a literature basis on which subsequent experimental work may be developed. There are plans for a jointly funded project by two of EPRI's divisions--Electrical Systems Division and Energy Analysis and Environment Olvlslonto Investigate current analytic procedures, to analyze for specific PCDFs and PCODs in electrical equipment fluids, and to attempt to experimentally define the condi tions in electrical equipment that might lead to formation of PCDFs and PCOOs. PROJECT OBJECTIVES This project was conceived to examine and sunarize the state of the art of know ledge regarding PCOOs, PCOFs, and other cyclic hydrocarbons In utility PCB fluids. The study also was to determine what conclusions could be made about formation conditions and to define gaps In experimental data. PROJECT RESULTS There are a number of l^ortant conclusions from this study. These are: 1. The association of PCB equipment and fires leads to formation of PCDFs and PCOOs. 2. While PCBs can lead to PCOFs, the formation of PCOOs from PCBs is unlikely. 3. PCOOs can form from chlorophenols. HONS 216173 v 4. PCOOs can form from chlorobenzenes m air at temperatures oetween 130 and 620C. 5. PCDFs optimally form from PCBs at temperatures oetween 2/0 ano J00C and probaoly decomposes at temperatures above 3lXlwC. 6. In unused PCBs, PCDF levels as high as 20 ppm have been noted. 7. After a fire, levels of PCDCs are less than 5* of th# levels of PCDFs measured. 8. While sparking or soldering does not seem to cause PCDF formation froa PCBs, welding creates an increase of a factor of 1.4 to 5 times the original levels. 9. There are no data regarding fires and contaminateo mineral oil. This report is of Interest and value to utility and nonutlhty personnel wno are faced with the Issue of PCDFs and PCDDs in and around PCB equipment and wno neeo to know about analyses and toxicology. It is also useful to those who must respond to questions, since the report simmarlzes what is known and identifies areas wnere additional research is needed. A bibliography Is provided for those who wish further details. This information should provide sufficient background for those concerned about the presence of dioxin and dlbenzofuran In electrical equipment. The next step, wnicn will be addressed by other divisions at EPRI, will measure levels of PCOFs in old equipment and will attMpt to simulate extreme conditions under wnicn they might rorm. The Coal Combustion Systems Division plans to docimnt cleanup techniques that are used after a PCB fire. Ralph Y. Komal, Project Manager Coal Combustion Systems Division MONS 21517* ACKNOWLEDGMENTS This report on chlorinated dibenzofurans and dibenzo--d10xins could not nave oeen compiled without th* nd and cooperation of numtrous people. Chi of among tnetn ,,as Christoffer Rapp*, who provided data, figures, comnents, ana counsel, uthers wno have particularly aided in the task of Identifying pertinent tecnnical puolications and data sources, as well as providing expert comnents on tms subject, mcluoe Bengt Ahllng, Hans R. Buser, Gangadhar Choudhary, Paul Des Rosiers, Dennis Focnt, Anders Gara, Lennart Hardell, Otto Hutzinger, Lawrence H. Keith, Ernest Henan, and Thomas 0. Tleman. Providing the utility and EPRI perspective we thanx Gil Addis, Jacques Guertln, Harry Onlshl, W. Corey Trench, John P. Woodyard, and Walter Weyzen for their review comnents. Thanks also to the EPRI Project Manager, Ralph Kamel, for constructive guidance m formulating and executing tne project and presenting the Information. HONS 215175 vli CONTENTS Section 1 INTRODUCTION Sickground Project Objectives Report Organization 2 ACCIDENTS INVOLVING GENERATIONOF PCODs/PCOFs ANO OTHER RELATED CHLORINATEO HYOROCARBONS Introduction Electrical Power EquipmentEpisodes Nonelectrical Industrial Episodes Formation of PCDO, PCOF, and Other Chlorinated Organics 3 PCDO/PCOF/PCBP FORMATION Physical and Chemical Characteristics of PCD Os and PCOFs Cheanstry of Formation of PCODs and PCOFs Transfomtr/Cipacltor Chemistry 4 CHEMICAL ANO BIOLOGIC* DEGRADATION Chemical Deyedition Photolysis Incineration Biological Dey edition 5 ENVIROttCNTAL PERSISTENCE 6 PCOO/PCDF/PC0P TOXICOLOGY Introduction Animal Toxicity Human Toxicity Paoe 1-1 1-1 1-2 1-2 2-1 2-1 2*1 2-A 2*7 3-1 3-1 3-2 3-1* 4-1 4-1 *-3 4-5 4-7 5-1 6-1 6-1 6-1 6-15 MQNS 216176 ix CONTENTS (continued) Section 7 CHEMICAL ANO BIOASSAY ANALYSES Chemical Analyses Scope of the Analytical Problem Criteria for Selecting Analytical Procedures Sampling Methods Sample Extraction, Purification, and Separation Efficacy of Extraction and Cleanuo Methodology for PCDOs/PCDFs Instrumental Methods Quantification Confirmation Bioassays 8 RESEARCH NEEDS - Introduction Generation Toxicity Chemical and Bioassay Analyses Environmental Persistence Destructl on/Removal 9 REFERENCES APPENOIX A POLYCHLORINATED OIBENZO-P-OIOXIN CONGENERS BY MONOLOGUE ANO ISOMER (NlUBER OF ISOMERS) APPENOIX B POTENCY OF HALOGENATED BIPHENYLS IN INDUCING HEPATIC ARYL hyorocarbon hyqroxylase activity in chicken EMBRYOS APPENOIX C TRANSFORICR DATA FOR TESTS CONOUCTEDBY CHITTIM, ET AL. APPENOIX 0 EXTRACTION METHODS FOR PCODs ANO PCOFs APPENOIX E GLOSSARY ANO ABBREVIATIONS APPENOIX F MATRIX OF REFERENCES FOR PCBs Paoe 7-1 7-1 7-1 7-5 7-6 7.7 7-8 7-9 7-16 7-18 7-18 8-1 8-1 8-1 8-2 8-3 8-3 8-4 9-1 A-l B-l C-l 0-1 E-l F-l HONS 216177 X ILLUSTRATIONS 2-1 3-1 3-2 6-1 6-2 C-l C-2 C-3 C-4 C-5 Pertinent Chemical Structures Chemical Pathways to PCDOs, PCOFs, end PCBPs TCOF Isomer Distributions at Five PCS Fires The Most Toxic PCOO and PCOF Isomers PCOF Isomers Retained and Excreted in Yusho Patients Fragmentogram (Based on Mass Specific Detection) of a Fly Ash Sample (Above) and Standard Mixture of TCOO (Bottom) on a SS-m Sllar 10-C Column (l.d. 0.26 ne) (113) Kiltlple Selected Ion Mass Chromatogrmn Obtained for Calibration Mixture Containing the 22 TCOO Isomers (Column: 50-m Hybrid Phase MOOT Glass Capillary) (Brehm Laboratory, Personal Communications) Mass Fragmentograms (OV-17 Glass Capillary Column, 200-240'C) Showing Elution of PCODs m (a) Fly Ash of a Municipal Incinerator, (b) Fly Ash of an Industrial Heating Facility, and (c) Mlcropyrolysis Sample of Cortilned 2,4,6-trl-, 2,3,4,6-tetra, and pentachtoroonenate; m/e 320, 354, 3B8, 422, and 456 for tetra- to octa-CODs (34) Mess Fragmentogrmes (m/e 270, 304. and 338) of Pyroiyzed PentachloroPIpnenyls Stowing Elution of tri-, tetra-, and penta-COFs frae (a) 2,3,4,5,6-, (b) 2,4,5,2',5'-, (c) 2,4,S,3\41-, (d) 2,4,6,2',4'-, and (e) 2,3,6,2',5'pentechloroblphenyl (50-m OV-17 Glass Capillary Column, 100-160* at 207rin, 160-250* at 2*C/rin) (34) Mess Fragxentograms (m/e 304, 338, and 372) of Pyrolywd Hexachloroolphcnyls Showing Elution of tetra-, penta-, and hexa-COFs from (a) 2,4,5,2',4*.5'-, (b) 2,4,6,2*,4*,6*-, (c) 2,4,5,2\4',6'-, (d) 2,3,4,2',3',4`-, (e) 2,3,5,2',3'.5' (f) 2,3,6,2',3',6'-, and (g) 3,4,5,3`,4`,5'-hexachlorob1 phe nyl (50-m OV-17 Glass Capillary Column, 100-160* at 207min, 160-250* at 2*C/m1 n) (34) gaoe 2-2 3-3 3-13 6-2 6-19 C-2 C-3 C-4 C-5 C-6 HONS 215178 xl ILLUSTRATIONS (continued) Fj_2ure_ C-6 Mass Fragmentograms (m/e 338, 372, 406, and 440) of Pyrolyied Hepta- and Octachloroeiphenyls Snowing Elution of penta-, nexa-, hepta-, and octa-COF from (a) 2,3,4,5,2*,3', 4'-, (b) 2t3,4,S,2', 4',5'-nepta, and (c) 2,3,4,5,2' ,3',4' ,S`-octachloroblohenyl (50-m 0V-I7 Glass Capillary Column, 100-160* at 20*/rin, 160-250* at 2*C/mi n) (34) gaot C-7 HONS 215179 *11 TABLES Tab le 2*1 3*1 3-2 3-3 3-4 3-5 3-6 3- 7 4- 1 5- 1 5*2 6- 1 6-2 6- 3 7- 1 7-2 7-3 Summary of Chlorinated Aronatic Hydrocarbons Detected During Different Accidents [sowers of PCOFs and PCD Os Levels (ug/g) of PCDFs in Conwrciai PCBs PCDFs Found at Surahammar, Sweden PCDF Levels in PCB Fires PCBi and TCDF Concentrations (ppm) in Used Ask are Is Concentration of PCDFs In Askirel Transforner Fluids froa In-Service Distribution Transformers at GE Transformer Manufacturing Facl 11 ty pyrolysis of PCBs and Askarels at 500*C Combustion Criteria for PCBs and PCDF/PCDD-Contamlnated Wastes PCDD and PCDF Concentration Range In Hudson and Housatomc River Sediments and Fish from woods Pond Summary of Predicted Partition, Transfer, and Degradation Constants for 2,3,7,8-TCDD Biological Potency and Binding Affinity of Various Blphenylene and Biphenyl Congeners Suamary of the Ootage Associated with the Toxicological Criteria for Various PCDD Congeners Levels of PCOFs In Samples froa Yus ho Patients Instrumental Methods Applied to Detect PCDDs/PCOFs in Sample Extracts List of Extraction and Cleanjp Procedures Frequently Used for PCDD/PCDF Analyses Extraction Efficiencies of Polychlorinated Compounds from Fly Ash 2-8 3.2 3.9 3-12 3-12 3-16 3-17 3-13 4-6 5-3 5-3 6-11 6-15 6-18 7-2 7-7 7-ID MOMS 21Si80 xill TABLES (continued) Table 7-4 7-5 7-6 Influence of Time on Extinction Efficiency for PCODs end PCDFs Typicel Analytical Results for Quality Assurance Samples: Analysis of Fishfor2,3.7,8-TCDD Quantitative Analyses of PCOF and PCDD Isomers m Samples from PCS Incidents andIncinerator Fly Ash - Paoe 7-H 7-15 7-17 MONS 215181 xl v executive summary Since 1979, research has shown a ootential concern resulting from tht presence of chlorinated aronatlc hydrocarbons associated with PC8s, predominantly polychlori nated dibenzodloxins (PCOOs), polychlorinated dlbenzofurans (PCOFs), and other re lated compounds. These compounds occur in extremely low concentrations in PCS lifl uids. However, some PCOD and PCOF isomers are orders of maailtude more toxic than PCBs. There are a limited number of direct precursors and reactions by which PCOOs can be formed. In order to be a dioxin precursor, a compound mist neet two conditions: (a) the comoound must be an ortho-substituted benzene ring In which one of the sub stituents Includes an oxygen atom directly attached to the ring; and (b) it; mist be possible for the two substituents, excluding the oxygen, to react with each other to form an Independent comoound. The nmchanlsms by which PCOOs are formed during coabustlon are not well understood. Two suggested sources are (a) condensation of chlorophenols; and (b) thermal synthe sis from organic materials and Inorganic chloride (the "trace chemistries of fire* hypothesis). There has been some question regarding the role of PCBs in PCOO and PCDF formation. Since PCBs do not fit the precursor model, PCOOs are not expected to form readily in a mixture of biphenyl compounds. The analysis of soots resulting fron incidents in volving PCOOs and PCOFs Indicate the possibility of PCB conversion to PCDF or cnlorobenzene conversion to PCOO and PCOF In a malfunctioning transformer or capacitor, provided that there Is sufficient time at optimum temperature and/or oxygen condi tions for their formation. Ordinary working conditions do not appear to be condu cive to PCDO/PCOF formation. Electrical equipment PCB fires may begin internally or externally. If they begin Internally in an airless transformer or capacitor, the early stages are caused by electrical energy In Internal arcing. Under these conditions, the major combustion ES-1 MONS 215102 products are PCBPS; siillir amounts of PCOFs, PCPYs. and PCCYs are also found, [f tn equipmont explodos or cracks and tnt PCBs are exposed to air, higner concentra tions of PCOFs will form. If the fire Begins externally, PCOFs are tne major com bustion product, accompanied by PCPYs and PCCYs; no PCBPs Mill form. PCOOs will form only If cnlorobenienes/chlorophenols are present in the PCB or asxarel. The following cnemical degradation metnoos nave been successfully testeC on TCOD: oxidation by ozone, chlorine, or cnloroiooldes; alkaline dehydrochlonnetlon; cata lytic reduction with iron cnlorides; treatment by ruthenium tetroxide; and micellar catalysis using cnloroiodides. PCOOs and PCOFs are also susceptible to photolysis. Investigations have also shown that dioxins exniBit a relatively strong resistance to biodegradation. It appears that the predominant mode of elimination of TCOO In soils Is photodecomposition by sunlight, rather than microbial degradation. In aQueous ecosystems, 2,3,7,B-TCDD may be metabolized, but the rate is extremely slow. Little is known of the environmental behavior of most of these compounds; only 2,3, 7,8-TCDO has boon studied In any detail. In general, TCOO is highly persistent In soils, where It is protected from the Influence of solar photolysis and volatiliza tion. It is virtually Immobile in all but very sandy soils. The role of erosion as a transport mechanism is unknown. TCOO Is not taken up by plants to any significant degree, and there is relatively little Blomagnification in mammals. In water, TCOO is found to be strongly adsorbed onto bottom sediments. On a molecular basis, 2,3,7,B-TC00 is one of the most toxic synthetic molecules known to man. Both higher and lower chlorination reduce toxicity, but the toxic symptoms of the various isomers are similar, the only difference being the amount required to product a given effect. The PCOO and PCOF isomers with the highest acute toxicity are those with chlorines in at least the 2,3,7,8 positions. Most of the research to date has been conducted on the toxicology of 2,3,7,B-isomers in enimals. The toxic effects Induced by 2,3, 7.B-TC00 in all animal species included progressive weight loss and general weaken ing, with death occurring within a few weeks or months. 2,3,7,B-TCOO and other halogenated PCOOs exert their toxic effects by several mech anisms. They were primarily found to stimulate a number of enzyme activities (most MOMS 21Si3 ES-2 notably within the liver), primarily nepatic 6-ammolevulinic acid (ALA) synthetase and aryl hydrocarbon hydrolase (AHH). The PCOOs which have been found to induct ALA synthetase n*v two common properties: (1) halogen atoms occupy it least three of tne four ring positions (l.e., 2. 3, 7, 8); ind (2) it leist one free nonmlogemteo cirbon atom is unoccupied. 2,3,7,8-TCOO nis been found to be many qrders of magmtude more potent tnin my otner known inducer of ALA. As i result of tm virlous Occidents in which humans were exposed to PCOOs md PCOFs, som epidemiological information is ivulible on himn toxicity. Cnloncne. m often disfiguring ind persistent oeniitologicil disorder. Is the most common in. dicitor of poisoning In the maun subject. Chemical inilysis of PCOOs ind PCOFs Is i complex tisk, dlctited by (1) the Urge number of IsoMrs of eicn compound (l.e., 75 PCOO 1 sowers ind 135 PCDF 1 sowers), ino (2) the difficulty of sepenting ind identifying isewers within i given chlonnitlon cliss. In idoltlon, winy of these 1 sowers hive not been prepired In pure forw; even the prepired ones hive not been widely ivulible. Prerequisites for successful chewicil inilysis Include representitive sewpling ind good storige procedures; requireamnts for successful inilysis include efficient ex. tnction, siwple puri fleet ion (contilnwent), good sepention, ultnsensltive detec tion ind quintlficitlon, ind most desinble confirmation. There are several methods for extraction of PCOOs and PCOFs from different simple witrices. All of the methods entail the Initial addition of internal analytical standards, followed by destruction of the sample matrix (where practical) or extrac tion with an organic solvent, partitioning of the PCDDs/PCOFs into an organic phase and polar constituents into en aqueous phase and discarding of the litter, and liq uid chrMtographic cleanup (and, in some cases, HPLC cleanup) of the extracts. The cleanup steps remove matrix components which may Interfere with subsequent detection and quantification of PCOOs/PCOFs, such as lipids and most of the chlorinated hydro carbon residues. It is generally accepted that the only technique whicn has both the required sensi tivity and specificity for reliable detection and quantification of PCDOs/PCOFs Is coupled gas chramatography/mass spectrometry (GC/HS). The first attempt to apply mass spectrometry for quantitative determination of TCOO was the high-resolution mass spectromotric technique. The limitations of this technique are the inability HONS 215184 ES-3 td (a) (HitInsulin TCOO Ions from tnoso originating from certain PCBs, and (b) separate TCOO Isomers. Netnods using GC/1ow-resolution MS utilizes open-tubular packed gas cnromatographlc columns capable of separating TCOO from other PCOOs of different cnlorme content, but nave little capacity for separating discrete TCOO Isomers. Their specificity in isomer detenninations is derived from tne cleanup methods used. The combined GC (packed column)/nign-resolut1on mess spectrometrlc techniques are similar to tne nigh-resolution MS taennique, but nave better resolution and sensi tivity due to the nigh-resolution scanning over a narrower mass range and at a more rapid scan rate. A further enhancement of the capabilities of tne GC/Mgn-resolutlon MS technique was capillary GC columns. The use of capillary GC columns pro vided tne capability to separate discrete isomers of the PCOOs/PCDFs and improved the ability to obtain accurate quantitative data on the low concentrations of PCOOs/PCDFs In environmental samples. Considerable success has been recently achieved In the separation of tetra- and nexa-COO Isomers by HPIC fractionation of discrete Isomeric compounds. This tech nique appears to be particularly powerful when used In conjunction with GC/MS technlques, and does not necessitate the use of capillary GC columns. Confirmation of PCOOs/PCDFs can be based on the proper molecular Ions and fragments, as well as the Intensities of the Ion clusters due to the chlorine isotopes. If a sample contains a multitude of PCOO/PCDF isomers, the positive identification of PCOOs/PCDFs could be confirmed by cluster analyses. PCOOs, PCDFs, and PCBPs produce a similar and characteristic pattern of toxic lesions. Including hyperkeratosis and squamous metaplasia of the skin, and Induction of a number of coordlnetely expressed enzymes through stereospecific, reversible binding to a cytosolic receptor protein. These phenomena form the basis on which In vitro bloessey techniques have been developed for analyzing total PCOO/ PCDf biolo gical activity, which In turn reflects their toxic potency. The significance of the bioassays. In addition to assessment of toxicological effects. Is their usefulness as a rapid and reliable screen for detecting biologically active substances prior to chemical Identification and quantification. The two most extensively studied bioas say techniques are aryl hydrocarbon hydroxylase (AHH) Induction, and cell keratlnlzatlon. HOMS 215185 Future research is needed primarily into (1) the generation of PCDDs, PCOFs, end other related nalogenated aromatics in utility PCB fluids, (2) chemical and oioassay methods for their analysis, (3) their toxicities, (4) their environmental fate, and (5) procedures for tneir destruction. HONS 219186 ES-S Summary Since 1979, research has shown j potential concern resulting from the pretence of some chlorinated aromatic hydrocarbons associated with PC8s, predominantly poly chlorinated dlbenzodioxms (PCDOs), polychlorinated dlbenzofurans (PCDFs), and other related compounds, such as polychlorinated blphenylenet (PCBPs), polychlorinated pyrenes (PCPYs), and polychlorinated chrysenes (PCCYs). These compounds occur in extremely low concentrations In PCB liquids. However, some PCOO, PCOF. and PCBP isomers are orders of magnitude more toxic than PCBs. There have been a number of Incidents Involving these compounds. Including fires, industrial accidents, and contamination incidents resulting from Inappropriate waste disposal. Several of these incidents have Involved PCB-fllled electrical transform ers and capacitors. In each case, the electrical equipment wes caught in an Intense fire. The temperatures reached were extreme, far beyond the temperatures expected during normal (100* to 130*C), or even overload (200* to 250*C), conditions. Other Incidents Include a PCB-filled heat exchanger (in which the PCBs were again heated, probably In an oxygen atmosphere, for an extended period), an industrial accident involving the manufacturer of trichlorophenol, and an Instance of chemical contami nation from trichlorophenol manufacturing wastes. CHEMISTRY OF PCOOs AND PCDFs Very little published Information is available on the chemical and physical proper ties of chlorinated dioxins, dlbenzofurans, and related chlorinated cyclic hydrocar bons. They do not react with weak acids and bases and most redox agents, and are only slightly soluble in water and many organic solvants. In general, chemical sta bility Increases with increasing halogen content. There are a limited nueber of direct precursors and reactions by which PCDOs can be formed. In order to be a dioxin precursor, a compound oust meet two conditions: e The compound mst be an ortho-substituted benzene ring in which one of the substituents includes an oxygen atom directly attached to the ring. HONS 21*187 S-l  It must be possible for the two substituents, excluding the oxygen, to reect with eacn other to form an independent compound- Although these conditions limit the number of direct dioxin precursors, there are many indirect precursors, i.e.. compounds which do not fit the model, but can de grade or be transformed into direct precursors. The mechanisms by which PCODs are formed during combustion are not well understood; thus, the effects of combustion tmnoeraturr, fuel characteristics, catalysis, and other parneters have not been de termined. Two suggested sources are: e Condensation of chlorophenols. a Thermal synthesis from any organic materials and Inorganic chloride (the "trace chemistries of fire" hypothesis). Pyrolysis (620*0 of chlorobenzenes in the presence of air yields PCOOs. Since chlorobenzenes do not fit the conditions set down earlier for dioxin precursors, it was postulated that the alkaline hydrolysis of chlorobenzene to PCDO involves a twostep condensation process consisting of dleenzatlon of ortho-chlorinated phenoxy anions, followed by cycllzatlon of polychlorinated 2-phenoxy phenate. This dlemrization is exothermic. Below approximately 1B0*C, usually vary minor amounts of PCOOs {<1 ppm) are foreied, but significant Quantities (1 to 10 percent yields) result at temperatures between 180*C and 620*C. The high-temperature air oxidation of coal In the presence of chlorine, HCT, or haCl has been shown to yield PCOOs. However, coal is a comacrclal source of benzene and cumene (a phenol rew material), so that formation of chlorinated benzenes and phe nols Is a probable Intermediate steo. Thermal decomposition of lignin and polyvinyl chloride (typical constituents of municipal refuse) can yield phenol, cresol, cate chol, benzene, chlorobenzenes, HC1, etc. Thus, rather than just any organic mate rial serving as a potential precursor, It seems nsre likely that those compounds which can ba pyrolyied to benzene, chlorobenzenes, phenol, catechol, and/or inor ganic chlorine are more probable Indirect precursors. There has boon some Question regarding the role of PCBs in PCDD formation, [n 1981, an electrical fire In the State Office Building In Bln^iamton, hew York, resulted in the pyrolysis of PCB fluid from a transformer. Soot from this fire contained PCOFs and PCOOs. PCBs do not fit the precursor model discussed earlier, and PCOOs would not be expected to form readily In a mixture of biphenyl compounds. Conceivably, PCBs could be oxidized to polychlorinated hydroxyblphenyls, which could, in turn. S-2 HOMS 2&S1S8 cycllzt to for* dioxins. Another reaction that has been suggested is the degrade* tlon of PCBs to chlorinated benzenes, subsequent oxidation to chlorophenols, and cycllzatlon. However, neither mechanism quite fits the theraal decomposition pat tern for PCBs. The PCB fluid at Binghamton was a mixture of 65 percent Aroclor 125* and 35 percent chlorobenzenes. It was suggested that pyrolysis of the chlorobenzenes Is * more likely source of the PCODs than PCBs. This reaction was discussed earlier. While PCDFs and PCODs can share some of the same precursors, PCBs are the most com mon source of PCOFs. PCDFs have been detected at levels up to approximately 20 ppm in a number of unused Axmrlcan, European, and Japanese comucrclal PCB mixtures. Concentrations and Isomer ratios were found to vary with the type of PCB and the country of origin, probably as a result of differences in manufacturing conditions. Heating of PCB mixtures in air for 1 week resulted in the formation of PCDFs at tem peratures over 270*C. Maximum lewis were reached at 300*C, followed by declining levels at 330*C, suggesting decomposition. Primarily, dl- and trlchlorodlbenzofurans were formed in this experlHmnt. TCOFs found In used transformer askarels ranged from approximately 0.00* to *.7 ppm. The concentrations of TCOF In transformer askarels were found to Increase with time In service. Furthermore, the apparent changes In TCOF concentrations caused Oy dis charging or arcing were found to be negligible, most likely because of the short duration of the sparks or events causing failure. The amount of PCODs, If detected-, was usually less than 5 percent of the TCOF concentrations. Smell amounts of tarphenyl, guaterphenyl, and chlorinated terphenyl were also detected In some of the askarel samples, but they were not Quantified. Samoles of used oil taken from two capacitors (Prodelec 3010) were analyzed for PCOFs; no apparent Increase wes detected. The formation of PCOFs when capacitor oils are axpoaed to sufficient oxygen and high temperatures was Investigated by conduct ing laboratory tests with Prodelec 3010. The data Indicated that neither sparking nor soldering resulted In an Increase In the concentration of PCOFs; welding, howimr, resulted In a 1.4- to S-fold Increase In PCOFs. The above findings indicate the possibility of PCB conversion to PCOF or chloroben zene conversion to PCDD and PCOF In a malfunctioning transformer or capacitor, pro vided that there Is sufficient time at optimum temperature and/or oxygen conditions HONS 215189 $-3 for their formation, Ordinary wonting conaitions ao not appear to bo conducive to PCDD/PCOF formation. It snaulo also oe noted that most of me research on PCB/asxarel conversion has been performed using pure PC3s/askarel conversion has oeen per formed using pure PCBs or askarels containing at least 50 percent PCBs. rhe effects of dilution, as in contaminated mineral oil, nave not oeen fully explored. Fires in electrical equipment containing PCBs nave also resulted'ln PCOF formation. Soot and wipe samoles from PCB capacitor ana transformer fire* in tn* United States. Canada, Sweden, and Finland have all contained PCDFs. Electrical equipment PCB fires may oegin internally or externally. If they begin internally In an airless transformer or capacitor (with either no head space or an inert atmosphere), the early stages are caused by electrical energy in Internal arc ing. Under tnese conditions, tne major comoustlon products are PCBPs; smaller amounts of PCDFs, PCPYs, and PCCYs are also found. PCBPs were Identified for the first time In the aftermath of a 1981 capacitor explosion in Stockholm, Sweoen. If the equipment explodes or craexs a no the PCBs are exposed to air, higher concen trations of PCDFs will form. If the fire oegins externally, PCDFs are the major combustion product, accamoamed by PCPYs ana PCCYs; no PCBPs will form. PCODs will form only If chlorobenzenes/chlorapnenols are present in the PCB or askaret. CHEMICAL ANO BIOLOGICAL DEGRADATION Very little research has been conducted on oegradatlon mechanisms of dioxins ana di benzofurans. This Is particularly true of tne PCDFs; there is almost no information published on degradation. A number of researchers nave speculated on degradation pathways, but little actual rescarcn nas oeen conouctad. Several chemical degradation methoos nave oeen successfully tested on TC00. Ozone bubbled through a suspension of TCOD In water and caroon tetrachloride for 50 nours achieved 97 percent degradation. Chloroloaiaes have been found to be effective in laboratory experiments, but tests on actual soil samples have not oeen hignly suc cessful, Alkaline dehydrochlorlnatlon nas oeen tested on 2,4,5-trichloropnenol still bottoms containing up to 250 ppm TCOD. The reaction product was essentially free of 2,3,7,8-TCOO (ppt range) and otner chlorinated dioxins (i.e., more than 99,95 percent destruction). HONS 215190 S-4 Experiments were also conducted on tne separation of 2,4-0 esters from 2,4,5-T in Agent Orange By fractionation, using catalytic reduction with iron chlorides. Over heads were found to contain 93 percent 2,4-0 ester, wnereas Bottoms contained 93 percent 2.4.S-T ester. Rutnemum tetroxide was also found to destroy 2,7-DCQO and TC00 m laooratory tests. Micellar catalysis was tested using several chloroiodioes. The most promising results have been obtained with benzalkqnium dlchloroiodioe and cetylpyndinium dichloroiooide. An experimental stuoy was conducted to assess the feasibility of the chlorolysis process (l.e., chlorine gas as the oxidant at 600'C and 2500 pslg) to convert typi cal chlorinated solvent wastes and Agent Orange to useful products such as carbonyl chloride, hydrogen chloride, and caroon tetracnlor ide. Agent Orange initially con tained 14 to 18 ppm TCDD, whereas the carbon tetrachloride contained no measurable TCOD at a detection limit of I ppb. PCODs and PCOFs are also susceptible to pnotolysis. There are Basically three con ditions necessary for tne photoreductlon of chlorinated aromatic compounds: e Presence of hydrogen donor, usually in the solvent, e Ultraviolet (UV) li^it of appropriate wavelength, e Adsorption of tne llgnt. In sun llgnt. only those compounds absorbing UV llgnt aoove 290 nm are expected to be amnaole to pnotooegraoatlon. There nas been little research conoucted on tnermal destruction of PCDD. ana even less on PCOF. Olfferent researchers have reported 2.3,7,8-TCOO oestructlon at 80Q`C. In general, nowever. Incinerator operating conoitions currently considered adequate for TCOD destruction are a temperature of at least 1000*C and a dwell time of 2 seconds. At 700*C and a dwell time of 21 seconds, only SO percent destruction was achieved; at 800'C and a dwell time of 21 seconds, 99.5 percent destruction was achieved. Investigations nave snown that dioxins exmoit a relatively strong resistance to biodegradation. Mott of the wont on oioxins nas been rocuseo on 2,3,7,8-TCOO be cause of Its extreew toxicity. Approximately 100 strains of microoes, wnicn nao pre viously shown the ability to degraoe persistent pesticides, nave oeen tested. Only five strains were shown to have the ability to degrade 2,3,7,8-TCOD. It is possible MONS 215191 S-5 that the lesser chlorinated PCODs ire more susceptible to biodegrsdition, as is the case with PCBs. It also appears that the predominant mode of elimination of TCDD in soils Is photodecomposltlon by sunlight, rather than microbial degradation. In aQueous ecosystems, 2,3,7.8-TCOD may be metabolized, but the rate is extremely slow. ENVIRONMENTAL PERSISTENCE With their high toxlcltles. the envirormental fates of PCODs and PCDFs are of In creasing importance. However, little is known of the environemntal behavior of most of these conpounds. Only 2,3,7,8-TCOO has been studied In any detail, often with fnconclusive results. Primarily as a result of the 1976 PCDD Incident m Seveso, Italy, most of the envi ronmental research has centered on the fate of TCOO in soil. In general, TCOD is highly persistent in soils, where it Is protected fron the influence of solar photo lysis and volatilization. It Is virtually immobile in all but very sandy soils. The role of erosion as a transport mechanism is unknown. TCOO Is not taken up by plants to any significant de^-ee, and there Is relatively little biomagmfIcatlon in mammals. In water, TCOO is found to be strongly adsorbed onto bottom sediments. TOXICOLOGY On a molecular basis, 2,3,7,8-TCOO Is perhaps the most toxic synthetic molecule known to men. Only bacterial exotoxins (l.e., Botulinum Toxin A, tetanus toxin, and diphtheria toxin) are more potent poisons. Both higher and lower chlorination reduce toxicity, but the toxic symptoms of the various isomers are similar, the only difference being the amount required to pro duce a given effect. The Isomers with the highest acute toxicity are 2,3,7,8- tetra-COO, 1,2,3,7.8-pentaC00, 1,2,3,4,7,8-, 1,2,3,6,7,8-, and 1,2,3,7,B,9-hexa-C0D. 2,3,7,B-tetra-CCF, 1,2,3, 7.8- and 2,3,4,7,8-ptnta-CDF, and 1,2,3,4,7,B- and 2,3,4,6,7,8-hexa-CDF. Host of the research to date has been conducted on the toxicology of 2,3,7,8-isomers in ani mals. The toxic effects Induced by 2,3,7,8-TCDD in all animal species included progresslve weight loss and general weakening, with death occurring within a few weeks or months. However, various experiments conducted to date have failed to provide the exact biochemical mechanisms of 2,3.7,8-TCOO lethality. MGNS 21S192 2.3.7.8- TC00 and otner halogenated PCODs were found to stimulate a number of enzyme activities, most notably within the liver, Heoati c a -anrmolevulime acid (ALA) syn. thetase and aryl hydrocarbon hydrolase {AHH) Mere found to be induced by the ores* ence of PCDDs, particularly 2,3,7,8-TCDD. The PCDDs which have been found to induce ALA synthetase appear to have two cannon oroperties: (1) halogen atoms occupy at least three of the four ring positions (i e., 2, 3, 7, 8); and (2) at least one free nonhalogenated carbon atom is unoccupied. Available toxicological data Indicate that those PCDDs that are not toxic generally do not Induce ALA synthetase; those which are lethal, even at extremely low concentrations, induce the enzyme. 2,3,7,8TCOD has been found to be many orders of "latitude more potent than any other known inducer of ALA. Microsomal monooxygenase (e.g., AHH) is a membrane-bound enzyme complex which cata lyzes the oxidative (and for certain substrates, reductive) metabolism of numerous xenoblotics and endogenous lipophilic compounds. 2,3,7,8-TCDD is a highly lipo philic macromolecule. The Induction of AHH and ALA by various PCODs are carried out by the same mechanism. Theoretically, the most logical pathway of 2,3,7,8-TCDD netabolism is by hydroxylatlon at the 1, 4, 6, or 9 position of the molecule by AHH. The hydroxylation of aromatic rings in positions adjacent to halogen atoms Is an uncommon reaction of the cytochrome P-450 monooxygenase system. This most likely accounts for the netabolic stability of 2,3,7,8-TCDD within mammalian systems. However, it has been postulated that 2,3,7,8-TCDD moves Into a cell and binds to a specific cytosolic receptor. The receptor-dioxin complex then moves into the nucleus of a cell, where it initiates the synthesis of a specific messenger ANA, which directs the synthesis of cytocnrome P-450. Other 2,3,7,8-TCDO molecules can then react with newly formed cytochromes, possibly to produce reactive Inteneediates that reduce or impair cellular metabol ism, gradually inducing cell death. These metabolites may be excreted as innocuous by-products which msy affect specific cells in other organs, or may act as carcino gens or cocarclnogens. If the activity of AHH were Increased, more 2,3,7,8-TCOO would be metabolized. If 2.3.7.8- TCDD, rather than a netabollte, is responsible for the toxicity, then in creased metabolism by the P-450 monooxygenase system should lead to a decrease m toxicity. Conversely, if a netabollte(s) is responsible for the toxicity of 2,3,7,8TCDD, then an Increase In metabolism may result In an Increase in toxlcl-ty. Several "ONS 21*193 S-7 studies nm show that 2,3,7,8-TCDO induces many enzyme systems while suppressing others. In addition to ALA and AHH, 2,3,7,8-TCOO induces tne following enzyme*: e UQP olucuronyl transferase, e Aloenyoe denyorogenase. e Glutatmone transferase S. e 0T*d1 apnarase. e Benzopyrene nyorolase. e Glutathione S-transferase. e Etnoxycaumarin deetnylase. 2,3,7,8-TCOO reduces the activity of the following enzymes: e UOP-glucuronic acid pyrgonospnatase. e D-glucuronolactone denyorogenase, e L-gluconate cenyorogenase. Anotner mecnanism by which 2,3,7,8-TCDO may exert its toxic effect is by the in crease or llpofuscin pigamnts. Lipid peroxidation, whlcn precedes the formation of polymeric llpofusclns. Is know to seriously damage memoranaus subcellular organ elles, including lysosomes. In one or more species of laboratory animals, bone marrow nypoplasia, testicular de generation, renal pelvis ano urinary bladaer nyperplasla, and nemorrnage in the in testines and aarenals nave oeen observed. Chronic administration of low levels of 2,3,7,8-TCOO to rats increases the incidence of neoplasia (tumors). The most malig nant tumors were observed In tne lungs, kioneys, liver, and sxeietal musculature. Otner neoplasms were found In the brain, skin, testes, ano ears. 2,3,7,8-TCOO Is a teratogen, interfering witn only a few emoryonic or fetal davelaonental processes. These teratogenic effects Include impacts on hormones involved in reproduction, thymic atroohy, fatty infiltration of the liver, general edema. delayeo head ossification, low birth weitt, fetal reaosorptlon, and kidney abnormali ties. As a resuit of the various accidents in which humans were exposed to PCDOs and PCDFs, some epidemiological information Is available on human toxicity. Chloracne, HONS 211194 S-8 an often disfiguring ano persistent dermatological disoroer, is tne most cannon in. dicator of poisoning in tne human suoject. However, the aosence of chloroene ooes not oreclude intoxication. Liver dysfunction is manifested by nepatomeoaly ano elevation of enzyme levels. Daily human exoosure to 0.D1 ug of 2,3,7,8-TCQO may result in Incipient carcinopen 1 city, but existing evidence indicates tnat PCDDs are at most only mildly carcino genic. Exposure to 4 ug of 2,3,7,8-TCDO would result in a shortened lifespan, and daily exposure to 290 ug would likely cause acute toxicity. The lmmeoiate symptoms of exposure to dioxins mcluoe a ourning sensation in one eyes, nose, and tnroat, fclloweo by neaaacnes, dizziness, nausea, ano vomiting. Several days later, severe itching, reoness, and swelling of the face (more marked over the eyelids, nose, and lips) may develop just prior to tne onset of chloracne. It has been snown tnat as little as 20 ug of 2,3,7,8-TCOQ on tne skin can lead to chloracne. Within tne initial weexs after exposure, inflamed nooules as well as pustules appear on tne face, forearms, shouloers, necx, and trunx, leaomg to come dones ano cysts. Sometimes chloracne is preceded by erytnenatous ana edematous sxi lesions. After 1 or 2 montns. acneform eruptions e*rge, and the skin Decomes hyperpignmnted. At approximately the same time, muscles, primarily in tne tnigns ano cnest area, become aggravated upon exertion. Insonnia, extreme fatigue, ner. vousness, irritabi 11 ty, and loss of libido also occur during this period. Otner symptoms of exposure include artnralgias. porpnyria cutanea tarda, nyperlipi. demia, cutaneous nyperpigmentatlon, and hirsutism. Porpnyria cutanea tarda, a sxm condition tnat usually occurs as a photosensitive dermatosis, is causeo by tne oe* velopment of vesiculooullous lesions over exposea areas. The oermatosis is precioi tateo oy minor trauma, ano may result in areas o* nealeo oullae, crusts, scars, and mi ii a. Blurred vision ano tne loss of taste and smell may also occur. There is no suostantiue evidence to indicate mutagenic, teratogenic, or emaryotoxic effects on fetuses from 2,3,7,8-TCDO exposure in vivo, however, cnranosomal abnor malities nave oeen reported in in vitro cytogenic stuoies on numan lynenocytes ex pos eo to 2,4,5*T containing 0.09 ppet 2,3,7,8-TCDO. Chromatid breaks ano oeletions mcreaseo witn increasing concentrations of 2,4,5-7. It was not possible to oetermme wnetner this was a toxic or genetic effect. HONS 245195 S-9 Analyse! of liver, adipose tissue, and blood samples of both Japanese and Taiwanese patients suffering from rice oil disease ("Yusho") indicated that the hipest con. centrations of PCDFs were found in the liver. PCOF isomers, which were metabolized and excreted fairly rapidly (e.g., 2,3.6,7-tetra-, 2,3,4,6,7-penta-, 1,2,6,7,3penta-, 1,2,3,4,8-penta-, and 1,2,3,4,6,7-hexa-CDFs), nave two vicinal unchIormatea C-atoms in at least one of the two aromatic rings in the PCOF nucleus. In contrast. PCOF isomer! retained in the bodies, such as 2.3,6,8-tetra*, 2.3,7,8-tetra-. 1,2,4, 7,8-penta, 2,3,4,7,8-oenta, 1,2.3,7,3-penta, 1,2,3,6,7,8-hexa-, and 1,2,3,4,7,3hexa-CDFs, have lateral positions substituted with chlorine. The complete elimi nation of this compound from the organism is a function of its capacity to change macrmolecules to more polar molecules. In general, the clinical appearance of contaminated animals and the variety of lesions found during post-mortem examinations are identical for TCDFs, 2,3,7,8TCODs, and PC8s. The toxicity of TCOF is approximately 10 percent that of 2,3,7,8TCOO, and 1000 times that of Aroclor 1242. The correlation of nmtabollsm with de toxification Indicates that the greater toxicity of 2,3,7,8-TC00 versus TCOF could be the result of the relatively slower nmtabollsm and excretion of 2,3,7,S-TC00. Studies indicate that TCOF nmtabollsm Is earned out through a detoxification mech anism; speclts that are able to nmtabollze and purge TCOF are more resistant to its acute toxicity. The multiple-organ and histologic damage may be a result of both the relative rate of formation and the further inactivation of the reactive metabol ites in different organ! (e.g., liver, kidneys, stomach, etc.). CHEMICAL ANALYSES Chemical analysis of PCODs and PCDFs Is a complex task, dictated by (I) the large number of isomers of each compound (i.e., 75 PCOO isoamrs and 135 PCOF isomers), and (2) the difficulty of separating and Identifying Isomers within a given chlorination class. All classes are quite similar structurally, and thus similar in chemical and physical properties. In addition, many of these isomers have not been prepared m pure form; even the prepared ones have not been widely available. To date, the fol lowing Isomers have been synthesized: the entire sets of tetra-, hexa-, hepta-, and octa-COOs; 2 dl-, 4 trl-, and 14 penta-CODs; all tetra-COFs; and some penta-COFs. Many of the early analytical studies focused on the determination of 2,3,7,3-TCOD because of Its extreme toxicity. In addition, this PCOO Isoner was one of the ear liest available in sufficient ouantlty to use as an analytical standard. Some of this nmthodology was gradually adapted, modified, and expanded to accommodate HONS 215196 s-to analyses of other PCODs and PCDFs. although entirely new methods have also been developed. Prerequisites for successful chemical analysis include representative sampling 4no good storage orocedures; requirements for successful analysis include efficient ex traction, sample purification (containment), good separation, ultrasensitive detec tion and quantifIcation, and most desiraote conf 1 rmation. Sampling Methods Sampling nathods can markedly affect the results obtained. In general, the follow ing dictates of good analytical practice should be applied in sampling: homogeneity of the sample matrix; collection of a truly representative sample; statistical con siderations with respect to replicate analyses; and the resultant effects on preci sion and accuracy. In cases where samples are acautred by simpte grab samp lino, such considerations can usually be applied. In other instances (e.g., collection of flue gas samples from incineration sources), the sampling procedures are far less standardized, and their effectiveness and limitations have not been adequately eval uated. Sample Extraction, Purification, and Separation There are several methods for extraction of PCODs and PCDFs from different sample matrices: U.S. Environmental Protection Agency (CPA): fish, adipose tissue, milk, water, soil, sedlnents. U.S. Food and Omg Administration (FDA): chemical formulations, fish. U.S. Fish and Wildlife Service: fish. Dow Chemical Company: fish, milk, fly ash, dust, urban particu lates, municipal sludge, soil. e Wright State University - Brehm Laboratory: soil, chemical waste, aqueous effluents, particulates. Umea University, Sweden: PCBs, fly ash, soot, wipe tests, blood. All of the above methods entail the initial addition of internal analytical stan dards, follOMd by destruction of the sample matrix (idiere practical) or extraction with an organic solvent, partitioning of the PCDDs/PCDFs into an organic phase and polar constituents into an aaueous phase and discarding of the latter, and liquid S-ll MONS 21*197 chromatographic clearejp (and, in some cases, HPLC cleanup) of the extracts. The cleanuo steos are intended to remove matrix comoonents, such as tipids, as 11 as most of the chlorinated hydrocarbon residues in the ssnole (e.g., PC8s, DOE. otner common chlorocarbon residues, and fused-ring aromatic compounds), which may inter, fere with subseouent detection and ouantification of PCODs/PCOFs. Instrumental Methods Instrumental nmthods which have been utilized for detection and/or ouantification of PCDDs/PCDFs m different samole matrices are as follows: e UV Soectroscooy - TCOO. e Thin Layer Chromatograohy - biological tissue; oils. e Electron Caoture/Gas Chromatography (EC/GC) - chemical formulations; PCOOs. e Probe Mass Spectrometry (MS) - milk; animat tissue. e Thermally Modulated EC Oetector {TMECO) - animal tissue; chemical formulations. e Packed Column GC/Low-flesolutlon MS (Selected Ion Monitoring) chemical formulations; industrial hygiene saaoles. e Packed Column GC/Hi oh-Resolution MS (Selected Ion Monitoring) . animal tissue; chemical wastes; aaueous solutions; soils; plant material. e Capillary Coluim GC/High-Resolution MS (Selected Ion Monitoring) human milk, animal tissue; water; sedlitents; combustion products. e Capillary Coluim GC/Low-Resolution MS (Selected Ion Monitortnq) . combustion products; oils; PCBs; human tissue. e Liquid Chromatography Packed Coluim GC/Low-Resolution MS (Selected Ion Monitoring) . combustion products; Isoamr separation and identification. e GC/Negative Chemical Ionization MS - animal tissue; wood; human tissue; combustion products. e GC/Negative Ion MS animal tissue; sediments. e GC/Atmospheric Pressure Ionization MS animal tissue; identifies, tlon of isomers in standard mixtures. Methods such as UV spectroscxy, thin layer chromatography, GC/EC detection, and TMECO have been developed for rapid screening of samples or qualitative detection. HOMS 215198 S-12 It u general 1y accepted that the only technioue which has both the required sensi tivity and specificity for reliable detection and ouantificatlon of PCDDs/PCOFs is coupled gas chromatography/mass soectronetry (GC/MS). The first attempt to aoply ness soectrometry for Quantitative determination of TCOO was the high-resolution mass soectronetric techmoue. High-reso lution scanning was accomplished over a nar row mass rang* (typically 0.3 AMU) at a rapid rate (4 scans/secoffd). dpt to the relatively high mass soectral resolution achieved, this technioue wet edeeuate for the purposes of distinguishing the molecular ion of TCOO from Ions arising from the sample matrix (mostly biological samples), however. It was soon realized that tne TCOO Ions could not be distinguished from those originating from certain PC8s, if the latter were present in the sample extract tn large concentrations relative to the TCOO (ppm versus ppt). Another limitation of the latter method was its inabil ity to seoarate TCOO isomers. GC/low-resolution MS utilizes open-tubular packed gas chromatograohlc coluims. This method was caoable of seoaratlng TCOO from other PCODs of different chlorine con tent, but had little capacity for separating discrete TCOO Isomers. The detection limits achieved in these determinations cre tn the range of 0.001 to 0.1 pom. Their specificity In Isomer determinations was derived from the cleanjo methods used. Furthermore, these mthods reoulred that the analyte exhibit the aooropriate GC retention tin and a mass spectral resoonse at the aooraorlate Ion masses corre sponding to TCOO. The combined GC (packed colour)/h1gh-resolution mass spectrometric techniques were developed for determining TCOO In beef tissues and related environmental samoles. These mass spectrometric nethods were similar to the high-resolution MS technioue, but were superior In that hlgh-resolution scanning was generally accomotished over a narrower mass range and at a more rapid scan rate. This improved both the resolu tion capability and the sensitivity of the technique. A further enhancement of the capabilities of the GC/hlqh-resolution MS techmoue for analysis of PCDOs/PCOFs entailed the Incorporation of capillary GC coluons. The use of capillary GC coluims provided the capability to seoarate discrete isomers of the PCOOs/PCOFs. The use of capillary coluim GC/hlgh-resolution MS also improved the ability to obtain accurate quantitative data on the low concentrations of PCODs/ PCOFs In environmental samples. HOMS 215199 S-13 Several laboratories have increased the use of capillary coluim GC/1ow-resolution MS in order to provide data on a wide range of PCD D/PC OF isomers m various types of samples, including PCS electrical fluids. Considerable success has been recently achieved in the seoaration of tetra- and ;iexa-COO isomers by HPLC fractionation of discrete isomeric cmoounds. This tech* nioue aopears to be particularly powerful when used in conjunction with SC/MS tech, mgues, and does not necessitate the use of capillary SC colours. No beta on seoara. tion of PCDF i soamrs by this method have been reported. decently, several newer mass spectrometnc techniques have also been investigated for the analysis of PCODs/PCOFs. Among these are negative ion MS, negative ion chemical ionization MS, and atmospheric pressure/ionization MS. Results obtained thus far show that all of these offer enhanced sensitivity, selectivity, and soecif icity. Quantification To date, quantification has been based on peak area measurements by conparison with either internal standards or a calibration curve of known quantities of authentic standards {external standards). It has generally been assumed that all isomers of a particular PCOD or PCOF have the same response using the MS quantification. Two re cent studies on the responses of 28 and 20 different PCOF isomers both showed a 3 fold variation in the response using an electron impact (El) mode. Aoplication of the negative chemical ionization (NCI) mode, however, resulted in 20-fold and 25 fold variations, respectively. For the higher chlorinated homologues, the differ ences appeared to be much smaller. Confi^nnation Confirmation of PCODs/PCOFs can be based on the proper molecular ions and fragments, as well as the intensities of the ion clusters due to the chlorine isotopes. More over, if a sample contains a imiltitude of PCDD/PCDF isomers, the positive identifi cation of PCODs/PCOFs could be confirmed by cluster analyses. For adequate confirmation of PCODs/PCOFs, there are several parameters available. Compared to an authentic standard, the PCODs/PCOFs in the sample should have the same following parameters: retention volume in the coluim chromatooraphy in the cleanup steps; retention time in the gas chromatographic separation; molecular HONS 15200 S-14 Ktight; fragmentation and chlonne cluster in the MS detection; and difference in response factors comparing the El and the NCI modes. These parameters, in addition to access to wel 1-defmed weighted standards, give the possibility of rei^le analyllS of PCDDs/PCOFs. bioassay analyses -" as discussed earlier, numerous clinical biomedical parameters have been shlwi to be altered m animals and humans recewi ng different dotes of PCDOs, PCOFs, *tf PCBPs. In a number of investigations, 2,3,7,8-TCOO served as a prototype of Individual PCOD and PCDF congeners and their mixtures. All of these conpounds produce a similar and characteristic pattern of toxic lesions, including hyperkeratosis and squamous ireta* plasia of the skin, and induction of a nuxber of coordinate^ expressed enzymes through stereospecific, reversible binding to a cytosolic receptor protein. These phenomena form the basis on which In vitro bioassay techniques have been develooed for analyzing total PCDO/ PCDF biological activity, which in turn reflects their toxic potency. The significance of the bioassays. In addition to assessment of toxicological effects, Is their usefulness as a rapid and reliable screen for detecting biologi cally active substances prior to chemical Identification and quantification. The discussion that follows focuses on two of these bioassay techniques: aryl hydrocar bon hydroxylase (AHH) induction, and cell keratlnlzatlon. Aryl Hydrocarbon Hydroxylase fAHH) Induction The monooxygenase enzyme system, consisting of NAOPH-cytochrome P-450 reductase and cytochrome P-450, 1$ an Integral part of the smooth endoplasmic reticulum, and cata lyzes the catabolism of lipophilic compounds to more polar mtabolltes. The Induction of the hepatic monooxygenases by 2,3,7,B-TC00 occurs In most tissues of most species. These bioassays are extrewly sensitive to a variety of chlori nated aromatic hydrocarbons. A large number of specific enzymes have been assayed; the most extensively studied of them Is AHH. AHH activity Is a measure of cytochrome P-448. The induction of AHH 1$ mediated by the reversible stereospecific binding of haloginated aromatic hydrocarbons to a cytosol protein, the cytosol Induction receptor, idtlch mediates the nuclear uptake of the toxicant. The receptor is determined by the Ah locus In mice. The Ah locus, MOHS *15301 S-15 and hence the cytosol receptor, controls not only the induction of AHH activity but also a battery of coordinatety expressed enzymes. Two lines of evidence suggest that toxic lesions oroduced by TCOO, as well as bio chemical responses, are mediated by the cytosolic receptor. First, the binding affinities of a large number of halogenated aromatic hydrocarbons for the receotor correspond very closely with the ranking of tneir toxic potencies, SfteOftd, two -'xlc responses produced by TCOO in mice, thymic Involution and cleft palate forma tion in fetuses, have been shown to segregate with the Ah locus. Compared with classical polycyclic aromatic hydrocarbon Inducers of AHH, such as 3methylcholanthrene, 2,3,7,8-TCOO produces a parallel dose-response curve; both com pounds produce the same maximum response In AHH activity. However, 2,3,7,8-TCOO is g 3 x 10 times more potent than 3-methylcholanthrene as an AHH Inducer. The AHH Induction bioassay utilizes a rat hepatoma cell line which possesses low basal AHH activity and is highly inducible for this enzyme. The amount of AHH in. ductlon Is determined by the fluorimetrlc assay of 3-hydroxybenzo[a]pjrene formation from benzo[a]pyrene (the substrate for the AHH enzjmm assay), or by the fluorimetrlc determination of 7-ethoxyresorufln 0-deethyl at ion. The binding of a specific ligand or mixture of ligands Is determined by nmasurlng the dose-dependent displacement of [^H]-2,3,7,8-TCOO previously bound to the protein. Toxicity to Cells Cultured In Vitro One of the most characteristic responses to TCOO Is hyperkeratosis of the skin. When XB cells derived from a mouse teratoma are cultured at high density (to prevent spontaneous dlffentlatlon) along with lethally irradiated 3T3 cells, the addition of TCOO produces a dose-related keratlnlzatlon response, as detected by red staining with Rhodanlle blue. This Is a direct effect of TCOO on X8 cells, and will occur in the absence of the feeder cells if the teratoma cells are cultured in 3T3-condltloned kmdla. XI cells contain the cytosol receptor, and respond to TCOO with a dose-related Induction of AHH activity. Other halogenated aromatic hydrocarbon congeners, including PCODs, PCOFs, PCBPs, and uobenzenes, were also found to induce keratlnlzatlon although to a lesser extent than 2,3,7,8-TCDD. Keratlnlzatlon could also be induced by some normalooenated aro matic hydrocarbons, such as benzanthracene and 5,6-benzoflavone. Since 2,3,7,8-TCOO Is the most potent, but not the only Inducer of this hyperkeratlnlzatlon response. HONS *1*202 S-16 this effect is referred to as "dioxin-like" activity. Unrelated toxins, however, failed to produce keratinization in the X8/3T3 culture system. Furthermore, a cor relation has been demonstrated between the degree of toxicity of various TCOO isoiers and congeners and the extent of hyperkeratin 1 lation response. The X8/3T3 system is extremely sensitive, inducing keratinization with as little as 3.2 pi cograms of TCOO. In addition, this system is particularly relevant as a modal, since it deals with an induced differentiation of the Intact cell using an endpoint (hyperkeratinization) known to be relevant to human exposure (i.e., chloracne). RESEARCH needs Future research is needed primarily into (I) the generation of PCDDs, COFs, and other related halogenated aromatics in utility PCB fluids, (2) chemical and bioassay methods for their analysis, (3) their toxicities, (4) their environmental fate, and (5) procedures for their destruction. Generation of PCOPs, PCOFs, and Related Compounds Since available information on both conditions and quantities of formation is insuf ficient anq often contradictory, future studies should focus on better delineating the generation of PCDFs, PCDDs, and PCBPs. Standard and non-standard operating con. ditions should be compared with optimum formation conditions to determine if forma tion is indeed possible, and if the necessary precursors exist m PCS fluids. Such tests would provide specific insights into the chemistry, mechanisms, and ootmum reaction conditions favoring the formation of these compounds from PCB fluids. The data on structure-biological activity relationships for 2,3,7,8-TCOO m labora tory animals are extensive, while other PCOO homologues have been studied to a les ser degree. Toxicological data for PCOFs and PCBPs are very limited, as are data on the synergistic effects of two or more of the above by-products. For these conpounds, specific data are lacking on acute lethality, chronic toxicity, carcinogeni city, teratogenic and reproductive effects, mutogenicity, and imnunotoxici ty. There are significant gaps in the relationship between the receptor theory and the induc tion of chi oracne. Oata on such effects as the extent of enzyme induction through specific receptor binding and cell keratlmzation should be acquired through bioas say techniques. HONS 215203 Chemical and Bioassay Analyses Several areas require additional research in order to improve the caoabilities far analysis of PCDOs/PCDFs. Such areas include (1) synthesis and Isolation of addi tional cure PCOO/PCDF isomer standards for use in calibration and methods developnmnt; (2) synthesis of additional compounds which are chemically similar to PCDDs/ PCDFs, and which may represent interferences in PCOD/PCDF analysis {e,,g., diphenyl ethers, hydroxy- and methoxy-QPEs, and PCS*) for methodology evaluation; {31 devel opment of higher resolution GC and/or LC colunrs for Isolation of PCM/PCOF isomers (4) research on more specific detection, quantification, and confirmation nethods. Including possibly positive and negative ion chemical Ionization MS, atmospneric pressure Ionization MS, and MS/MS; (5) research into an isaner-specif1c analytical method to differentiate between toxic and nontoxic Isomers; and (6) research into rapid chemical and/or biological screening nethods; and (7) standardization of sampling methods, such as wipe tests, and establishment of criteria to identify the significance of results obtained via different sampling nethods. Environmental Persistence The environmental pathways of these compounds need to be better defined, neither the pathways nor the rates {other than general overall rates) for their physical mi gratlon and decay have been determined with any certainty. Further research is needed on photolysis to define the role and abundance of hydrogen donors, and the exact conditions under which measurable photolysis will occur in air, water, and soil. The rote of uptake/ metabolism of these compounds by biota and their result ing fate in both water and soil needs further investigation. Oestruction/Remova1 Since these compounds are both persistent in the environment and toxic to animals and humans, more research needs to be directed towerd the developmnt of chemical, pl^yslcal, and/or biological methods for effectively treating wastes containing PCOFs, PCDOs, and PCBPs, as well as soil and weter contaminated with these by products. HOMS 215204 S-1S Section 1 INTRODUCTION background Following promulgation of the Toxic Substances Control Act by the U.S. Congress m 1976, the U.S. EPA Issued restrictions In May 1979 on the manufacture and commercial use of polychlorinated biphenyls (PCBs). Until the above regulations were issued, many transformers and capacitors contained PCBs. It has been estimated that since 1932. 135,000 transformers containing PCBs have been put Into service In the United States The production of PCBs In the United States ceased in 1977. While some PCB equip ment has been removed or replaced, many PCB-contalning transformers and capacitors are still In use. In addition, due to past manufacturing and transformer servicing practices, trace amounts of PCBs have been introduced Into mineral oil-filled equip ment. Hence, It Is likely that, in addition to askarel transformers and capacitors, many of the oil-filled transformers currently in service contain PCBs in concentra tions ranging In the low parts per million. Since 1979, research has shown a potential concern resulting from the presence of same chlorinated aromatic hydrocarbons associated with PCBs, predaemantly poly chlorinated dlbentodloxlns (PCDDs) and polychlorinated dtbenzofurans (PCOFs). Traces of similar compounds, such as polychlorinated blphenylenes (PCBPs), poly chlorinated pyrenes (PCPYs), and polychlorinated chrysenes (PCCYs), have also been reported In seem samples. These compounds occur In extremely low concentrations in PCB liquids. However, same PCDO, PC OF, and PCBP Isomers are acutely toxic as deter mined by laboratory tests on animals and demonstrated In Industrial incidents. Toxlcltles of some of these Isomers are orders of magnitude hi^ier than PCBs. PCDO and PCOF compounds have been found In the original PCB fluids manufactured m the United States, Europe, and Japan. Several PCB accidents In which PCODs and PCOFs were prominent have also been documented. Including the Yusho incident in Japan Involving ingestion of PCOF-contaminated rice oil, and a similar subsequent Incident In Taiwan. More recently, a fire Involving electrical equipment containing MOMS 215205 1-1 PCB fluids in a Bingnamton, New fork, office buildinq produced soot which was #ouna to contain PCOFs and PCODs. These findinas indicate tne possibility tnat CDOs, PCDFs, and otner related cnlortrated aratiatic compounds can fonn ffp* eskarel k'uiqs at nigh temperatures. PROJECT OBJECTIVES The purpose of this study is to determine tne state of knowledge on the formation of PCDOs, PCDFs, and other related cnlonnated aromatics in on-line electrical equip ment containing PCBs, and to provide EPRI with a technical planning tool for formu lating its future research on this subject. The specific objectives of this study are as fo1 lows: e The state of knowledge in the area of PCOOs/PCDFs in utility dielec tric fluids. e Recommend future research needs m the areas of PCDO/PCOF formation in utility electrical equipment, their chemical and biological anal yses, environmental fate, toxicoloqy, and techniques for tneir destruction. Specific information in this report was collected by (1) literature search; (2) attendance at the L84th National Meeting of the American Chemical Society in Kansas City. Missouri (September 1982), and the 3rd International Symposium on Chlorinated Dioxins and Related Compounds in Salzburg, Austria (October 19B2); (3) discussions with leading researcners (worldwide) in the subject area; (4) contacts witn trans former service companies and manufacturers; and (S) discussions with representatives of the utility Industry. REPORT ORGANIZATION Section 2 of this report details a numoer of incidents involving the generation of PCODs and PCOFs. The pnystcal and chemical characteristics of PCODs and PCOFs, along with their formation chemistry and the chemistry of transformers/capacitors, are presented in Section 3. Physical, chemical, and biological degradation of these chemicals and their persistence in the environment are discussed in Sections 4 and 5, respectively. Toxicological effects on laboratory animals and humans are dis cussed in Section 6. The current status of analytical procedures for the determina tion of PCOOs and PCOFs is reviewed in Section 7. This review includes both chemi cal and bioassay analyses. In Section 8, recannenoations are made for future re search needs. These recommendations reflect the identified data qaps m each of the subject areas covered m Sections 3 through 7. Additional information pertinent to the topics discussed in this report is presented in Appendices A through- 0. 1-2 MOWS 215206 Section 2 ACCIDENTS INVOLVING GENERATION OF PCOOs/PCOFs ANO OTHER RELATED CHLORINATED AR01ATIC HVOROCARSONS INTRODUCTION In this section, we examine a number of incidents including fires, industrial acci dents, and contamination incidents resulting from inappropriate waste disposal. The common thread running through these incidents is the production or release of poly chlorinated d1benzo-o_-d1oxins (PCDDs) and dlbenzofurans (PCOFs), some of which are extremely toxic (Figure 2-1). In some of the incidents, there is also evidence of the production of other chlorinated polynuclear aromatics, such as the polychlori nated biphenylenes (PCBPs) and polychlorinated pyrenes (PCPYs) (Figure 2-1). A number of these incidents involve PCB-ftiled electrical eoulpment, including transformers and capacitors. In each case, the electrical eoulpment was caught m an intense fire. The temperatures reached mre extreme, far beyond the temperatures expected during normal (100* to 130`C), or even overload (200* to 250*0, condi tions. Other incidents Include a PCB-f 11 led heat exchanger (in which the PCBs were again heated, probably in an oxygen atmosphere, for an extended period), an indus trial accident involving the menufacturer of trichloroohenol, and an instance of chemical contamination from trichlorophenol manufacturing wastes. ELECTRICAL POWER EQUIP*NT EPISODES Toronto. Canada In December 1977, a transformer substation burned In Toronto, Canada. Very little has been published concerning this incident, but It has been cited that the PCB fluid originally contained 0.05 ppm PCOFs; the ash produced contained 5 ppm (_2). No information was reported regarding distribution, health effects, or chemical analy ses. Norrtalje. Sweden In September 1975, an intense fire burned a capacitor battery north of Norrtalje, Sweden. Eighteen PCB capacitors were burned to various degrees, and same exploded. 2-1 MONS 215207 ter Flgurt 2-1, P*rtlntnt Comical Structures HONS 215208 EkWPnation of the fluid again shooed a large increase in PCOF concentrations (2, 3). Concentrations of PCOFs on nearby objects (fences and pine trees) were below detection limits. Samples of blood taken fran firefighters shooed PCB concentra tions (2-3 to 3.6 ug PCB/g) oithin normal hunts and no detectable PCOF (_2). Bmohariton. New York ~ In February 1981. an intense fire occurred in the utility room of the LB-stbry Smgh amton State Office Building (Binghamton, New York), resulting in the loss and apparent pyrolysis of ISO gallons of PCS fluid from a transformer (4, _5, _6). The fire oas apparently electrical in origin, and produced a fine soot ohich as distri buted throughout the building by the ventilation system (). Analysis of the soot revealed PCBs, PCOFs, PCOOs, and PCBPs. When PCODs and PCOFs were discovered, cleamno was suspended indefinitely, and the building was sealed. Although the building was unoccupied at the time of the fire, over 450 persons claimed that they had been exposed to the chemicals, and many have filed suits against the state (6). No published description of the symptoms experienced by allegedly exposed persons mas given. Stockholm, Sweden In August 19BI, another electrical fire occurred in a 10-kV capacitor battery in a power station in Stockholm, Sweden. Like the Binghamton fire, this incident is thought to nave been caused by a malfunction of the electrical eguionant. Analysis of residues at Stockholm also showed PCOFs and strong evidence of PCBPs. Unlike the Binghamton fire, no PCODs mre found (2). No injury or illness was reported in con nection with this fire. Skovde, Sweden In March 1982, a 400-V capacitor battery serving a high-frequency oven in a metal treataant factory bimed In Skovde, Sweden (2). The capacitors involved were both mineral oil containing and PCB types, and the fire appears to have been unrelated to eguipmnt malfunction. Of 21 PCB-fllled capacitors, 12 were ruptured during the course of the 2-hour fire, which reached at least lOBO'C in some parts of the caoacltor room. The smoke from the fire was distributed throughout the 100 x 200-ft (30 x 60-m) bull ding. HONS 215209 2-3 Analysis of soot simples showed PCDFs, but no PCDDs or PCBPs. Another series of chlorinated compounds was detected; molecular weights and numbers of chlorines cor responded to polychlorinated pyrenes (PCPYs), but no confirmation or quantitation was possible due to the lack of standards H) NONELECTRICAL industrial episodes _ Chick Edema A. L. Young has reviewed the dioxin poisoning episodes known collectively as "chick edema" (_7). During 1957, millions of chickens died after consuming feeds contami nated with l,2,3,7,8,9-hexach1orodibenzo--d1oxin (HCOO). Clinical signs of the disease included labored breathing, unsteady gait, subcutaneous edema, and sudden death. The birds were found to have excessive fluid m the abdominal cavity and the pericardial sac, and swollen, palid livers. Mortality was reported to be as high as 50 percent. The contaminant, known as "chick edema factor," was isolated in the early 1960's, but the structure was not elucidated until 1967. Chick edema disease was essentially dormant through the 1960's until another out* break in 1969. On this occasion, it was learned that dioxins had accumulated in the edible tissues of the chickens. In each case, feeds had been supplemented with fats and fatty acids, generally by-products of stearic and oleic add manufacture. It was speculated that the fats were contaminated with commercial chlorophenols, which were eontaannated by or converted to dioxins during heating. fusho (Japan) In October 1968, Japanese health officials became aware of an epidemic disease that came to be known as "Yiisho." Victims exhibited such symptoms as acne-1 ike skin eruptions, pigmentation of the skin and mucous membranes, swelling of the eyelids, increased eye discharge, hyperkeratotic plaque on the soles of the feet and palms of the hands, and hyperemia of the conjunctiva (B, ). The cause of the disease was eventually traced to certain lots of rice oil (Kanemi) produced during February and Mareh 1968 () It is speculated that the oil was con taminated during vaeuum distillation via pinhole leaks in the piping of a heat ex changer. The heat exchanger used PCB fluid as the heat exchange medium, and it maintained the distillation tank at 230*C. GC analyses showed 2000 ppm "Kanechlor" brand PCB fluid In the oil (9). HONS 215210 2-4 A 1971 report set the number of Yusho cases at 1001 (). A subsequent report set the number at 1200 (.10). Patients shoimd detectable concentrations of PCBs in their subcutaneous fat. Birth defects and retardation of normal growth were also noted Of 13 babies born to women affected with Yusho, 2 were stillborn, IQ had hyperoiomentation, and 9 had increased eye discharge IB). Pigiwntation disappeared m 2 to 5 months (^). Children initially exhibiting retarded growth caught up with a nor mal control group within a few years (12). After the general potential for PCOF and PCOD contamination of PCBs was reported, used PCB fluid and the rice oil Involved in the Yusho ooisoning were reanalyzed. No significant levels of PCOD or PCBP were found (.10.). PCOF was found in both fluids (15 6 and 5-65 ug/g, respectively) (.3. .10* .13, .1*, 15). Other analyses of the nee oil indicated that, in addition to the PCOFs present, there were also other chlorinated compounds, including quaterphenyls (PCDs), ouaterphenyl ethers (PCQEs), and terphenyls (PCTs) (Figure 2-1). Most of the "unknown organo-chlonne compounds" were identified as polychlorinated quaterphenyls (PCOs), and most of these were hepta- or octachloroqueterphenyls (16). Yusho (Taiwan) Health authorities in Taiwan reported a second outbreak of Yusho in the Taichung and Chanonua areas of Taiwan in 1979 ( 17, 18). It is reported that 1900 people were af fected by PCB-contaminated nee oil, but no further information was oiven concerning the oriqin or production of the oil (.17.). The researchers presume that the oil was contaminated by the heat exchanger durina manufacture (.18.). Analysis of the contam inated oil showed both PCBs and PCOFs Q8). Blood samples from affected patients also showed PCBs (.17) and PCOFs (.19). St. Louis, Missouri Between February and October 1971, trichloroohenol (TCP) distillate residues fron the Northeastern Pharmaceutical and Chemical Company (NEPACCO) in southern Missouri were collected by a salvage oil company in St, Louis (Figure 2-1). The 13 , 000 gal lons (68,130 liters) collected were dumped into a large holding tank, along with other waste oil and sludge. During May and June of that year, the satvaqe company sprayed three horse arenas with waste oil from the tank to control dust. In subse quent weeks, birds, mice, cats, dogs, and horses exposed to these areas became sick and died. Among surviving horses that were pregnant, 26 abortions occurred, and many foals died soon after birth, soem with manifest deformities (20, .2U- Several 2-5 HONS 215211 children exposed to the contaminated arenas became ill, with symptoms including chloracne, headaches, aching joints, and inflammation of the kidneys (21), One little girl develooed hemorrhagic cystitis following repeated exposure to one of the arenas. In 1977, Beale, et a 1., reexamined her and found that she had grown normally. Detailed physical, chemical, and neurological examinations of the child and her mother and sister (who *ere also exposed, but to a lesser extent) were nor mal (_7) Laboratory analyses of soil fran one of these arenas showed approximately 5000 opm TCP, 30 ppm TCOO, and 1350 to 1590 ppm PCB (21.). 2,3,7,B-TCDD was found to be a major canponent of the PCODs, and 1,3, 7, S--TCOO a minor one; no tn- or penta-CODs were detected (C. Rappe, Persona) Communication), [n addition to PCOOs, approxi mately 3 ppm PCOFs were found (mainly 2.3,6,8-TCOF, with some 2,3,7,8-TCOF and traces of penta* and tri-CDFs) (C. Rappe, Personal Communication). Furthermore, polychioroxanthenes (PCXs) (Figure 2-1) were found to be a major component (primar ily 1,2,4,6,9,9-hexachloroxanthcne, "1th traces of pentachloroxanthene) (C. Rappe, Personal Communication). It should be noted that hexachloroxanthene is a major lagiurity in commercial hexachlorophenes (C. Rappe, Personal Communication). The pharmaceutical company waste storage tank, left undisturbed, was analyzed in 1974, and showed approximately 300 ppm TCOOs, 30 percent TCP, and no PCBs (.21)- The PCBs in the soli apparently came from the wastes of another, unidentified canpany. Following an anonymous phone call in 1979, the CPA began further investloations of NEPACCO's waste disposal practices. Eventually, 30 sites in the Spnnq River Basin of southwest Missouri were identified. Results indicated widespread distribution of 2,3,7,8-TCOD attributable to NEPACCO's waste disposal (22). Save so, Italy One of the largest accidental TCOO Incidents occurred In a TCP manufacturing plant north of Seveso, Italy, In 1976. The reaction went out of control, and the vessel vented to the atmosphere, releasing a caustic cloud which settled on several com munities to the south of the plant. Because the exact nature of the chemical cloud was not known, the company tried to deemphaslze the levels of toxic chemicals in volved, and no immediate public health precautions were taken (, 23, 24). Within a week of the accident, widespread phytotoxicity (poisonous to plants) was evident, small animals were dying, and children were being admitted to hospitals HONS 215212 2-6 w,th burns. Tests confirmed thet the chemical cloud consisted primarily of 2.4,5. trichloroohenol, containneted with 2,3,7,8-TCDD end traces of 1,3,7,8-TCDD. 2,7- or 2 g-di-CDD. 2,3.7-tri-COO, and mono-CDO (25). Minor amounts of tri- and tetra-C3F acre also found in the first samples (25_). The last sample was analyzed m Marcn 1977 for the presence of other neutral and phenolic compounds. Apart from the PCDOs nmntiofied aPove. traces of di- and tnchlorophenoIs. tet-a- and heiachlorooeniene. PC9, and DDE were detected. Areas where TCDO concentrations measured 0.001 ppm or more were evacuated (_7_) Children in other local areas were evacuated during the day; such areas were closed to nonresidents, and veoetable growino and animal breeding were forbidden (7, 23). Analyses indicated that 98 percent of the TCDO was sorbed in tne too 4 cm of the soil (7.)- Estimates of the total TCDO deposited varied from 450 to 3000 g (_7, .231. Oecontamnation measures that were eventually taken included vacuuming, scrapma. and washing of waits, floors, and ceilings; removal of most of the furniture; and removal of the top layer of soil in gardens and orchards. Reports on health Impacts resulting from the accident vary concerning the nui*er of people affected. The number of chtoracne cases ranged between 134 and 187, with most of the cases occurring in children (7, 26). Chtoracne symptoms were rarely severe and seldom recurred. Where they recurred, ft was usually in mild to very mild cases (). Neurological damage wts reported in 33 adults, but no liver or gas trointestinal damage was documented (27). Another report indicated no abnormal neu rological findings, and no laboratory evidence of significant fiepatotoxici ty or de ranged porphyrin metabolism (7.). Analyses througn 1980 indicated no significant changes In either the general or cause-specific mortality rate m the area (28). FORMATION OF PCOO, PCOF, ANO OTHER CHLORINATED ORGANICS PCOQ, PCOF, and other chlorinated organics have been formed m the aftermath of these accidents, and have been detected both alone and m various comoinations. Table 2-1 simnerites the toxic products formed in each case. It is difficult to comare the results from each incident because of the different sampling methods and sample matrices (wipe tests, soot seeples, etc.). PCOO Incidents PCODs have been accidentally produced in the course of a few of these incidents, and under widely varying circumstances. The PCOOs (or simply "dioxins") found in the 2-7 HONS 215213 wake of the explosion in Seveso have become a byword for industrial environmental contamination. As much as 5.5 to 6.5 tb (2.5 to 3 kg) TCOD vere released (_7), al though some estimates are much lower A$ oreviously stated, 93 percent of the TCDD is strongly sorbed in the top 1.5 in (4 cm) of soil. Ongoing decontamination efforts have been focused on removing topsoil, depositing it in the evacuated tone, and finding a way to decontaminate it, possibly through extended.biological degra dation (7). Tabte 2-1 SUMMARY OF CHLORINATED AROMATIC HYDROCARBONS OETECTEO 0IKING DIFFERENT ACCIOENTS *CCl4>nt TgfORtO scoot scirt scot sCOEt 211 sCISt CPTl seen sen scoot t toftrtftce sw 2 Wrrtaljt (1*1(1 tl-107 i IliWiwU" 2.1It Stoctfcs (IMt) Stw* u*MIJ MW tl-10* V*/* 0.01 0.171 e 4, 2 10 Chlct EXtwi WitWo. JawM *u*n, St lav it Sent* I'Ll wm 0 11 1M MW 10 MW 0^1 * * * 7 it m 21 31 OwmiMcittew ww i MW MtlWk XM to lack of ttMXXMt Seveso Is the only TCOO Incident for which "operating conditions" are knowi. Ther mographic records at the plant showed an uncontrolled Increase in the temperature of the reaction vessel, following failure of a safety shutdown device. The mixture was heated at 230 to 240*C for 4 to 5 hours and 3 to 4 atm before a safety disc rup tured, releasing nearly the entire mixture (7j, HONS 2-8 The waste oil, which contained still bottom residues from TCP manufacture and was used to control dust in Missouri horse arenas, contained sufficient TCOO to yield 30 ppm TCOD in a soil sample (2). When the waste oil storage tar* was examined later, it was found to contain 300 ppm TCOO (21). As noted above, 2,3,7,8-TCOO has been found m disposal sites throughout the Spring River Basin. As of mid-1982, 30 pos sible sites had been identiflew, and investigations had accounted for approximately 110 lb of still bottom-derived TCOO. In the Binghamton State Office Buildlno fire, PCOOs imre formed during the combus tion of transformer fluid consisting of PCBs and chlorinated benzenes. The hew rork State Department of Health analyses found 2.8 and 2.g ppm (uq/g) 2,3,7,8-TCOO in the soot when duplicate analyses were run on a single sample (). Another s*nole of soot was analyzed and reported to contain 20 ppm PCOO and 0.6 ppm 2,3,7,8-TCOO (291. PCOOs i*re not found m the residues of the other transformer fires. Rappe, et at., attributes this difference to the presence of chlorinated benzenes in the dielectric fluid, as PCOO Is known to be formed In the pyrolysis of chlorinated benzenes (3). PCOF Incidents PCOFs have been detected m more incidents than PCOOs; they have been found m all PCS fires in which measurenmnts <*re taken, as well as in the Yusho poisoning in. cldent. In the Binghamton transformer fire, 273 and 124 ppm PCOF were reported for duplicate analyses of a soot sample (). It Is thought that this represents 2,3, 7.B-TC0F, but positive confirmation of the structure was not possible due to a lack of Internal standards. Rappe reported additional analyses of soot, giving 12 ppm for 2,3,7.B-TC0F, 28 ppm for total tetra-COF, 670 ppm for penta-COF, 965 pom for nexa-COF. 460 ppm for hepta-COF, and 40 ppm for octa-COF (29). PCOFs were observed in the residues froe all the fires involving PCB-f 11 led caoacitors. At the transformer station in Norrtalje (in 1978), PCOF concentrations ranged frm 1.1 p|B in an undeemged capacitor to 75 ppm in the fluid left inside an ex ploded capacitor, and 52 ppm on the outside of that same exploded capacitor (2). ho PCOFs were detected on nearby trees, or in the blood samples taken from firefighters (1). Absolute confimmtion of specific Isoamrs was not possible (2,. _3). In the 1962 fire In Skovde which burned 21 PCB capacitors, PCOFs were found on the floor in and near the capacitor battery, on the wall of the capacitor room, and on a bench; 2,3,7,8-TCOF was found In all but the last Instance (). In the 1982 Stock holm fire, seven tetra-COFs and 11 penta-COFs <mre identified and quantified. HONS 215215 2-9 In the Yusho incident, PCDFs were found in the rice oil. The oil showed concentre* tions of 4 to 5 op* ifter contamination with approxmutely 1000 OP* PCS. Tests of venous PCS mixtures indicate PCOF contamination of 1 to 18 pom; using 10 pom PCOF in PCB and 1000 ppm PC8 in rice oil, 0.01 ppm PCOF in rice oi I would be expected. Actual findings of 5 ppm support the view that PCOF was formed fron the PCS after prolonged heating (U_, _15). In a laboratory simulation of the steam distillation process used, PCOFs were formed fron the PCSs (_32_). " When Yus ho broke out again in Taiwan, researchers reported S3 to 405 ppm PCB in the toxic oil and 0.18 to 1.68 ppm PCOF (_18J- Fron blood sample analytes, it also ap* peered that soam oils, which re not available for analysis, had contained much higher levels of PCB (_18_). No identification of the PCOFs was reported. Other Polychlorinated Oroamcs In addition to PCDOs and PCDFs, a number of other polychlorinated oroamcs nave been detected In samples from these incidents. PCQs, PCQEs, and PCTs ire all detected in Yusho oil froe Japan (_16); PCQs and PCTs were detected In Yusho oil in Taiwan (18). PCBPs were detected in the residues of the Binghamton and Stockholm (1981) fires U, ). Evidence of polychlorinated pyrenes (PCPYs) and polychlorinated chrysenes (PCCYs) was found the 1982 Stockholm fire and In the Skovde fire; poly* chloroxanthencs (PCXs) were found In the St. Louis incident. HONS 215216 Section 3 PCDD/PCOF/PCBP FORMATION physical AND CHEMICAL CHARACTERISTICS OF PCODs AND PCOFs Both the dlbento-o-dl oxi n (or dioxin) and dlbenzofuran compounds have a triple-ring structure consisting of two benzene rings connected by a third ring, [n the dioxin (shown below), the connection is through a pair of oxygen atom. * The middle ring of the dlbenzofuran contains only one oxygen atow, as shown below. Both compounds can have chlorine atom at any or all of the numbered positions on the benzene rings. Theoretically, there are 75 possible different positional iso mers of chlorinated dioxins, frae mono- to octechloro, and 135 different chlorinated dlbenzofuran isomers (Table 3-1 and Appendix A) (33, 34). Very little published Information is available on the chemical and physical proper ties of moat of these compounds, particularly the polychlorinated dlbenzofurans (PCOFs). The unsubstituted dioxin moiety is stable to at least 700*C and possibly to U50*C (33). The 2,3,7,8-tetrachloro isoamr is the most studied isoamr due to its toxicity. This Isomer is symmetrical across two planar axes. It Is sparingly soluble in water and In most organic solvents; it Is relatively inert to acids, lases, oxidation, and reduction, and Is extremely lipophilic. In general, chemical stability and llpaphlllclty increase with increasing halogen content, although higher chlorinated isomers are more readily subject to attack by strong acids and MOWS 215217 3-1 bases. 2,3,7,8-TCDO Is a solid at roan temperature, melting at 305*C- The octachloro Isomer melts at 130*C. Table 3-1 ISOMERS OF PCOFs AND PCODs Chlorine Substitution Mono01TrlTetraPentaHexaHeptaOcta- Number of Isomers PCOFs PCDOs 42 16 10 28 14 38 22 28 14 16 10 42 11 135 75 CHEMISTRY OF FORMATION OF PCODs ANO PCOFs A suemary of chemical Pathways to PCDOs, PCOFs, and PC8Ps is given in Figure 3-1. Polychlorinated Dlbenzo-p-QloxIns (PCDOs) There are a limited number of direct precursors and reactions by which PCDOs can oe formed. In order to be a dioxin precursor, a compound mist meet two conditions (33): a The coeound mist be an ortho-substituted benzene ring in which one of the substituents Includes an oxygen atom directly attached to the ring. e tt must be possible for the two substituents, excluding the oxygen, to react with each other to form an independent compound. An example of the ideal dioxin precursor is as follows: : MOWS 215210 3-2 OH Ir Figup* 3-1. Chwlctl Patlw*ys to PCOOs. PCDFs. *nd PC BP* MOHS 215219 3-3 A reaction between two of these molecules could yield a dioxin and l XY molecules, as follows: Although the number of direct dioxin precursors is relatively small because of the conditions set forth above, there are many indirect precursors, i.e., compounds which do not fit the model, but which can degrade or be transforwd into direct pre cursors. This wi 11 be discussed in more detail later Buser (5_) and Rappe, et al. (_35_). have identified the following basic reaction for chlorinated dioxin formation: Dimerization of chiorophenols and phenoxy acids (pyrolytic and ohotochemical) - A bimolecular reaction in which the relative retio of isomers formed depends on the concentration of the chiorophenates; yields decrease in more dilute systems, such as contaminated mineral o! Is. a CycIllation of predioxins (pyrolytic and photochemical) - Unimolecular, and thus concentrationsndependent. a Dechlorination of higher chlorinated PCDOs - Unimolecular, and thus concentration-independent. a Direct chlorination of dibenio-o^dioxins. The dimerization reaction (rxn 1) basically follows a two-step nechamsm (33) (rxn 2) The intermediate diphenyl ether compound is called a predioxin. Typically, a predi oxln will either cycllze to dioxin or break down to the precursors. For example, i one test, Irgasan DP30D, a compound with a predioxin configuration HONS 215220 3-4 *11 heated to 980*C (36) The reaction yielded both dioxins and precursors. Be cause of such competing reactions, dioxins are seldom found in more than trace quan tities. It has been shown that the higher chlorinated predtoxins are amenable to self-condensation, while the lower chlorinated predioxins apparently do not readily cycllie further (J7). For instance, in one test, irradiation of 2,4-dichlorophenol yielded a predioxin, but no further reaction (33). Temperatures for dioxin formation by dlmenzatlon/cyclization have bean reported at values from 180*C to 620*C (33). Sandermann, et al. (cited by Choudhry and Hutzinger) (_30_), obtained octachlorodtoxtn (XDD) (6 percent) and hexachlorobenzene (51 percent) upon heating 100 g of pentachlorophenol m a closed tube at 300*C. Espo sito, et al. (33). reported quantitative yields of OCDD fron heating the sodium salt of pentachlorophenol at 360*C. Ahllng and Lindskog (_39) found that combustion of tn- and tetrechlorophenols at 500* to 600*C can yield PCDD. Duckett (40) reported temperatures for dioxin formation from pentachlorophenol ranging from 310* to 370*C. However, the formation of dioxins froa chlorophenols Is exothermic, although there are no published data on the amount of heat released (41, 42). This fact, combined with the need for heating to initiate the reaction, suggests that one or more of the reaction steps In the dioxtn formation sequence has a high activation energy. The high activation energy contributes directly to the low yields. In addition, the boiling points of many dioxin precursors are lottr than the reaction temperature range. Thus, there is a loss of precursor molecules before reactions can occur (40). Host laboratory syntheses of dioxins rely on closed reaction vessels or pres sure to keep precursors in the liquid state. Another major source of penta- and tetra-COD 1 sonars Is photochemical dechlorination of higher isomers, mainly 0C00 (43). The principal toxic isomers, 1,2,3,7,8-pentachloro and 2.3.7,8-tttrachloro. are not as readily fommd as other penta- and tetrachloro isomers. The major hapta-CDD fornmd is the 1,2,3,4,6,7,9-substltuted isoamr. Indicating a preferential loss of lateral (2,3,7-, or 8-) chlorine atoms (44, 45). The major hexa-CDO formed is either 1,2,4,6,7,9- or 1,2,4,6,8,9-hexa-CDD (44). The major penta-COO is expected to be the 1,2,4,6,9-substituted isomer, and the major tetra-CDD is 1,4,6,9-tetra-CDD (44). These reaction products show a preferential removal of the lateral chlorine atoms. Lower Isomers (di- and trlchloro) may be formad by irradiation, but they decopose via the same route, and thus do not acemulate (46). HONS 215221 3-5 Othtr know* sources include condensation of catechols wi tn polychlorobenzenes or chloronitrobenzenes (22) {figure 2-1). Pyrolysis (S20*C) of chlorobenzenes in me presence of air yields PCOOs (_33. 2' Since chi orobenzenes do not fit the csndi. tions set down earlier for dioxin precursors, 3user (25_. 9_) has oostulated that the alkaline hydrolysis of chlorobenzene to PCOO involves a two-stee conoensation pro cess from ortho-chlorinated pnenoxy anions via a polychlorinated "2-phenoxy pnenate. This dimerization is exotnermic. Below approximately 180*C, usually very minor amounts of PCOOs are formed {<1 ppm), but significant quantities (1 to 10 oercent yields) result at temperatures over 180*C (25). Dioxins have also been detected in residues from a variety of combustion processes, including municipal incineration, the burning of fossil fuels, wood in wood stoves, and cigarettes (SO). Precursors of dibenzo-o-dloxins are also known to be formed after simply adding hyorogen chloride to the flam of a Bunsen burner (20). Most Investigations have been centered on municipal incinerators and industrial heating plants. Olie, et al. (SO), for example, analyzed fly ash samples from a municipal incinerator in Netherlands, and found average PCOO concentrations of 2056 ng/g. Heiachloro isoamrs predominated, followed by heptachloro isorwrs. Ltbertt, et al. (52), found total concentrations of PCOOs and PCOFs on the order of 1300 cob in Italian municipal incinerator fly ash. They suggested phenolics plus chlorine or HC1 as precursors. Buser, et al. (52). also detected PCOOs m Swiss imjntooal fly ash. The major isoamrs detected were the same as those formed in the pyrolytic dimerization of the more common chiorophenates, namely the 2,4-dt-, 2,4,6-tn-, 2,3,4,6-tetra-, and pentachlorophenates. The mechanism(s) of PCOO formation in fly ash is presently unknown. Two suggested sources are: a Condensation of chlorophenols. a Thermal synthesis from any organic materials and inorganic chloride (the "trace chemistries of fire" hypothesis) (54). HoMver, even Ideal PCOO precursors, such as polychlorophenols, show very low PCOO conversion efficiencies. Thus, although every chlorinated organic compound or HONS 215222 3-6 orgin'C caepound/lnorganlc chlorine mi* might be regarded as a potential precursor, the r(n^ of likely precursors for formation of detectable quantities of PCDDs is probably not thit &rAd (.SSh Consequently, it is unlikely that organic material/ inorganic chloride reactions will produce PCOOs. The nigh-temperature am oxidation of coal in the presence of chlorine, MCI, or <aaC 1 has been shown to yield PCODs (47) However, coal is a commercial soiree of benzene and cumene {a phenol raw material. Figure 2*1), so that formation of Chlorinated benzenes and phenols is a probable intermediate step. Thermal decomposition of lig nin (Figure 2-1) and polyvinyl chloride (typical constituents of municipal refuse) can yield phenols, cresols (Figure 2-1), catechols, benzene, chlorobenzenes, HC1, etc. (46. 56). Thus, rather than just any organic material serving as a potential precursor, it seems more likely that those compounds which can be pyrolyzed to ben zene, chlorobenzene, phenols, catechols, and/or inorganic chlorine are more prooaole indirect precursors. The *chan1sm(s) by which PCDDs are formed during combustion is still not well understood; thus, the effects of combustion temperature, fuel characteristics, catalysis, and other parameters have not been determined (7). Townsend (5) has suggested that dioxins created within a combustion source probably have a compo sition which reflects the chemical conditions within the source. Upon emission, a chemical equilibrium process takes place, yielding isomer ratios reflecting the chemical conditions of the atmosphere. However, this theory has not been generally accepted. There has been some question regard!nq the role of PCBs in PCDD formation. In 1981, an electrical fire In the State Office Building in Binghamton, New York, resulted in the pyrolysis of PCS fluid froa a transformer (, 6K Soot froe this fire con tained PCDFs and PCDDs. PCBs do not fit the precursor model discussed earlier, and PCDDs would not be expected to form readily in a mixture of biphenyl compounds (59). Conceivably, PCBs could be oxidized to polychlorinated hydroxybiphenyls which could, in turn, cycllze to form dioxins (60). Another reaction that has been suggested is (47); ei* ei ci.., However, neither mechanism quite fits the thermal decomposition pattern .for PCBs. 3-7 MONS 215223 The PC6 fluid at Binghamton was * mixture of 65 percent Aroclor 1254 end 35 percent chlorobenzenes. Rappe, et el. have suggested that pyrolysis of the cnloroben. zenes is a more likely source of the PCOOs than PC8s. This reaction was discussed earlier. Polychlorinated Qlbenzofurans (PCOFs) ~ While PCOFs and PCOOs can share son of the same precursors, PCBs are the aost com* mon source of PCOFs. Buser and Rappe (3) have identified four basic nchanlsms of PCOF formation froa PCBs: *ci. {rxn 5) ci. } * wei ci, ei, cf. rxn 6) W ci ci T C|i o ct (rxn 7) * 01, 01, 'O' '01, (rxn 8) Reaction 4 involves the loss of the ortho-Cl. Reaction 5 Is a loss of HC1 involving a 2,3-chlorine shift. Reaction 6 has a loss of ortho-HCl. Reaction 7 Involves a loss of ortho-h. PCOFs have been identified as trace contaminants at the ppm level in a nuirter of American, European, and Japanese canardal PCB mixtures (.13, 3., 62., 63). HONS 215224 3-8 Concentrations and isomer ratios vary *fth the type of PCB and the country of ori* gin, probably ** * result of differences in manufacturing conditions (Table 3*2). For instance, Bowes, et al. (83), detected PCOFs in American-manufactured Aroclor 1260 and 12*8 at total concentrations of 0.8 and 2.0 ppm, respectively; none re detected m a single sample of Aroclor 1016. The total quantities of PCOFs in German Clophen A-60 and French Phenoclor OP-6 were estimated to be 8.4 and 13-6 oom, respectively (62). PCOFs have also been detected m several Japanese-made PC3s at concentrations frae 1 to 20 ppm, with the highest concentration In a sample of KaneC hi Or AC-400 Ul, 14). Table 3-2 LEVELS (ug/g) OF PCOFs IN C9VCRCIAL PCBs Sample Aroclor 1248 (1969) Aroclor 1242 Aroclor 1242 Aroclor 1243 Aroclor 1254 (1969) Aroclor 1254 (1970) Aroclor 1254 Aroclor 1254 (KK 602) Aroclor 1254 Aroclor 1260 Aroclor 1260 (1969) Aroclor 1260 Aroclor 1260 (AK 3) Aroclor 1016 (1972) Clophen ABO Clophen T64 Rienoclor OP-6 Prodelec 3010 Kanechlor 400 Mitsubishi (used) Trl- 0.1 0.06 0.1 0.41 2.13 Tetra- Penta- Hexa- Hepta- Total 0.5 0.07 2.3 0.25 0.1 0.2 0.02 0.05 0.1 0.3 0.1 O.B 0.2 R.O. 1.4 0.3 0.7 1.08** * 4.00 1.2 0.03 2.2 0.7 0.2 0.4 0.2 0.1 3.6 1.0 0.4 0.9 0.3 R.O. 5.0 1.73 10.0 0.35 3.30 0.3 0.003 R.O.* 0.81 1.4 0.9 0.4-0.6 0.02 1.9 1.10 0.5 0.5 0.3 R.O, 2.2 2.45 2.9 0.07 * 0.53 - 2.0 - 0.15 - 4.5 - 1.9 - 1.7 - 1.5 * O.B - 0.2 - 5.6 1.35 - 3.B 1.0 2.2 - O.B -- 0.B2 - 8.4 5.4 13.6 2.0 - ca. 20.0 10.0 Reference 63 65 15 64 63 63 66 64 15 64 63 15 63 63 63 64 63 64 66 64 N.D. * None Detected. * Major Isoamr 2,3,7,8-tetra-COF. PCOFs can also form during PCB use. The best known example of this Is the "Vusho" Incident In Japan, which Is dlseussed in npre detail In Section 2. The PCBs, *iich MOWS 215225 3-9 had leaked frae a heat exchanger, had PC0F/PC8 ratios 200 to 250 times higher than that of the original Kanechlor KC-400 (H.* 64). Furthermore, the PCOFs in the usho oil contained the sane isomers as a sample of used PCBs U). This suggested that most of the PCOFs had been formed in service (62_). Similar PCDF isonr ratios have been detected m other used Japanese PCBs (41). In an experiment designed to duplicate the Yusho oil incident, Morita, et af. (67), heated PCBs in air for 1 Meek. PCOFs Mere formed at temperatures over 270*C, reach ing a maximum level at 300*C, and then declining at 330*C. suggesting decomposition primarily, di - and tnchloro dlbenzofurans Mere formed m this experiment, while tetra- and pentachloro isomers were found in the Yusho oil. Jansson and Sundstrom (2_) showed that, at 550* to 650*C, 0.5 to 2.8 percent of PCBs converted to PCOF, Mhile no net formation was observed above 700*C. Other sources of PCOF include condensation of chlorophenates and chlorobenzenes, photochemical degradation of hl^ier chlorinated isomers, cjcllzation of polychlori nated diphenyl ethers and pol chlorobenzenes, and direct chlorination of dibenzofurans (45). Buser (*5_) showed that octachloro isoeers could be photochemical 1y dechlorlnated to tetra- and oenta-COFs. The preferential positions for photochemi cal loss of chlorine In the dlbenzofuran system have not been studied. In their discussion, Choudhry, et al. (47). noted that both chlorinated diphenylethers and chlorobenzenes wtll yield PCOFs and PCDOs when heated to 600* to 620*C. Chlorophenols and chlorobenzenes can form PCOFs via an intermediate polychlorinated diphenylether (PCOPE) (25, 33). WWW Cl * * WjO PCOPEs usually occur as contaminants in chlorophenols, and are known to form PCOFs by thermal, photochemical, or metal-catalyzed reaction (68). PCOPCs and PCOFs probably form early In the reaction when large concentrations of chlorobenzenes are still present, whereas PCOOs are formed toward the end of the react 1 on (68). Fires In electrical equipment containing PCBs have resulted in PCDF formation. Jansson and Sundstrom (_2) Investigated the aftermath of a 1976 capacitor fire in Sweden. Whereas the original PCB contained 1.1 to 1.3 ug PCDF/g (amounts of mono-. HONS 215226 3-10 di-, tri-, and tetri- isomers about eoual), PCB samples from inside the exploaeo capacitors avenged 81 ug PCDF/g PC8 (approximately one-half of PCOF di- itowr, one-third mono-, and the rest tri-, with a trace of tetn-}, and samples from the inside surfaces contained 45 to 107 ug PCOF/g PCS (predominantly di- and mono- 'so bers mth some tri- and a trace of tetri-). Soot from a lg77 transformer fire m Toronto contained 5 ug PCC'Vg, while the original PCS contained OTOS ug PCDF/g (.2). During the course of the 1981 Binghamton State Office Building electrical fire, the secondary Bushing of a 1100-gal ton (4160-liter) transformer cracked, releasing about 180 to 200 ( 680 to 760 liters) gallons of transformer fluid (65 percent Arcelor 1254 PCB, and 35 percent tri - and tetrachlorobenzene) (69., 70). Chemical-contaminated soot was spread throughout the building by the ventilation systme. Two soot compo site samples collected from the stairwells contained 120 and 270 ppm 2,3,7,8-TCDF (70). Analyses of soot samples by Stalling and Rappe/Buser showed PCDF/2,3,7,8-TCOF levels of 756 ppm/3.7 ppm and 2160 ppm/ 12 ppm, respectively (.70). In September 1982, a 500-unit capacitor battery at a steel mill in Surahammar, Swe den, was ignited by 10 tons (9090 kg) of molten steel. The capacitors were filled with PCSs (2 tons [1818 kg]) and mineral oil (3 tons [2727 kg]). The fire burned for 2 to 3 hours, and filled the entire building with smoke. Table 3-3 presents the results of analyses of wipe samples. There have oeen several other capacitor fires after which PCOFs were found, such as those at a paper mill in [metre, Finland (August 1982), and Volvo in Skovde, Sweden. Table 3-4 presents a comparison of the gross PCOF from several incidents. Figure 3 2 shows very similar TCDF isoernr distributions from these same incidents. PCDFs have also been detected in fly ash samples from municipal incinerators and in dustrial heating facilities. Incinerator fly ash from the Netherlands averaged 1309 ng PCOF/g (51). Incinerator fly ash from Italy averaged about 1300 ppb of cambtned PCDO and PCOF (52), Fly ash from an industrial heating facility and municipal in cinerator in Switzerland contained 0.3 and 0.1 ppm PCOF, respectively (S3). Thus far, the exact source of the PCDFs in fly ash is unknown. Buser, et al. (53). pyolyied commercial PCSs in quartz tubes, and produced the same relative PCOF isomer ratios that mire detected in the Swiss fly asn, suggesting that PCBs were tne source of the PCOFs. Other researcners have suggested that the similar isomer distribution is a function of the reaction surface (quartz tubes are SIOj, and fly ash is gener ally over 50 percent Si0^), and do not necessarily indicate the same precursors (72). NONS 215227 3-11 Table 3-3 PCOFs found AT SLRAHAMMAR, SWEDEN (ng/m2)* Samole Location Capacitor Room (Sample 1) Capacitor Room (Sample 2) N.E. Corner, 10 m height S.E. Corner, 10 m neight N.E. Corner, floor S.E. Corner, floor 10 m Outside, downwind 300 Outside, downwind After Cleaning (Samole 1) After Cleaning (Sample 2) TCQF 4000 1100 1250 480 100 90 <250 <250 <20 <40 2.3. 7.8 TCDF Cl* Cl* ci* 875 3300 1800 1500 365 1250 940 625 300 355 150 65 120 210 140 60 25 27 15 5 22 25 17 17,. <25 <25 (5) <60 (10) SB * <25 <25 (S) <60 (10) <30 <4 <10 <12 <15 <8 <20 <20 <30 oa 300 145 13 30 2 a 17' <12 2! 6 Source: C. Repot, Personal Communication. * Wipe tests. ** Possibly due to Cl^-Cl^j PC8 in casting sand. Table 3*4 PCDF LEVELS IN PCB FIRES Site Skotvde (foundry, well-ventilated) Skoevde (basement capacitor rooe) Dennllcen, Sweden Surahammer, Sweden Imatra, Finland [metre, Finland Binghamton, New fork PCOF Levels 0.01 ug/m2 0.8 ug/*2 1.5 ug/m2 1-10 ug/m2 1-3 ug/m2 1-5 ppm 2160 ppm PCBP Levels None None 1.5 uq/m2 Less than PCOFs Less than PCOFs Less than PCOFs * Quantification not possible due to the lack of standards. 3-12 HONS 215228 Binghamton, Nm York Oannlkan, Stockholm, Sweden Volvo, SkSvd, Sweden Surahammar, Sweden Imatra, Finland TCOF Standard wixture Figure 3-2. TCOF Isomer Distributions at F1v PCS Fires 3-13 HOWS 215229 TRANSFORMER/CAPACITOR CHEMISTRY Askarel transformers are those transformers designed and manufactured to contain a liquid fire-resistant, electrically insulating coolant (askarel), consisting of 60 percent (600,000 ppm) to 100 percent (1,000.000 ppm) comwrcial PCS products ; 56), and uo to *0 percent tr ichlorobenzene (U. The term "PCB transformer," as oefmeo in 40 CFR, Sec. 761.3(y), includes all askarel transformers (as defined aDOve), plus transforms filled with mineral oil but contaminated with PCBs at concentrations of 500 ppm or greater, 'Most PCB-fi 1 led electrical equipment is desmned to operate at temperatures of 100' to 130'C, and transformer "hpt spots- are typically no greater than 135*C (6) All units may contain air, but most newer ones have an inert atmosohere, usually nitro gen. Askarel units are sealed, but some seals can deteriorate with time, oermittino the entry of air and water. Hydrochloric acid can be oenerated, lead mo to interna) corrosion. Water can cause arcing. Internal transformer malfunctions can cause bushings and winding connections to becone white hot. During these occasional short-circuit episodes, temperatures reach 200' to 2$0'C for a few minutes. In cases of an arc where the transformer falls, maximum temperatures may reach 1000'C; these failures occur within fractions of a second (6). Capacitors typically oper ate below 100'C. In those capacitor failures resulting In rupture, heating and in. ternal arcing may have occurred prior to rupture. Samples df used oil taken from two capacitors (Prodelec 3010) have been analyzed for PCOFs; no apparent Increase was detected (see Table 3-2) (_I9.. 6). The formation of PCOFs when capacitor oils are exposed to sufficient oxygen and high temperatures was investigated by conducting laboratory tests with Prodelec 3010 (_19). The data indi cated that both sparking and soldering did not result in an increase in the concen. tratlon of PCOFs; weldings, however, resulted in a 1.4. to 5-fold increase in PCDFs (il). Samples of used askarel taken frem transformers manufactured In the United States and Canada have also been analyzed for PC0F concentrations. For Instance, both bulk samples of used askarel and air In a transformer maintenance workplace were ana lyzed; results showed trace TCOF concentrations (0.013 to 0.116 ppm) In the fluids, and no detectable concentrations in the air (3). Two samples of used askarel were also analyzed by the National Institute of Environmental Health Sciences (21 Fluid A, obtained from a transformer of early manufacture but unknown extent of use, contained 0.7 ppm PCOF (66h Fluid 0, a bulk simple of used askarel obtained from a HONS 215230 3-14 transformer maintenance ooeration (66), contained 4,7 pom PCOF (74), The PCOFs found in Fluid B consisted primarily of octa-COF (4,5 ppm), and no detectable tetraTCOF. In 1979, Chittim and coworkers (_7S_) investigated the formation of TCDFs in transfor mer askarels, using gas chromatography with a oacked coluim. They reported TCDF concentrations ranging from approximately 0.004 to 4.7 ppm fn used askarels (Table 3-5). The amount of PCDOs, if detected, was usually less than 5 percent of the TC3F concentrations (_7_5). Small amounts of terphenyl, quaterphenyl, and chlorinated terphenyl were also detected In some of the askarel samples, but they were not quanti fied. The concentrations of TCOF in transformer askarels were found to increase with time in service. Furthermore, the apparent changes In TCOF concentrations caused by discharging or arcing were found to be negligible, most likely because of the short duration of the sparks or events causing failure, (t should be noted that the authors themselves acknowledged the limitations of the applied analytical method (l.e., packed chromatoyaphic coluim). tn 1962, the General Electric Company (GE) submitted a sample of used askarel (obtained from the Canadian General Electric Company} to both a university-affili ated and an industrial laboratory for PCOF analysis. Using high-resolution gas chromatography/mass spectroscopy (GC/HS), the industrial laboratory found PCOFs on the order of 1.0 ppm or lower (66). The university-aff111 ated laboratory found no PCOFs at detection limits of 10.0 ppm for tetri- and pentachloro isomers, and 1.0 ppm for hexa-, heota-, and octachloro Isomars (66). Subsequently, GE submitted four additional samples of used askarel to the same in dustrial laboratory for PCOF analysis (66). These samples had been obtained from in-service distribution transformers at a GE transformer manufacturing facility in the United States during the course of normal maintenance activities. The results of these analyses, presented In Table 3-6, show PCOF concentration ranges from 0.62 to 1.66 ppm In 40-year-old askarels; these PCOF concentrations are coxparable to the PCOF concentrations reported for unused Aroclors. The formation of PCOFs when askarels are exposed to sufficient oxygen and high temp eratures was dennstrated by conducting laboratory-scale pyrolysis at 500*C of two PCS mixtures (Aroclor 1254 and Aroclor 1016) and three askarels (Chlorextol, Pyanol, and tnerteen) (75). The results of these tests (Table 3-7) indicated the formation of high concentrations of PCOFs. Aroclor 1016 generated sfqnlflcantly 3-15 HONS 215231 Table 3-5 PCBi* ANO TCDF CONCENTRATIONS (ppn) IN USED ASKARELS (75) S IS S t i3 4 Grata 1 l-l 1-2 l-l 1-4 l-l 1-4 1-7 1-4 l-l 1*10 Ml LIST IH1 1*41 (HI 1*41 IMI 1*70 1*70 1*71 1*74 tan *r tr tr tr tr XT XT tP tP >10 tr 21.4 21.4 27 1 21.1 21 4 21.0 22.1 21 2 21.2 21.4 20.1 1 1 1 3 14 4 2 4 91 0/4*.0 1240 0.700 1 f tr 19.2 3.4 91 S/M 4 12*0 l.ilO 1.2 tr 14 0 2.1 90.1/4*.4 12*0 1 020 1 4 tr 14 4 2.* *2-4/47.1 12*0 0 7*0 1.0 tr 2.2 tr 31.1/11.4 1200 0 150 7 1 tr ' 0 tf l.l *0 tr 90 17.2/12,1 1240 0 2*0 21.1/70,4 1254 3 0 i.l tr ir 90 24-0/72 0 1254 0 470 7 4 tr tr 90 21 0/71 3 1254 3 2*0 i 4 10 10 90 24.4/71 | 1214 3 051 9 4 tp tr 90 10.1/1* 4 1254 3 041 trot* 2 2-1 2-2 2-2 1*47 m 1147 tp 1*47 tp 21.4 21.2 21.0 2-4 1*47 tp 21.4 2-1 IMS tp 27.1 2*4 1*41 tp 27.0 2-7 IMI tp 27.1 2-4 U4I tr 24.4 4 1 tr 14 1 3.4 49-2/94 | 1200 0.010 7 2 tr 14 1 1.2 44 1/91 7 1200 0 004 7 0 tr 14.4 3.2 50-4/41.4 12*0 0 007 1.4 tr 11 4 2.1 4*.0/41.0 12*0 0.010 7. a tr tr tr 19.4/44.1 1200 0 171 9.a tr 11.1 2.1 4*.0/*1.0 12*0 0.434 t.t tr 1.2 tf 1 0 tr tr tr 1339.4/44 f 14.4/11.4 0.110 ? 0.141 fire* 1 3-1 1-2 1-1 1-4 l-J 1-4 LM7 40 1*47 tr IM7 tp 1*47 tp IM7 tr 1*47 tr 21.4 21.0 2*. 2 27 4 21.0 21.0 9.a tf 7 0 tf 9.0 tr 7 I tr 7.1 tr 7 4 tr 14*1 3*4 49.2/14.1 14.4 1.2 90.1/41.4 tr tr 34.2/fl.l tf tr 11.2/44.1 tr tr 32.1/47 2 tr tr 12.1/17.2 1240 1210 1210 12*0 1290 12*0 0 411 0.141 0 444 3.421 31 3.1 fr-aie 4 4-1 1*47 40 2.| ' 2 tr 14.2 3.0 41.2/90.1 12*0 4 7 4-2 :*7 w 29 2 9 1 tr 14.0 2 1 M. 4/91.1 12*0 2 2 4-1 IMI 40 27.0 9.2 tr 14 4 3.1 90.2/4*.* 1210 3.4 i-4 1*71 40 27.4 9.2 tr 14.1 4.0 91.4/M.1 12*0 0 4 4-4 <* tr 11.4 11.4 tr 10.0 90 91.1/41.2 1254 3 3 4-4 tan 40 20.2 10.0 tr 0 90 30.2/ft.l 1254 3 0 4.7 Hat tr 20.1 *. tr tr a 10.1/41.4 12*4 3.0 * fCI* Hl)ciilriMM mn<n feane t: (l-l tfrae** t-tt) - rarlMla ttv-tn-iarrlct (ft). 2; (l-l tlra^M 2-) . cat tronfmn: vartMla tM'Mhv 'ittg (ft* MfaMli Cl (1*1 tlrawM 2-1) - ft trml*t*rt; rarlMla f**t(nlw riot (ftt Aaeanen C) Oraae 1: tart*1 flat* tym - J-t aM J-2 fwanal (Ctt) J-l an* 1-4 Chlaraital (ft) 1-1 m* >-1 Intrttt* (N). *: 4-2 "* *-2 - *<*ry **e trmlimn (tt): l*r*a variation in aeoer/votw ratio 4*) in 4-4 . trMfran I* let *a* ft 11*4 I* ftrv let (40. tt). 4-1. 4-4. 0*4 4*7 - nan fgran* I (CSC). Intrtaa* (4). t* CMoroilel (ft) tm(Kt*rlr) (ftu tjnt) Chlaracaapa (71). Cat - Cw4ln fitter11 dictric ft - farraatI-factor* H - ttltlWIMH 40 On eeotr! U4 fatter flactrlc (tyrant)) (Chlaraital) Inerte*a) tjraclor Inerta**) 3-16 MOMS 215232 Tib it 3-6 CONCENTRATION OF PCOFs IN ASKAREL TRANSFORMER FLUIDS FROM IN-SERVICE DISTRIBUTION TRANSFORMERS AT GE transformer manlfacturing FACILITT (7S) Sanple Nunmr Type of Transfor^r 3 Single phase - 100 kVA 2300 vt -- 115/230 115 gallons (435 liters) fluid 23 3 phase - 1250 kVA 1380 vt -- 480Y/277 490 gallons (1855 liters) fluid 44 2 phase - 375 kVA 2300 vt -- 550 31.5 gal ions (119 liters) 45 Slnple phase - 100 kVA 2300 vt -- 120/240 42 gallons (159 liters) `Pure Aroclor 1254- (control) Oate I ns tel led 1940 1947 1938 1939 Total PCOF (ppm) 1.1 0.62-0.82 1.84-1.86 0.64-0.86 0.62-0.82 < PCB 56.4 66.2 60.1 66-8 HONS 215233 3-17 Table 3-7 PYROLYSIS OF PCBs ANO ASKARElS AT 500*C (7S) PCB or Astarel Aroclor 1254 Hass Pyrolyted (">9) 25.0 40.0 Aroclor 1016 25.0 40.0 Pyranol 25.0 0.25 Inerteen 0.25 0.25 Cblorextol 0.25 0.25 Posltion of # Samole 0 C 0 c 0c c0 0 c 0 c 0c Q c 0c Q c Hess of Tetrachl prod!beniofurarts Formed Actual Hass (ug) 2.20 1.33 4.90 3.03 Per Gran of Starting Haten af (ug/g) B8.0 53.2 122.5 75.8 Total (<K) Aerage oi " Both Runs (Ufl/Q) 141.2 197.3 169.3 0.038 0.027 0.072 0.026 1.52 1.08 1.80 0.65 2.60 2.45 2.53 0.53 0.53 0.048 0.074 21.2 21.2 191.7 297.0 42.4 488.8 NA 0.030 0.030 0.033 0.003 118.3 121.9 133.7 12.2 240.2 145.9 193.1 0.023 0.025 0.021 0.041 93.5 100.2 82.8 165.7 193.7 248.5 221.1 * Sampling points: Q fimdlately after the reaction tube; C a cold trap further along In the reaction apparatus. 3-18 HONS 15234 lets PCOF than Aroclor 1254 when pyrolyzed. Thete results were to be expected since Aroclor 1016 has lower chlorine content, and thus does not contain as much of the higher chlorinated biphenyls (i.e., penta- or hexachtoro), which are the necessary precursors of TCDFs. As discussed above, PCB conversion to PCOF or chiorobeniene conversion to PCOO and PCOF might be possible in a malfunctioning transfomer or capacitor, provided that there is sufficient time at optimum temperature and/or oxygen conditions for their formation. Ordinary working conditions do not appear conducive to PC00/PC0F forma tion (66). As described m Section 2, electrical equipment PCB fires may begin internally or externally, If they begin internally In an airless transformer or capacitor (with either no head space or an inert atmosphere), the early stages are caused by elec trical energy in interval arcing. Under these conditions, the major combustion products are PCBPs; smaller amounts of PCDFs, PCPYs, and PCCYs are also found (C. Raope, Personal Communication). PCBPs were identified for the first time in the aftermath of a 1981 capacitor explosion in Stockholm, Sweden (2). If the equipomnt explodes or cracks and the PCBs are exposed to air, hi ?wr concen trations of PCOFs will form. If the fire begins externally, PCDFs are the major combustion product accompanied by PCPYs and PCCYs; no PCBPs will form. PCDOs will form only If chlorobenzenes/chlorophenols are present in the PCB or askarel. De tails of detected combustion products after different electrical equipment incidents are given in Section 2- Flnally, it should also be noted that most of the research on PCB/askare) conversion to PCOO, PCOF, and other chlorinated species has been performed using pure PCBs or askarel formulttions containing at least 50 percent PCBs. Very little research has been conducted on dilute PCB systems, such as contaminated mineral oils. Conse quently, there is little data available on the effects of dilution on PCB chemistry. HONS 215235 3-19 Section 4 CHEMICAL ANO biological degradation Very little research has been done on degradation mechanisms of dioxins and dibenzofurans. This Is particularly true of the chlorinated dlbeniofurans; there is almost no information published on degradation. A nunfeer of researchers have speculated on degradation pathways, but little actual research has been conducted. Consequently, the discussion mist center primarily on the chlorinated dioxins. CHEMICAL DEGRADATION Ozonation Ozone bubbled through a suspension of 2,3,7,8-TCOO In water and carbon tetrachloride for 50 hours achieved 97 percent degradation (_33). The dioxin was apparently sus pended as an aerosol combined with the CC14, which facilitated the ozone attack. Although not tested with dioxins, the combination of ozone and ultraviolet Irradia tion has been successfully used to degrade oentachlorophenol and DOT (33). The UV activates the molecules to a highly energetic state, making them more susceptible to ozone attack. Chloroiodide Destruction Recently, Botre, ft al. (77), treated contaminated soil from Save so, Italy, with several chloroiodides (a class of compounds derived from quaternary ammonium salt surfactant). In laboratory tests on residues frps the vacuus evaporation of 2,3, 7,8-TCOO in benzene, alkyl dimethyl benzyl ammonias (benzalkonium) chloroiodide achieved 71 percent 2,3,7,8-TCOD degradation. The use of l-hexadecylpyridinium (cetylpyridlnium) achieved 92 percent degradation. In tests on actual soil samples, benzalkonium chloride achieved only about 14 percent degradation after 24 hours. Mten benzalkonium chloroiodide wes added, an additional 38 percent was degraded. The chloroiodides apparently cleave the ether linkage. Alkaline Oehvdrochlorlnatlon Alkaline dehydrochiorination is a generic tens for several new chemical processes that have been developed to treat and dispose of hazardous chlorinated hy*-ocarbon 4-1 HONS 15236 wastes. However, only the process developed by the vertac Chemical Company of Hemphis, Tennessee, has been tested on TCOO and other chlorinated dioxins (78). The vertac process involves the dehalogenation of chlorinated dioxins with anhydrous alkali metal salts of polyhydroxy alcohols at atmospheric pressure, or by reacting PCODs, an alcohol, and a water solution of an alkali metal hydroxide. Vertac claims that 2,3,7,8-TCDD and chlorinated dioxins are reduced to essentially aero (78). Experiments have been conducted by vertac using 2,4,5-TCPh still bottoms, containing up to 250 ppm TCOD. A stoichlonetric excess (based on the halogen content) of alkali metal alcoholates of C4.g-alcoho]s, C^.g-alkanepolyols, and monoalkyl ethers was used. The reaction took place at a temperature of 140'C to 220*C for a time sufficient to convert the organic halogen into inorganic halides. The reaction product was essentially free of 2,3,7,8-TCDD (ppt range) and other chlorinated diox ins (i.e., more than 99.95 percent destruction) (79). Catalytic Reduction with Iron Chlorides In 1972, experiments were conducted to investigate the potential for separation of esters of 2,4*0 from 2,4,5-T in Agent Orange by fractionation (78). Overheads <re found to contain 93 percent 2,4-0 ester, whereas bottoms comprised 93 percent 2,4,5T ester. It was not possible to determine TCOD closure by materials balance, as the stainless packing and steel colum initiated some catalytic destruction of the TCOO. The bottoms fraction (2,4,5-T) contained more than 93 percent of the total TCOO, while the overhead fraction (2,4-0) contained 0.15 ppm TC00. These experiments dem onstrated that an effective and efficient separation of the esters was feasible (28). Catalytic Oxidation wi th Ruthenium Tetroxide Ruthenium tetroxide has been successfully employed in the laboratory to destroy 2,70CDD and TCOD (77). Reaction kinetics were studied using solutions containing 70 to 72 ppm PCDO in carbon tetrachloride and 2 to 4 ppm ruthenium tetroxide as a function of temperature. The results indicated a 2.4-fold increase m the r4te constant per 10*C increase in temperature. At 7D*C, the half-life of TCOD was found to be less than 15 minutes, and that of 2,7-OCOO, approximately 10 minutes. MOMS 215237 4-2 Micellar Catalysis with Chloroiodldes The most premising results have been obtained with benzalkomum dlchloroiodide and cetylpyridlnluw dichloroiodlde (77). The low solubilities of these compounds *re increased by the use of micellar solutions of the same surfactants, TCDD degrada tion products included chloroohenols, phenols, and 2-phenoxy-chlorophenol. These findings Indicated that micellar catalysis (which Is carried out In the absence of harsh acfdlc conditions, at ambient temperature, and without the use of light energy) results In a cleavage of the ether link. Chlorolysls The chtorolysis process operates with chlorine gas as the oxidant at a temperature of 600* C and a pressure of 2500 osig (170 atm). The Noechst-Uhde Corporation, under contract to EPA, conducted an experimental study to assess the feasibility of the chlorolysls process to convert typical chlorinated solvent wastes and Agent Orange to useful products such as carbonyl chloride, hydrogen chloride, and carbon tetra chloride (78). The primary Interest of the study was the destruction of the TCDD. The pretreated military defoliant Initially contained 14 to 18 ppe TCDD, whereas the carbon tetrachlorIde product contained no aeasurablc TCDD at a detection limit of 1 ppb (80). PHOTOLYSIS There are basically tlree conditions necessary for the photoreduction of chlorinated aromatic compounds (81,, 82): e Presence of a hydrogen donor, usually in the solvent, e ultraviolet (UV) light of the appropriate wavelength, e Absorption of the light. In sunlight, only these expounds absorbing UV light above 290 nm can be expected to be subject to photode^adatIon. This Includes the PCDOs and PCOFs. TCDD photolysis proceeds via stepwise photoreductlve dechlorination as follows (_*6, 81.): MONS 215238 4-3 A hydrogen donor is essential ; as a result, degradation is fast only in organic media, slow in water, and practically nonexistent on dry, inorganic surfaces (*6). Most of the photolysis research has been conducted on organic solvents, such as methanol, hexane, diesel oil, or phenoxjether (46, JI_, ,83). In these solvents, the lower chlorinated isomers are extremely unstable to light. For instance, Crosby, et al. (46), have shown that 2,7-dlchloro-, 2,3,7-trlchloro-, and 2,f,7,8-tetractilorodlbenzodloxin in mathanol will totally degrade within a few hours. The octachloro Isomer was nuch more stable. Shortly after the Seveso incident, it was demonstrated that olive oil Is a suitable hydrogen donor (64). Total reductions in TCOD levels In excess of 60 percent were observed 48 hours after treatment with 400 1 of olive oil emulsion per ha of contam inated soil. Furthermore, less than 0.07 percent of the applied olive oil was found In the leachate. Indicating that TCOD could not be transported to lower geologic strata, even If It were totally dissolved In the oil film on vegetation or the ground. 8otre, et al. (85). evaluated various solvents to solubilize pure 2,3,7,8-TCDD and sables of contaminated soil from Seveso in aqueous solutions containing cationic, anionic, and nonionic surfactants prior to photolysis. The cationic surfactant, 1hexadecylpyrldlnlimi chloride (HOPC), was found to be effective in solubilizing TCOD and In enhancing photolysis. The total destruction of 8 ug/ml 2,3,7,8-TCDD In 0.091 HOPC was accomplished In 4 hours. Stanford Research Institute reported a half-life for gas-phase TCOD in sunlight of 5 to 24 days (55). Mhen samples of TCOD wire placed on dry soil and glass plate and Irradiated with UV, no degradation was observed after g6 hours and 14 days, respec tively (83). TCOD In the crystalline state resists photolysis (81). Thus, the presence of the hydrogen donor Is essential to rapid photolysis. In general, the rate of dioxin photolysis Increases as the number of substituent chlorine atoms decreases (83). According to Pllmapr, at al. (S3), even the dioxin nucleus will photodegrade In aothanol. On the other hand, Crosby (,81.) reports that monochi or odlbenzodl oxl n is essentially stable to photolysis. In 1981, 4300 gallons of wastes containing approxlaetely 343 ppm each of TCOD (7.5 *9)i trlchlorophenols (50 to 55 percent), and ethylana glycol dtrlvatlves (45 to 50 percent) were subjected to 4 coeaprdal-scale batch trtatepnt UV-dtgridatlon process HOMS 215239 4.4 for TCDD destruction. TVs destruction process involved the two following major steps: (1) aultlphase extractlon/neutralIzatlon with sulfuric acid/n-hexane/caustic soda (up to eight multlole extractions of the hexane, acid, and oily phases); and (?) photo lytic reduction using UV-irradl atton and Isopropyl alcohol as a proton donor. The process was found to be effective in reducing the mlttil TCDO from 7.5 kg to less than 100 g (destruction efficiency of 98.24 percent). The picture with regard to dlbenzofurans is less dear. Crosby, et al. (46). re ported that PCOFs follow the same degradation pattern as PCOOs, with higher chlori nated Isomers being more stable. Hutzinger (86) reported that both 2,8-dtch) oroand octachlorodlbenzofuran In methanol and hexane photodechl or mated rapidly. fluser (45) stated that higher chlorinated PCOF isomers photodegrade faster. INCINERATION There has been little research conducted on thermal destruction of PCDD, and even less on PCOF. Different researchers have reported 2,3,7,8-TCDO destruction at SOO'C (40, 55). In general, however. Incinerator operating conditions currently consid ered adequate for TCDO destruction are a temperature of at least 1000*C and a dwell time of 2 seconds (33). At 700`C and a dwell time of 21 seconds, only 50 percent destruction was achieved;, at 800*C and a dwell-time of 21 seconds, 99.5 percent de struction was achieved. In 1972, tests were conducted to evaluate the feasibility of Agent Orange Incinera tion (78). The Incineration test unit consisted of a SUE* burner/incinerator and reaction tailpipe, a Venturi scrubber, a scrubber collection bank, and a pas sam pling system. Both exhaust gas and scrubber liquid effluents were samoled and ana lyzed for TCOO. The test results showed that TCOO was not present cither in the combustion gases or In the scrubber liquid at the detection limit of 15 ppb (78). Recently, a truck-mounted nob lie waste processor, develooed by Pyro-Magnetlcs Corpo ration, was evaluated at the diversity of Tennessee Space Institute in Tullahoma, Tennessee (87). The mobile incinerator was fed a 41.7 percent PC8 waste at the rate of 222 1/hour In four tests totalling 14 hours in duration. Average destruction and coxttustlon efficiencies of PC8s were 99.9999 and 99.9979 percent, respectively. The Incinerator outlet temperature was reported to average 1245'C, and the residence time was 2.9 seconds. The 2,3,7,8-TCOO was detected In only one of ttree test burns, and averaged 14 percent of the total TCOO; the 2,3,7,8-TCDF Isomer averaged 27 percent of the total TCDF. HOMS 215240 4-5 Ackerman end coworkers (_30) have developed guidelines for land-based incineration of PC8s and related materials; the criteria are summarized in Table 4-1. For non. liouid PCBs or materials contaminated with PCDFs and/or PCOOs, there is an addi tional requirement: the mass air emissions must contain 0.001 g PCS/kg PCS fed to the incinerator Table 4-1 CW8USTI0N CRITERIA FOR PC8s ANO PCDF/PCOO- CONTAMINATEO WASTES 1. Maintenance of introduced llouids for 2-sec duel 1 time at 1200*C (+10D*C) and 3 percent excess oxygen in the stack gas, or for l.5-sec dwell time at 1600*C (100*0 and 2 percent excess oxygen in the stack gas. 2. Combustion efficiency >99.9 percent based on lOOCCO^VICCOj] + [CO]). 3. Rate and quantity of PC8s fed to combustion chanber oust be measured and re corded at regular Intervals (<15 min). 4. Incineration temperatures continuously measured and recorded. 5. Monitoring of stack emissions products for: a. b. c. d. e. f. PC 8s g. PCDFs and PCODs h. Total particulate matter. 6. Automatic shutoff of waste flow when: a. Combustion temperature decreases to below conditions stated in Item 1 above. b. Failure of monitoring operations stated In Item 5 above. 7. Caustic scrubbers used for HC1 control (or an approved alternate method). To date, M/T Vulcanus is the only incineration vessel that has been employed to in cinerate TCOD-contamlnated materials (78). The M/T Vulcanus Is a double-hull, dou ble-bottom tanker specifically designed to Incinerate chlorinated hydrocarbon waste HONS 215241 4.6 products. Waste to be processed must be liquid in order to be pumoed. althougn solid substances up to 2 in (5 cm) in site may be included. T*o incinerators are installed aboard, eacti having an overall height of 34.25 ft (10.45 m) with an inner dimension of 15.75 ft (4.B m) for the combustion chancer. The waste feed rate is about 12.25 tons/hour (12.5 tonnes/hour), and the residence time is approximately 1.0 second at 1500"C (7B). biological degradation Investigations have shown that dioxins exhibit a relatively strong resistance to biodegradation (36. 50). Most of the work on dioxins has been focused on 2.3,7,3TCOD because of its extreme toxicity. Approximately 100 strains of microbes, wnich had previously shown the ability to degrade persistent pesticides, were tested. Only five strains were shown to have the ability to degrade 2,3,7,8-TCOO (36, IB8). It is possible that the lesser chlorinated PCOOs are more susceptible to biodegrada tion, as Is the case with PC8s (50). TCOO was found to be practically immobile in soil, and adsorbed strongly on lake sediments (89 , 90, _91^ ^92. ^3. J4). Although 2,3,7,8-TCOO may be genuinely metabolized m aquatic ecosystems, the rate is ex tremely slow (62). Klecka and Gibson (1979) have reported that unsubstituted dlbento-P,-d1 oxm can be readily nmtabollzed by a mutant strain of Pseudomonas (sp. N.C.I.8. 9816 strain II) when an alternative source of carbon, such as salicylate. Is available (_36). The dioxin molecule was metabolized first to a ci_s-l,2-dihydroxy-1,2-dlhydrodibenzo[l,4]d1oxin (rxn 11) which was subsequently dehydrated to yield 2-hydroxyd1benzo[l,4]-d1oxin (rxn 12) as the major metabolite. They also reported finding no organisms capable of utiliz ing dlbento-p-dloxln as the sole source of carbon. MONS 215242 4-7 Cano"' and coworkers (^95_) evaluated microbial degradation of TCOO 'n soil samples from "Zone A' at Seveso. Soil samples containing an average of 0.1 ppm TCDO were analyzed in the presence and absence of organic compost. The results indicated a 25 percent reduction in TCDO concentration in 480 days. Studies by Kearney, et al. (96), have indicated that the average TC00 remaining in the soil after 1 year of weathering was approximately 50 percent at all concentrations tested (1 to 10 ppm). However, it has been determined that microbial degradation is not a major factor m the environmental degradability of these toxins. The predominant mode of elimina tion of this conpound within the environment appears to be In the photodecomoosition by sunlight (88). On the other hand, research by Hutter (7) and Young (98) indicates that microbial degradation of TCDO is very slow under the most optimum conditions, leading to the conclusion that dioxins are essentially nonblodegradable. furthermore, these re searchers suggest that the half-Hves for dioxins In soil observed In earlier exper iments are actually the result of drastically decreased extractablllty, and do not reflect a loss of material from the soil. Recent research has shown that dioxins are metabolizable by certain mammals (99). The biotransformation of TC00 In dogs appears to be a detoxification process. Experiments with guinea pigs showed that the acute toxicity of the metabolites was at least 100 times lower than that of the parent compound- Conversely, when predloxins were administered to animals, there was no Indication of netabolIsm-induced dioxin formation (C. Rappe, Personal Communication). Little research has been conducted on dlbenzofuran biodegradation. However, the metabolism of chlorinated biphenyls may produce penta- and hexa-COFs (64). The re searchers have suggested that the PCDFs were synthesized via ortho-hydroxylatlon in association with chlorine miration, and have further Indicated that It may have a substitution pattern at positions 1, 2, 3, 7, 8, and 9. This newly discovered phe nomenon is of p-eat toxicological significance. There is also evidence to suggest that tetra-CDFs may be formed by liver metabolism In rats. These COFs are potenti ally wch nore toxic than the parent chloroblphenyls, even if their production is via a minor metabolic pathway. The metabolism of PCSs has often been regarded as a detoxification process. This Is particularly so tdien the metabolites arc conjugated and excreted. HOMS 215243 4-8 Section 5 ENVIRONMENTAL persistence with their high tonicities, the environmental fates of PCODs and PCDFs are of in* creasing importance. Unfortunately, little is known of the environmental benavior of most of these compounds. Only 2,3,7,8-TCDO has been studied in any detail, and then often with inconclusive results (100). To what degree the findings with TCOO are applicable to other PCODs and PCOFs is largely undetermined. Primarily as a result of the Seveso incident, most of the environmental research nas centered on the persistence of TC00 in soil. In general, TC00 is highly persistent In soils (32). According to 01 Domenico, et al. (101). there is a high initial loss rate due to volatilization and photodecomposttlon of TC00 present on the surface, followed by a very slow steady-state loss rate. They estimate that after 10 jeers, 30 to 44 percent of the TCOO present after the first half year would still be in the soil. Since other researchers have Indicated that up to 50 percent can be lost in the first year, this emphasizes the eitremely slow loss rate m subsequent jears (55. 96). Once the TCOO has been washed Into the soil, it is protected from the influence of solar photolysis and volatilization (102, 103) It is not easily washed out of soils, being virtually immobile m all but very sandy soils (_7). The role of such processes as erosion as a transport mechanism is unknown (0). TCOO is not taken uo by plants to any significant dearee U, 5), and there is relatively little biomagmflcatlon In aumeals (50). As noted in the previous section, these compounds are not readily biodegradable. Stehl, et al. (104), have suggested that 2,3,7,8-TCOO is probably stable toward air oildatlon. There appears to be only one significant environmental decomposition mechanism: photolysis. As noted earlier, however, a hydrogen donor Is necessary for this reac tion. Consequently, TCOO deposited on dry surfaces may be resistant even to photol ysis. Crosby (81) denonstrated at Seveso that spraying a contaminated environment with an Innocuous organic solvent (In this case, olive oil) can greatly enhance TCOO decoevosltlon. MOMS 215244 5-1 If TCOO I* washed into a water body, it will strongly adsorb onto bottom sediments (55). There is some evidence to suqgest that PCOOs and PCDFs will bind stronoly to any particle, ho data is available on the solubilities of the sorbed PCOOs and PCOFs. Neither PCOOs nor PCDFs are particularly soluble in water, and solubility decreases with increastno chlorine saturation (). How the solubility of PCOOs is affected by the presence of colloids or solvating agents In natural waters is un known ('50). However, there is some evidence that TC00 bioaccumulates, suggesting at least a limited mobility throuoh the food chain (^, 1QQ). Fish do not seem to biomagntfy TCOO to any great extent through food consumption (0). However, there is evidence to Indicate that bioaccumulation from the water vector can exceed 6000 times the water content (105) Nevertheless, different Isomers be nave differently, and the extent and impact of these differences require further investigation (50). Table 5-1 provides evidence of the lower solubility of the higher chlorinated iso mers. In addition. Table 5-1 shows the benavior of different isomers in fish. Several researchers have speculated on various environmental prooertles of TCOO, based on the properties and fate of other similar organic conpounds (see Table 5-2). These have then been used to predict persistence and fate in a model aquatic ecosys tem. Where testable, these predictions have been reasonably accurate. However, m the absence of more extensive test data, these constants remain speculative. HONS 215245 5-2 Table 5-1 PCOO AMO PCOF CONCENTRATION RANGE IN HUDSON ANO HOUSATONIC RIVES SEDIMENTS AND FISH FROM 4JOOS ONO (106) 4 ! CS rh KVi Z'i = '7 CIJ 1 ava- 10* . 44 io 10 I.JJ9 J,)H 3 19,990 10 700 10 If] 10 - 377 10 J.`3 10 1.010 04l <0 00) 0 301 0 19 0 11 3.37 MOQtff >9"J wH0to Vir Mqvq t i :* 0 94 0 4* <0 009 <0 009 * do o# otttetM. Table 5-2 SIMMARY OF PREDICTED PARTITION, TRANSFER. ANO DEGRADATION CONSTANTS FOR 2,3,7,3-TCDD (SO) witttimtiM cs*c) MWUlfiH,[*!**, Jim) tttuMtiwt Olya 3<m illMIM irtittpi (IQS 4ry*ie) (IS arpale) rpimftr If Ndr (i (lOOl arpaic) (101 4ry4DWj ,. <uccuailt4ii Partition Trmftr (1*1 *-*J UOMM ciaarann FrptlOO OlfH 5-3 1.7 ( 102 0 5 1.1 i 10* 1.1 i 10s 1.0 < 10? 1.0 I 10* ].) 10* l. 102 9.9 t 10`* 0 HONS 215246 Section 6 PCOO/PCDF/PCBP TOXICOLOGY INTRODUCTION The following section discusses the toxicological effects of PCOOs, PCDFs, and PCBPs on laboratory animals and man. On a molecular basis, 2,3,7,8-TCOO Is perhaps the most toxic synthetic molecule knowi to man (_33). Only bacterial exotoxins (i.e., Botuimum Toxin A, tetanus toxin, and diphtheria toxin) are more potent ooisons (107) . Both higher and lower chlorination reduce toxicity, but the toxic symptoms of the various isomers are similar, the only difference being the amount reguired to oroduce a given effect. The isomers with the highest acute toxicity are 2,3,7,8-tetra-CDO, 1,2,3,7,8-pentaCOD, 1.2,3,4,7,8-, 1,2,3,6,7,8*. and 1,2,3,7.8,9-hexa-COD. 2.3.7.8-tetra-CCF. 1,2,3,7,8- and 2,3,4,7,8-penta-CCf, and 1,2,3,4,7,8- and 2,3,4,6,7,8-hexa-COF (see Figure 6-1). This figure clearly illustrates the current thought that full lateral substitution with Cl In the 2,3,7,expositions leads to high toxic potency. Most of the research has centered on the 2,3,7,8- Isoamrs. ANIMAL TOXICITY Rodents Comparative oral lethal dose values for various PCOOs In mice and guinea pigs clearly shorn that 2,3,7,8-TCOD and 1,2,3,7,8-penta-COO isomers are the most toxic (108). The toxic effects Induced by PCOOs vary both quantitatively and qualita tively among different species (33_. 108. 109. 110). The major characteristics of lethal doses of 2,3.7,8-TCDD In all animal species are progressive weight loss and general weakening, with death occurring within a few weeks or months (111). Degen eration of the liver is also a comm pathologic effect (112). hepatic necrosis produced by 2,3,7,8-TCOD is probably a contributing cause of death in rats and rab bits. However, the number of deaths attributable to liver insufficiency is loimr in mice, and minimal in guinea pigs (108). 6-1 HONS 215247 Flguro 6*1. Tht Most Toxic PCDO ud PCDF Isamrs HONS 215248 There is minimal damage to the liver of guinea Digs, the species most sensitive to 2,3,7,8-TCOO toxicity. This suggests th*t 2,3,7,8-TCOD is probebly effecting some fundamental process in animal celts, which leads to a oenerat dysfunction of those celts (1111. A targe percentage of 2,3,7,8-TCOD persists in tne liver In unmetaboHzed form, par ticularly concentrated m the microsomes and microsomal fractions (fragments of ribosomes and endoplasmic reticulum) (.33,, 109, I la). These findings suggest that the unmetabolfzed 2,3,7,8-TCDD, rather than a metabolite, is responsible for the observed toxic effects (1151- Various experiments conducted to date have failed to provide the exact biochemical mechanism(s) of 2,3,7,8-TCDD lethality. Adverse effects on the following processes do not appear to be the mechanisms by which 2,3,7,8-TCOO exerts its toxicity (33. Ul): e ONA synthesis, e Protein synthesis, e ATP production. e Oxidation-reductionstate of cells. e Action of thyroid and glucocorticoid hormones. e General absorption of carbohydrates, amino acids, and fatty acids from the Intestine and the transfer through cell membranes. e Aerobic and anaerobic metabollsat of carbohydrates, amino acids, and fatty acids, e fatty acid composition of tissues. At this time, the most reasonable hypothesis Is that 2,3,7,8-TCOO binds to a recep tor in the cytosol (soluble portion of the cytoplasm) fraction of cells. Is trans ferred Into the nucleus, and Increases or decreases the synthesis of a critical proteln(s) or enzyme(s). This Increase or decrease In crucial protein or enzyme levels eventually leads to a general dysfunction of a nueter of different cell types, re sulting In death (111), However, the receptor theory may not explain the chloracnegenlclty of TCOO, Receptor proteins have not been Identified on skin cells. The lower toxicity of 2,3,7,8-TCOO In rats, as compared to other marnmals, suggests some degree of metabolic transformation of the parent compound (33, 115. 116. 117). MONS 213249 6-3 The concentration of dioxin in the liver of ret* may be five times greeter then tnet found in their body fet, Concentretion* within the kidney, thymus, and soleen are 1/12 to 1/50 of thet found in the liver (^2). The substance which is not retained is excreted unchanged with the feces (60 percent) vie the bile fluid. A smell amount (5 percent) is excreted in the urine as a water-soluble conjugate (1091. This suggests that 2,3,7,8-TCDO metabolism may be occurring in the rat, since waterinsoluble compounds, like dioxins, are excreted predominantly, if not exclusively, through the feces (33, 211) Female albino rats (Porton strain), given a single oral dose of 200 ug/kg 2,3,7,8TCDD (their approximate L0-50), died within 3 to 9 weeks following a period of weight loss and depressed food intake (116). Within 24 hours after the dose was administered, there vere changes In the hepatic parenchymal cells, including a pro liferation of the smooth endoplasmic reticulum, and alterations in the metabolism of various substrates. Approximately 4 weeks after the 2,3,7,8-TCOO was administered, giant multi nucleated cells were found within the area of the centrilobular tone (116). These cells were probably formed by the fusion of preexisting degenerate parenchymal cells (109. 117). This hypothesis Is supported by the following evi dence: e ho mitoses occurred within these cells. O' These cells tend to follow the line nf trabeculae. a Isolated bile canallcull were occasionally observed to be surrounded by the cytoplasm from the multinucleated cells. e Adjacent mononucleated parenchymal cells showed areas of apparent fusion of their plasma meeDranes, In addition, there were Increased numbers of lysosomes, loss of microvilli fron the bile canallcull, and proliferation of the smooth endoplasmic reticulum in the tentrllobular parenchymal cells (116). These lesions disappeared in surviving animals over a period of time. Hlstochemlcal 1y, there was a centrilobular loss of ATPase activity within 3 days. This lots extended to the periportal area by the sixth week (116). Biochemical studies on plasma membranes have indicated that the ha*/K* ATPase is unaffected, whereas the K*/Hg+* ATPase activity Is drastically reduced (109, 116). HONS 213250 6-4 It is known that the K*/Mg*^ ATPase of the sarcoplasmic reticulum reauires a lipid for activation; the degree of activation can be affected by the type of lipid in volved (116) Toxic levels of PCDD result in a chronic increase in triglycerides and developnent of a fatty liver {1081. It is possible that the initial disturbance of the oxidative enzymes of the endoplasmic reticulum leads to alterations in endo genous lipid nctabolism, with a conseouent change in the structure and function of the parenchymal cell membrane. These changes could lead to hepatic insufficiency with toxic effects observed macroscopically. 2,3,7,8-TCOO and other halooenated PCOOs stimulate a number of enzyme activities, most notably within the liver (1Q8, 111, II*. U, 119). Hepatic 6 aminolevulinic acid (ALA)* synthetase and aryl hydrocarbon hydroxylase (AHH)** are induced by the presence of PCDOs, particularly 2,3,7,8-TCOO (108. 120). The PCDOs which have been found to induce ALA synthetase appear to have two common properties: (1) halo9en atoms occupy at least three of the four ring positions (l.e., 2, 3, 7, 8); and (2) at least one free nonhalogen ated carbon atom is unoccupied (108, 118). Available toxicological data indicate that those PCDOs that are not toxic generally do not in duce ALA synthetase; those which are lethal, even at extremely law concentrations, induce the enzyme. 2,3,7,8-TCOO has been found to be many orders of magnitude more potent than any other known inducer of ALA U3, 118). microsomal monooxygenase (e.g., AHH) is a menbrane-bound enzyme complex which cata lyzes the oxidative (and for certain substrates, reductive) metabolism of numerous xenoblotlcs and endogenous lipophilic compounds (118). 2,3,7,8-TCOO is a highly lipophilic macromolecule (,120). The Induction of AHH by various PCOOs is earned out by the same mechanism as that found for the induction of ALA (118). The hypo thesis that the Induction of ALA and AHH by 2,3,7,8-TCOO is mediated by the interac tion of the ligand with an induction receptor (the product of a regulatory gene that cnblnes with a small molecule to Initiate gene expression) Is supported by the fol lowing evidence (118)1 a The potency of 2,3,7,8-TCOO In evoking a dose-related Induction of ALA and AHH activity. e Chemical specificity. * The initial and rate-limiting enzyme in heme synthesis. * A cytochrome P^-450 mediated monooxygenase. HONS 215251 6-5  SIological specificity (i.e., 4 specific type of microsomal monooxy genase activity is induced m some, Out not all, tissues). If tne hepatic cytosol-binding species is in fact the induction receptor, a oossible mechanism by which the receptor ligand complex in the cytosol initiates an increase m nuclear activity would involve tne translocation of this complex into the nucleus in a manner analogous to the mode of action ascribed to steroid hormone receptors (33, 118). Theoretically, the most logical pathway of 2, 3,7,8-TC00 metabolism is by hydroxylatlon at the 1, 4, 6, or 9 position of the molecule by AHH, The hydroxylation of aromatic rings in positions adjacent to halogen atoms is an uncommon reaction of the cytochrome P-448 monooxygenase system. This most likely accounts for the metabolic stability of 2,3,7,8-TCOO within mammalian systems (115, 116). However, it has been postulated that 2,3,7,8-TCOO moves into a cell and binds to a specific cytosolic receptor. The receptor-dioxin complex then moves into the nucteus of a cell, where it Initiates the synthesis of a specific messenger RNA, which directs the synthesis of cytochrome P-448, Other 2,3,7,8-TCOO molecules can then react with newly formed cytochromes, possibly to produce reactive Intermediates that reduce or imoeir cellu lar metabolism, gradually Inducing cell death (^3, 120). These metabolites may be excreted as innocuous by-products which may affect specific cells in other organs, or may act as carcinogens or cocarcinogens (33). If the activity of AHH were increased, more 2,3,7,8-TCOO would be metabolized. If 2,3,7,8-TCOO, rather than a metabolite, is responsible for the toxicity, then in creased metabolism by the P-450 monooxygenase system should lead to a decrease m toxicity. Conversely, If a metabollte(s) Is responsible for the toxicity of 2,3,7,3TCOO, then an Increase In metabolism may result In an Increase in toxicity (115. 116). Several studies have shown that 2,3,7,8-TCOO induces many enzyme systems while suppressing others. 2,3,7,8-TCOO Induces the following enzymes in addition to those previously mentioned (33, 108, 110): e HOP glucuronyl transferase. Aldehyde dehydrogenase. e Glutathione transferase 8. DT-dlaehorase. a Btnzopyrene hydrolase. MONS 215252 6-6  Glutathione S-transferase. Ethoxycoumarin aee thy Use. 2,3,7,8-TCDD reduces the activity of the following eniynes (891: e UDP-glucuronic ecid pyrophosohatase. - D-glucuronolactone dehydrogenase. L-gluconate dehydrogenase. A sublethal dose of 2,3,7,8-TCOO in rats will cause a temporary increase in trigly ceride and free fatty acid levels, with a persistent decrease of sterol esters. Lethal concentrations result tn a fatty liver, increased serum cholesterol esters, and free fatty acids, with little change m triglyceride levels. These changes are due m part to the damage sustained by lysosomes (_33). The decrease m the acid lipase activity, derived from damaged lysosomes, probably accounts for the increased levels of cholesterol esters within the liver. Therefore, another mechanism by which 2,3,7,8-TCOO may exert its toxic effect is by the increase of lipofuscin pig ments (33). Lipid peroxidation, which precedes the formation of polymeric lipofuscins, is known to seriously damage meebranous subcellular organelles, including lysosomes (^33, 108, 121). In one or more species of laboratory animals, bone marrow hypoplasia, testicular de generation, renal pelvis and urinary bladder hyperplasia, and hemorrhage m the in testines and adrenals have been observed (108). Chronic administration of low lev els of 2,3,7,8-TCOO to rats increases the incidence of neoolasia (tumors) (108) The high incidence of tumors in rats fed subacute levels of 2,3,7,8-TCOO (i.e., I ppt to 5 ppb)* suggests the carcinogenic potential of the compound (112). Lung turners that had been observed were characterized by squamous metaplasia of the bron chial epitheliimi and keratlniution. Lymphocytic leukeerta wes associated with a massive enlargement of the spleen. The most malignant tumors were observed m the lungs, kidneys, liver, and the skeletal miscutature. Other neoplasms were found in the brain, skin, testes, and ears (33, 108. 112). The only abnormalities of the hematopoietic system noted in rats have been hemoconcentration and throxbocytopenia * 1 part per tril Hon 10"^ g TCOO/g food. 1 part per billion 10"9 g TCOO/g food. 6-7 HONS 219253 (increase tn the relative volume of red blood cell* per unit volume of circulating blood and decreased nuneer of platelets per unit volume of blood) (33). The terms "embryotoxici ty" and "fetotoxici ty" denote all transient or permanent toxic effects induced In an embryo or fetus, regardless of the nmcnanlsm Involved (33). A special fetotoxic effect is teratogenicity, which may be defined as an abnormality (likely irreversible) caused by impairment of an activity *1ch is typi cal of embryonic or fetal development (_33, 122). This effect may either be obvious by macroscopic appearance or `hidden' (micromorphological defect or mental abnormal ity). 2,3,7,8-TCOO Is a specific teratogen, interfering only with a few soecial de velopmental processes (122). Recent studies have shown that 2,3,7,8-TCOO can nave a dramatic effect upon the hor mones involved In reproduction (33). As little as 0.3 ug/kg of 2,3,7,8-TCDD in jected Into mice was found to Increase the frequency of cleft palate (a teratogenic effect) (33, 122). This is by far the smallest effective dose of any teratogen known (122). Oral administration of 2,4,5-T containing dioxin to golden namsters on the sixth to tenth day of gestation resulted In an Increased absence of eyelids (33). Other fetotoxic effects of 2,3,7,8-TCOO Include thymic atrophy, fatty infil tration of the liver, general edema, delayed head ossification, low birth weight, fetal reabsorption, and kidney abnormalities (33). fetal mortality, early and late fetal reabsorptions, and fetal intestinal hemormaglng occurred in rats given a 2,3,7,8-TCOO dosage as low as 0.125 ug/kg U3). The fetal kidney anaaaly can be best described as a renal papilla which is markedly reduced In size, resulting in an enlarged renal pelvis (.108). This may progress Into hydronephrosis during metanephric kidney formation (123). The teratogenic effect of 2,3,7,8-TCOO may be a result of mutagenesis by Intercalation of the parent compound Into DKA, or by inter calation and covalent binding of the mttabollte, analogous to findings of tests con ducted with acridine and amlnofluorene compounds (120). Information i^llcatlng 2,3,7,8-TCOO as a mutagen Is both scarce and conflicting (33). Derivatives of dioxin have shown negative results when tested for dominant lethal effects In rats, and only slightly positive results when tested for their caoaclty to produce chromosomal aberrations in bone marrow cells of rats (.33, 108). Studies on mice Indicate that sub lethal doses of 2,3,7,8-TCOO increase tne nunper of erythrocytes (and their cell volume) and neutrophils, and decrease hemoglobin, serum MOMS 215254 6-8 protein, end globulin levels. The number of blood platelets decreases, resulting in diminished dot reaction (109). Another effect of 2,3,7,8-TCDD is its interaction vith iron metabolism (_33). Sets given 1.7 ug of the compound mtragastncally have shown a twofold increase in the serosal transfer of iron, whereas no change was observed in mucosar iron uptake. However, it appears that this iron deficiency protects mice from many of the toxic effects exhibited by 2,3,7,8-TCDD. Mice rendered fron-def icient were found to be protected from elevated porphyrin levels and liver damage. Since mixed-function oxidative enzymes were elevated m iron-deficient mice, it was speculated that the depleted storage of iron within the tissues was responsible for the observed reduc tion in toxicity (33). 2,3,7,8-TCDO, 2,3,7,8-TCOF, and 2,3,6,7-TC8P are the approximate fsostereomer of 3,4,3',4'-TC8. It is thus believed that the structural configurations that render TCOOs, TCOFs, and TC8Ps toxic are analogous to those for TC8s. Namely, 3,4,3' ,4'TC8 may assume a planar or nearly planar configuration with its four chlorine atoms projecting froei the lateral ring positions, similar to 2,3,7,8-TCOO. However, if chlorine is substituted in a position adjacent to the biphenyl bridge (e.g., 2,4,3', 4'-TC8), this sterically blotters the biphenyl rings and prevents planarity. Such nonplanar PCS analogs are incapable of initiating AHH activity (124). Thus, among the halogenated biphenyls, there are two structural retirements for the induction of AHH: (1) the presence of at least two adjacent halogen atoms in the lateral position of each benzene ring (e.o., positions 3,4,3',4' or 3,4,5,3',4',5'), and (2) the absence of halogen atoms adjacent to the biphenyl bridge (positions 2, 6, 2', 6'). The effects of different biphenyls on AHH are shown in Appendix 8. Birds It has been shown that there is a very good correlation between the potency of PCDOs and PCQFs to induce AHH activity and their capacity to produce chloracne, thymic atrophy, and edema in chickens (124). Histopathological examinations of birds exposed to 2,3,7,8-TCOO have revealed atro phy of the spleen, involution of the thywus, decreased nuefcers of lymphocytes (par ticularly in the bursa of Fabricius), pulmonary edema (i.e., myocardium), and necro sis of the liver (). The disease known as "chick edema" has also been reported in TCOO-exposed birds. This disease is characterized by the accumulation of serojs fluids in the ascites, anascara, and pericardial sac (125. 126) One study sho*md HONS 215255 6-9 that 3-day-old chicks treated mth doses of 1 and 10 uo of 2,3,7,8-TCDD/kg body weight developed chick edema f 125); the chicks also exhibited dyspnea, accumulation of mucus in the mouth, and enlargement and mottling of the liver (126), Alterations in drug-netaboliimg enzymes such as the cytochrome P-450 system and the accumula tion of porphyrin were also noted. The biological potency (EOjq) of 2,3,7,8-TCDD and various biphenylene and biphenyl congeners was estimated from a log dose-response curve for the Induction of hepatic aryl hydrocarbon hydroxylase activity in a chicken embryo (124) EOjg values for the three halogenated biphenyls found to induce AHH (l.e., 3,4,3',4'-TC8; 3,4,5,3'T 5'-HC8; and 3,4,5,3' ,4' ,51-H88) were reported to average 4.5 nmoles per egg. In comoaring the potency of 3,4,3',4'-TCB with that of 2,3,7,8-TCOO and Aroclor 1254, 2,3,7,8-TCDO was almost 500 times as potent as 3,4,3',4'-TC8, and 2 x 10 times as potent as the commercial PCS mixture (124). 2,3,6,7-TC8P was found to be a very potent Inducer of AHH activity (EOjq 14 pmoles per egg), as was 2,3,6,7-tetrabromobiphenylene (TB8P) (EOjq - 22 pmoles per egg). Both of these blphenylenes were reported to be approximately equlpotent to 2,3,7,8TCOO and TCOF. 0C8P, 2,3,6,7-tetramethoxybtphenylene, and 2,7-dlnltroblphenylene (DN8P) could not Induce AHH activity at doses of 46 nmoles per egg (124). 2,3,6,7TCBP was also found to be roughly equlpotent to 2,3,7,8-TCOO in Inducing AHH activ ity in both mouse and rat livers. The ability of biphenyls and biphenylene congeners to compete for sterospecific binding of [^HJTCOO to mouse and rat liver cytosol was also investigated (124). 2.3.6.7- TC8P and 2,3,6,7-TB8P bound avidly, whereas 3,4,3',4*-TC8P proved to be approximately 30 times less potent. Table 6-1 compares the binding affinity In vitro. Kg (for mouse liver cytosol), and the biological potency in vivo, EOjg (for hepatic AHH activity in a chicken embryo), for 2,3,7,8-TCOO, several blphenylenes, and one biphenyl congener. As shown, 2.3.6.7- TCBP and 2,3,6,7-T8BP are roughly equlpotent to 2,3,7,8-TCOO. The other biphenylene analogs failed to conpete with [3H]TC00 for hepatic cytosol binding sites or for inducing AHH activity (_124). It should be noted that all of the biphenylene and biphenyl congeners which failed to induce AHH activity also did not exhibit any affinity for the cytosol binding sites. In contrast, the three halogenated blphenylenes and biphenyls which were found to induce AHH activity also com peted with [^HJTCOO for specific cytosol binding sites (124). HONS 215256 6-10 Tab Te 6-1 8I0LXICAL POTENCY AND BENOING AFFINITY OF VAR I DUS SIPHENYLENE AND BIPHENYL CDNGENERS (124) Compound TCDD 2,3,6,7-tetrachlorobiphenylene 2,3,6,7-tetrabromobiphenylene 3,4,3', A'-tetrachlorobiphenyl Octachtorobiphenylene 2,7-dlmtrobiphenylene Biological Potency i n Vivo 0.20 D.28 0.44 88 Inactive (470)* Inactive (930)* Blndlhg Affinity in Vitro (Kn fnMll 0.27 0.34 0.47 15 Inactive (540)* Inactive (540)* * The highest concentration tested that was judged to be soluble. Fish A considerable amount of extrapolation is required in order to interpret the availa ble data on fish, which pertains almost entirely to lethality (50). It appears that lethal levels of 2,3,7,8-TCOO In fish are within the same order of magnitude as those described for mammals (5). Rainbow trout that vere fed 2.3 mg/kg 2,3,7,8TCOD (representing a total dose of approximately 700 ug/kg body weight) exhibited a SO percent mortality rate within a 60-day period. Exposure of rainbow trout to 0.1 ug/1 of water over a 96-hour treatment period (representing a total dose of approxi mately 600 ug/kg body weight) resulted In 50 percent mortality within a period of 21 days (50). In comparative experiments with Coho salmon, a 96-hour exposure to an Initial water concentration of 0.105 U9/I (representing an estimated total dose of 5.4 ug/kg body weight) was found to be lethal within a period of 20 days (50). There was no significant Increase In egg mortality in either rainbow trout or pike when exposed to water levels as high as 0.01 ug/I over a 96-hour period (50). However, there was a significant Increase in the mortality rate of yolksac-fry in both of these species. MOMS 215257 6-11 An 80 percent mortality rate was observed m the feeding-fry of otke at an exoosure of 0.01 ug/1 of water over a 96-hour treatment period (50). There was no signifi cant difference in the same exposure regime for rainbow trout. Beported nistooatnological responses include edema, rupturing of blood vessels, and hecrotic livers. Teratogenic changes m embryos include shortened maxilla and structural defects of the operculum (50). - monkeys The most striking characteristic of animals exposed to dioxin is the substantial loss of body weight (109, 127). In addition to progressive weight loss, monkeys exhibit puffiness of the eyelids (IIP, 127, ^2^). Dryness and granularity of the skm, loss of hair, and reduced physical activity have also been observed (128). Two types of lesions have been reported m all laboratory animals studied: (1) involution of the thymus, and (2) testicular alterations, including atroohy, necro sis, and abnormal spermatocyte development (13). It Is believed that 2,3,7,8-TCDO has a suppressive effect on testicular microsomal AHH. Two other hyperplastic lesions in monkeys appear to be related to 2,3,7,8-TC00 or its metabolites: (I) bile duct hyperplasia, and (2) epithelial hyperplasia of the renal pelvis, urinary bladder, and ureter (IIP). Only one lesion, hypertrophic gastritis, has been observed In primates (221- This lesion is characterized by chronic inflammation of the gastric mucosa, which occurs in the fundlc and pyloric regions, conbined with small gastric ulcers penetrating the mucosa (33). In experiments with rhesus monkeys exposed to dioxins, researchers found reduced hematopoiesis, degeneration of blood vessels, focal necrosis of the liver, gastric ulcers, and reduction of spermatogenesis. Under gross observation, experimental monkeys exhibited obvious dilation of the heart, especially on the right side. Under microscopic examination, fibers of the cardiac muscle were distinctly seoarated by fluid, and Individual niscle cells tmre hypertrophic, with enlarged, dis torted, and typercftrotic nuclei. Although the lungs were not appreciably altered, isolated trees of atelectasis, congestion, edema, and fibrosis were observed. Their livers were small, firm, and moderately yellow, with many enlarged, multlnucleated parenchymal cells. Necrosis of parenchymal cells occurred In the centrilobular zone, and some areas of fibrosis were apparent In the periportal area. Their spleens tmre found to be small; the germinal centers tmre surrounded by only scat tered lymphocytes, and the blood sinuses were practically devoid of cells. The seminiferous tubules of the testes had abundant spermatogonia and sertpli cells. NOUS 213258 6-12 However, only a few primary spermatocytes Mre present, and no spermatids or mature spermatoioa were observed (33). The mesenteric lymph nodes of the monkeys were light tan and edematous. Microscooically, the nodes reseneled a splenic disarray of cellular architecture. Under gross observation, the bone marrow resembled coagulated plasma. Microscopically, only a few hematopoietic cells Mre seen within the marrow; these were equally divided be* t*en members of the myeloid and erythroid systems. Changes in the skeletal muscle resent)led those of cardiac muscle. Facial edema and petechiae Mr* commonly ob served (33). Whereas rats store 2,3,7,8-TC00 mainly in their livers (40 percent versus 10 percent in monkeys), monkeys accumulate the toxin in lipid tissues such as the skm, muscle, and fat (109). Consequently, unlike rats, the number of deaths attributable to liver insufficiency is minimal in monkeys (108) Rhesus monkeys that Mre fed 500 ppt 2,3,7,8-TCOO on a daily basis developed signs of toxicity at intake levels as low as 1 ug/kg body Might (112). A total intake of 3 ug/kg over a period of 9 months was sufficient to causa death (112, 127). The LD-50 of monkeys exposed to a single oral dose of 2,3,7,8-TCOO is approximately 50 to 70 ug/kg, with death occur ring wtthm 45 days (127). Prior to death, their peripheral blood is practically devoid of immature red and white blood cells (127) The marked throebocytopema was associated with widespread hemorrhaging (127). while death is usually associated wtth severe pancytopenia in chronically exposed animals, hematological changes do not always occur in those acutely exposed. Widespread hemorrnaging occurs only in chronically exposed animals (127). It appears that damage is greater to monkeys and other experimental animals when submilligram doses of toxin are administered over a period of time, rather than all at one time (1281. It has thus been speculated that toxicity is more closely related to the length of time that the compound Is able to interact than to the amount of toxin administered (127. 128). As exposure time lengthens, the severity of cellular deterioration in bone marrow and lymphoid tissue becomes widespread (127). The ob served ventricular dilation and myocardial hypertrophy may in part be related to the reduction In circulating erythrocytes and the subsequent increase in cardiac work load (127). Focal areas of hemorrhaoe were found in the epicardium, myocardium, and endocardium (127). Petechial hemorrhages were observed In the atria and the tips of the papillary muscles (127). HONS 215259 6-13 The possibility of reproductive abnormalities also exists. Altered levels of serum progesterone and estradiol, associated with both difficulties in conception and early abortions, have been reported m female monkeys exposed to low levels of 2.3.7.8- TCOO (127) Testicular atrophy may also develop in male monkeys given small amounts of dioxin (127) _ Considerable edematous fluid usually surrounds the epithelium of those exposed to 2.3.7.8- TCOO. Inflammatory cells, with a predominance of neutrophils, have been ob served to be in close proximity to ruptured submucosal mucinous cysts (127). The rupture of such cysts produces ulcers of the gastric mucosa (127). Metaplastic changes, characterized by numerous mucous -secret ing cells, have been observed m the ductal epithelium of the salivary glands, bile ducts, and pancreatic ducts. Similar changes may also occur in the bronchial epithelium and the palpebral conjunctivae (127). Hemorrhages within the brain have been limited to the Verchow-Sobbm's spaces surrounding the blood vessels (127). A series of single oral exposures were performed on female rhesus monkeys to es tablish a toxic (acute) dose for TCDF. The L0-50 value was determined to be 1,000 ug/kg, a dose approximately 20-fold higher than that estimated for 2,3,7,8-TCOD (129). Toxicity was characterized by the onset of the following symptoms 7 to 10 days after exposure (129): e Progressive weight loss, e Facial edema, e Loss of eyelashes and nails, e Development of white streaks in the eyelids, e Thickening of the skin, e Death within 2 to 4 weeks. The disease pattern cannot be distinguished from that produced by PCBs or 2,3,7,8TCODs (107, 128, 129). Oata pertaining to lethality, carcinogenicity, and reproductive effects are summar ized in Table 6-2 (50). HONS 215260 6-14 Tame 6-2 SIHMARY OF THE DOSAGE ASSOCIATED WITH THE TOXICOLOGICAL CRITERIA FOR VARIOUS PCOO CONGENERS HoaviOtt* \ttmr leiitt Utmilty Bditm to i.i.M-TCW SullWl C*rcmoi>*city Ojtt ibukHw l>tl ow a \ lllttv* til io ob*ve 2.3.7.a-'COO C^wt Pw y>%* lljCDO 2.7 2.a >2.000.000 300.000 >300.000 >150.000 33.000 33.000 TjCOO 2.3.7 30.000 10.000 15.000 >1.500 3.100 r4cn 2.3. 7,a l.j.i.a il..ij..si..ao 2-S.OOO <1.000.000 2 I ISO 1 2*0 1.700 33.000 1>10'J 1.10 X 8 i.t.j.x.a i.2..7, 1.2.3. a,7.0 i.2.3. a. 7.a 1.2.3.7.a.t 1.Z.3.S, 7,5 3-330 1.100-5.000 70-400 70-1300 40-1*00 1 1.100 73 70-100 40-100 1 5 170 540 >2.500 40 610 0 430 *0 >720 15 330.000 21 as 130 220.000 Mi 10'2 HyCOO l.2.3,a.t.7.l i.2.3..a.7. >400 >400 >300 370 10.000 OgCOO >1.000.000 53.000 HUMAN TOXICITY Chloracne, an often disfiguring and persistent dermatological disorder, is caused by human exposure to several chlorinated aromatic compounds, including PCBs. PCDFs, and PCODs (108. 130). This anomaly Is analogous to hyperkeratosis m mice, rats, raoblts, and monkeys (108, 130). Chloracnc may appear weeks, months, or even years after initial exposure to the toxin (3_3, 108), According to Moore (131), chloracne is the most sensitive Indicator of poisoning in the human subject. However, the absence of chloracne does not preclude Intoxication. Exposure to 2,3,7,8-TCDD may also cause polyneuropathy (nerve disease) and nystagimis (rapid eye movement). Liver dysfunction Is manifested by hepatomegaly and elevation of enzyme levels (33). Dally human exposure to 0.01 ug of 2,3,7,8-TCOO may result in incipient carcinogeni city, but existing evidence indicates that PCOOs are at most only mildly carcino genic (107). Exposure to 4 ug of 2.3,7,8-TCOO would result In a shortened lifespan, and dally exposure to 290 ug would likely cause acute toxicity (13). HOMS 215261 6-15 The result* of laboratory studies on 2,3,7,8-TCDD and its interaction with iron metabolism in rets (described in the preceding subsection) have significant implica tion for toxicity in humans. Persons with a high dietary iron intake would be ex pected to be more susceptible to 2,3,7,3-TCDD toxicity than persons with a marginal iron intake. Similarly, females might be less vulnerable to 2,3,7,8-TCDD toxicity than males, since they usually store less iron in their bodies (^3). This may ex plain the variation m toxicity levels among different species. The most notable cases of human exposure to 2,3,7,8-TCDO are those associated witn accidental releases in chemical factories or contact with contaminated materials/ areas (3_3), The immediate symptoms of exposure to dioxins include a burning sensa tion in the eyes, nose, and throat, followed by headaches, dizziness, nausea, and vomiting. Several days later, severe itching, redness, and swelling of the face (more marked over the eyelids, nose, and lips) may develop just prior to the onset of chloracne. It has been shown that as little as 20 ug of 2,3,7,8-TCDD on the skin can lead to chloracne f33). Within the initial weeks after exposure, inflamed nod ules as well as pustules appear on the face, forearms, shoulders, neck, and trunk, leading to comedones and cysts (33, 108, 130). Same times chloracne is preceded by erythematous and edematous skin lesions (108). After 1 or 2 months, acneform erup tions emerge, and the skin becomes hype augmented. At approximately the same time, mscles, primarily in the thl^is and chest area, becom aggravated upon exertion. Insomnia, extreme fatigue, nervousness, irritability, and the loss of libido also occur during this period (33, 108, 118, 130). Other symptoms of exposure Include arthralgias, porphyria cutanea tarda, hyperlipi demia, cutaneous hyperpigmentation, and hirsutism. Porphyria cutanea tarda, a skin condition that usually occurs as a photosensitive dermatosis, is caused by the de velopment of vesiculobullous lesions over exposed areas (J3). The dermatosis is pre cipitated by minor trauma, and may result in areas of healed bullae, crusts, scars, and ml 11 a (33). Blurred vision and the loss of taste and smell may alto occur (33). Patients with chloracne may exhibit Increases In serum triglycerides and choles terol. Uplds on the skin of such patients contain an increased proxrtlon of cho lesterol, sgualene, and wax esters. Since the latter two substances are known to induce comedo formation in experimental animals, it is possible that the disturbance of Hpid metabolism in cells of the sebaceous follicles is responsible for the der mal toxicity of 2,3,7,8-TCDD (116), HONS 215262 6-16 No fetal damage nas been substantiated to indicate a mutagenic, teratogenic, or embryotoxic effect of 2,3,7,8-TCDD exposure in vivo (33, 132). However, chromosomal abnormalities have been reported in in vitro cytogenetic studies on human lympho cytes exposed to 2,4,5-T containing D.D9 ppm 2,3,7,8-TCDD (33.). Chromatid breaks ana deletions increased with increasing concentrations of 2,4,5-T. It was not possible to determine whether this was a toxic or genetic effect (13). The clinical appearance of contaminated animals and the variety of let ions found during post-mortem examinations are identical for TCOFs, 2,3.7,8-TCDDS, and PC8s (1D7. 128). The toxicity of TCDF is approximately 10 percent that of 2.3,7,8-TCDD. and 1.D00 times that of Aroclor 1242 (128). The correlation of nmtabolism with de toxification indicates that the greater toxicity of 2,3,7,8-TCDD versus TCDF could be the result of the relatively slower metabolism and excretion of 2,3,7,8-TCDD (120. 133). Studies Indicate that TCDF nmtabolism is carried out through a detoxi fication nmchanism; species that are able to metabolize and purge TCDF are more re sistant to its acute toxicity. The multiple-organ and histologic dtMge may be a result of both the relative rate of formation and the further inactivation of the reactive metabolites in different organs (e.g.. liver, kidneys, stomach, etc.). Analyses of liver, adipose tissue, and blood samples of Yusho patients from both Japan and Taiwan indicated that the highest concentrations of PCOFs were found in the liver (Table 6*3). PC OF isomers present in the rice oil but not detected in the rusho patients were 2.3,6,7-tetra-. 2.3,4,6.7-penta-, 1,2.6,7.8-penta-, 1,2,3, 4,8penta-, and 1,2.3,4.6,7-hexa-COFs (Figure 6-2). These PCOF isomers, which were metabolized and excreted fairly rapidly, have two vicinal unchlorinated C-atoms in at least one of the two arometic rings In the PCOF nucleus (Figure 6-2) (79, 134). In contrast, PCOF isomers retained in the bodies, such as 2,3,6,8-tetra-, 2,3,7,8tetra-, 1,2,4,7.8-oenta-. 2.3,4,7,8-penta-. 1.2.3.7,8-penta-, 1,2,3,6,7,8-hexa-, and 1,2.3,4,7,8-hexa-CQFs, had lateral positions substituted with chlorine (Figure 6-2) (79. 134). The complete elimination of this compound from the organism is a fric tion of the body's ability to metabolize It to polar metabolites, which can be conjugated and excreted. MOMS 215263 6-17 Table 6-3 LEVELS OF PCOFs IN SAMPLES FROM YUSMO PATIENTS (79. 134) Liver___________ __________ Adipose____________ IsMer Yds ho, Japan YuSho, Tainan Yusho. Jaoan Vdsho. Taiwen 2,3,6,8-Tetra-COF <0-005.0.7 NO* <0.005-0.6 NO 2,3.7,8-Tetra-COF <0.005-0.3 0 060 <0.005-0.3 0.014 1,2,3,7,8-Pent4-CDF NO 0.194 NO 0.044 1,2.4,7,8-Penta-COF <0.005-7.1 0.042 <0.005-0.8 0.014 2,3,4,7,8-Penta-COF <0.005-6.9 0.091 <0.005-5.7 0.068 1,2,3,4,7.8-Me*4-C0F <0.005-2.6 0.193 <0.005-0.5 0.088 Stood Oitho. Japan Yusho, Taman NO NO <0 003 <0 03 NO 0 02-0 03 NO 0 04-0 06 0 003 0 08-0 120 <0.006 0.06-0 150 * H9/9. ** NO Not detected. 81-9 MOWS 215264 61-9 Atttietd E c r t* Figure 6-2. PCOf isown RcUined end Excreted in rusho Petients MOMS 215265 Section 7 CHEMICAL AND BIOASSAY ANALYSES CHEMICAL ANALYSES The toxicitles of son PCDO and PCDF isomers, which have bean demonstrated by tests (Section 6) and, to son* extent, by accidental exposures of huaians to these con* pounds (Section 2), have prompted extensive efforts to Quantitatively measure these chemicals in various chemical, biological, and environmental media (36. 126). These toxicity considerations have further dictated the requirement to analyte for these chemicals at concentrations ranging as low as parts-per-tr 11 lion (ID*** g or picogravs of analyte per grm of sample), development of an entirely new realm of ana lytical chemical capability has been required to achieve reliable quantitative de terminations at the part-per-trlllion level; this has only recently been success fully realized. Even when the sample medium contains higher concentrations of PCOOs/PCOFs, analysis for these cotpounds can be a formidable task because of the large nuebers and quantities of other chlorinated hydrocarbons which are frequently present in many samples of interest. These other hydrocarbons can complicate or even preclude accurate determination of the PCOOs/PCOFs If appropriate analytical methodology Is not utilized. In the present survey, the current status of analytical procedures for the determi nation of PCOOs/PCOFs Is reviewed. A summary of the Instrumental nethods which have been used for the detection and/or quantification of PCOOs/PCOFs is given in Table 7-1. While the focus of this survey is primerily aimed at techniques applicable to PCB-containtng fluids used in electrical devices, it is desirable to discuss the evolution of such analytical methods in a somewhat historical sense, particularly since most of the reported analytical procedures have been designed for other types of settle media. SCOPE OF THE ANALYTICAL PROBLEM As discussed earlier, there are 75 PCOO isomers and 135 PCDF Isomers. The magnitude of the analytical task associated with the determination of PCOOs/PCOFs in complex samples thus becomes apparent, especially when one recognizes the difficulty of separating and Identifying a large nuaber of Isomers within a given chlorinated 7-1 MOMS 215266 Table 7-1 INSTRUMENTAL METHODS APPLIED tO DETECT PCDDs/PCDFs in sample extracts tope 1eo',cit j" ScrMoioq/Xl*1 - It >1 4Q|'q1*i J Setcti-sicser "TliO-tl)*' TOO c*tt in* 31U 137 ill Eltctrgn-Ctaturi oil Chrgattafrignjr iC/K) *001 nr*ieiM 'sniMtignt. CfllgrMnang It, 1301 ',31, IK). *11. U! Satctrgaitry !nS I T*i*M*Hy 'dan14to* Coe- trw Cvtirt ill > tg^Mny ! *'*C3) *Hrsu of ml*.'i|h, (iglgsteil tlllu* 5ig1g*ic*l tlllu*!.'4t, TOO. -MntniliM. .grvig. 34if(4, gntntlitai. ooit'c'nt. TOD in* ml ituiurn 103 Iti ftcooo-CoIw lZ<kOo oHlutloo <S*)*ctM [oo Maiigno*) *ieoM-Coiio*i ec-Mifo t**g)utign *S i St loot** [go N*oi coring| ClO'l'irv-Co lu SC-oigo BtMlution *S tt.tctM Ion Mnitgri*) OMlttiry-Cclu* 30-*an BtHlutign *S I 5*lctM Ian ngmtgriny) CMlHirr-ColuMi C L1MI* CnrMts*PMnr IKIM CaluM SC-k< t*Miui1gn *S [S*l*ciM [gn MniUrinf) K >***ttir* Cumim 1 Igmuttgn ns en**tii* [gn ns 9C-AM*sgn*ri* ?-*tl*r* [gntutlgo 4 CM*r'nM sn*nH *300 *nttenlgrg*n*nl '330 **otacn|grg*n*nl - TOO nergtcim OriofO fgrnuHtign* - "COO Surfoct urn to* 'to) lirficn rcoo 1*5 '. 11? 111 i* l**Hf*cMeo.lMoHaoon* 11v*r TOO. mCOO. Ol**IC*l Hlttl. MU*Out *ff1u*0tl. Mill . -C30 Hoiaficii tuna, oitot nuniii -a l**f MIMS* TOO UlM*n nil, 300? * 10. outer. wgi**nt 'TO J*rtieu itti cmtigi laurcM * TOO 1 n iso 151 is? 131 IS? .5* StMritign gf '330. 'COD. oiCOO iM**ri. -*U)0. OcCOO. C3F. COOt lo fly itn C* 1" g*l. KH. *>r *m * ojrglyin grgtvctt. 'unto t'liu* *CS0* in* *C0FS 'ociogritgr fflutntl *CMl in* *COFs In OUM* tiisu* *cooiCMMitig* *ffiu*"t> . OtfMa IMlH C00l. tJl 1SS. .56. .31. .5?. 10 53. :s. 150 131 *9 '.so T9. 135 TOO. mCOO. MCOO. in* 0C30 ii**n m gtrticuiittt. <io**r iMirttign ii* iMntif'Cltign .3*. 1(1 Iniatl tiltM in* * *C00l nuatn nltot tutu*. r'r 1M *COO *C0F <" fH" M* iMIMnts 1 Motifleitig* of TCOO 'lain in ttinair* aiitir*. fit* '.S3. IS* 1*1 MOMS 7-2 class, ill of which tre quite similar structurally, and thus similar in chemical and physicil properties. This task is further complicated by the fact that mtny of the isomers of the PCDDs/PCQFs have not been prepared in pure form, and thus are not available for use in developing and calibrating analytical methods. Several synthetic routes for preparing PCDDs {.137., 16$, 166. .167, 168.~ 169. 170) and PCOFs {*5. 48, 68. 167, 171. 172. 173) hive been reoorted, and various PCOD and PCDF isomers nave been produced by these reactions. The entire sets of the isomers of tetra-, hexa-, and hecta-COOs, as well as octa-COO. have been synthesized (161. 169. 170), as have 2 di-, 4 tri-, and 14 penta-CDDs (^4, 174. C. Rappe. Personal Comnunication). All 38 tetra-COF isomers (175) and some of the penta-CDF isomers (174) nave been synthesized. The PCDO and PCDF isomers orepared by the various investigators cited have been characterized using several methods, including gas and liquid chromatography, mass spectrometry, proton- and 13C-IH1, and photolytlc dechlorination coupled with pat tern recognition techniques (136). However, few. If any (except 2,3.7,8-TCDD), have been structurally confirmed by single-crystal X-ray diffraction, which Is the most definitive method. This Is due in part to the fact that only quite small quantities of the synthesized isomers have been available; in addition, single-crystal X-ray diffraction requires considerable time and expense. The PCOD and PCDF Isomers prepared by these investigators have not been widely available to laboratories seeking these compounds for use as analytical standards. However, a limited number of PC00 and PCDF isomers, including several *3C- and 37C11 abelied compounds, have been prepared In sufficient quantities for sale to numerous investigators by a camamrcial synthetic laboratory (KOR Isotopes. Cambridge, Massa chusetts). PCDO standards are available only as mixtures; PCDF standards have been synthesized and isolated as individual Isomers. Even in those laboratories Fiich have synthesized or acquired PCOD and PCOF standards, there are no reported in stances in which the quantities of these cotpounds have been sufficient to thor oughly evaluate and calibrate the complex analytical procedures for entire sets of isomers of several chlorinated PCOO and PCOF classes. In spite of these limitations, there has been widespread Interest In the determina tion of specific PCOO and PCOF isomers. This Interest stems largely from the fact that the toxicity of these compounds is markedly dependent on both the nuxber and position of the chlorine substituents. HONS 219268 7-3 The toxicology of PCOOs and PCOFs Is discussed to re extensively in Section 6. It is clear fro* that discussion that there is a need to perform isomer-specific analyses of PCOOs and PCOFs In many types of sample media, and that simply determining these compounds by chlorinated class does not generally provide sufficient information to adequately evaluate hazards. However, the extent to which such-analyses can cur. rently be accomplished Is still rather limited for the reasons discussed above. Many of the early analytical studies focused on the determination of 2,3,7,8-TCDD m various biological and environmental samples, because of the extreme toxicity of 2,3.7,8-TCOO, which has been associated primarily with 2.4,5-trlchlorophenol and certain herbicide derivatives (2,4,5-T) of this chlorophenol (all widely used at one time), In addition, this PCOD isomer was one of the earliest available in suffi cient quantity to use as an analytical standard. Much of the currently extant ana lytical nmthodology for PCOOs and PCOFs thus stems from that developed originally to detect 2,3,7,8-TCOO. Some of this nmthodology was gradually adapted, modified, and expanded to accommodate analyses of other PCOOs and PCOFs, although entirely new net hods have also been developed. Among the many types of samples which have been analyzed for PCOOs and/or PCOFs are the following (36. 136): e Animal tissues - adipose, imiscle, liver, blood, whole animal (bovine, fish, deer, elk, turtle, rat, rabbit, birds). e Human tissues - blood, adipose, liver, kidney. e Human and animal milk. e Plant material. e Soils and sediments. e Hater - rivers, streams and lakes, wells, leachates. e Chemical formulations and wastes (2,4-0; 2,4,3-T, PC8s, chlorophenols, dlphenylether herbicides, and other herbicides and industrial chemicals). e ftafeustlon products - fly ash, bottom ash, soot, condensate particu lates and other stack effluents, water and other process fluids from codustlon of municipal waste, coal, wood, chemical wastes, PCBs, PCS-treated materials and wastes. e Industrial hygiene simples wipes, air samples (solid sorbents). e Hazardous waste site simples - complex chemical mixtures, landfill sludges, leachates. MONS 249269 7-4 It should be mntioned that the level of sophistication applied m these analyses, and the objectives thereof varied widely. In some cases, the objectives have been primarily to screen sanies (qualitatively or semiouantitatively) to identify chlor inated classes of PCODs and/or PCDFs. In other instances, the objective of the analysis has been to accurately quantify specific PCDD and/or PCDF isomers m such sanies. Both types of analyses can be useful, depending on the purposes for which the analytical results are to be used. The most specific of these Mthods employs mass spectronmtry (MS). Prereauisites for best analyses include representative sam pling, good storage, efficient extraction, and sample purification (containment) followed by good separation, ultrasensitive detection and quantification, and most desirable confirmation (1131. CRITERIA FOR SELECTING ANALYTICAL PROCEDURES There are several criteria which may be used to assess the efficacy of specific analytical procedures for the determination of PCODs/PCOFs, depending upon the ob jectives of the analysis. These criteria are discussed below. A standard practice applied by most competent laboratories idtlch perform analyses for PCODs/PCOFs Involves the addition of Isotopical ly labelled PCOO and/or PCDF isoamrs to the sample In Iuiomi quantity prior to analysis, and determination of these labelled compounds along with the native or unlabel led PCODs or PCDFs. The percent recovery of the Isotopical ly labelled standard provides a amasure of the efficacy of the analytical procedures. Typically, ^Cl-label led or ^C-label led PCODs or PCDFs have been used for this purpose. However, since only a few such standards have been prepared and are generally available (certain labelled isomers of the tetra-, hepta-, and octa-CDDs, and labelled 2,3,7,8-TCOF are commercially available), the extent to which these procedures can be utilized Is limited. The addition of a labelled PC00/PC0F to a sample Is also useful In that the added compound can be utilized as a true Internal standard for pirposes of quantitating the native PCODs/PCOFs m the sample. Such an approach presumes that losses of the added Internal standard In curred In the course of processing the sample are proportionately the sanrn as the losses of the native PCODs/PCOFs In the sample. Since the internal standard is added In an accurately known quantity, however, by measurinq the native compound relative to the labelled Internal standard, it Is possible to reliably quantitate the native compound In the sample. HONS 215270 7-5 Since PCDOs/PCDFs nave been detected at concentrations spanning a relatively *iae range in various sample matrices (typically oarts-oer-tnl lion to oarts-oerthousand), it is desirable to utilize analytical methodology uhicn can detect (and if desired. Quantitatively measure) PCODs/PCDFs over tms entire range. If large numbers of samples are to be analyzed, as is frequently the case m envi ronmental assessments, the analytical methodology should be amnable to efficient and reliable processing of such large numbers of samples in a reasonable tune per. iod. the complexity of reliable analytical procedures for determining PCDDs/PCDFs is such that even the best methods currently available are quite labor- and resource intensive. These analyses are therefore auite expensive, and single analysis often requires 1 to Z days, although economies can obviously be realized by orocessma samples in batches. To the extent possible, acceptable analytical procedures for determining PCDOs/PCDFs should be applicable to the analysis of a wide variety of different sample matrices. As shown in Tables 7-1 and 7-2, the procedures for separation and instrumental de tection which have been developed for PCDO/PCOF analyses are rather similar for many types of samples. SAMPLING tCTHODS The procedures utilized In obtaining a sample to be analyzed for PCDOs/PCDFs can markedly affect tne results obtained. A detailed discussion of sampling methods is beyond the scope of this review, but it should be noted that tms aspect of PCOD/ PCOF analysis is perhaps one of the most neglected areas. In general, tne dictates of good analytical practice should be applied In sampling: homogeneity of the sam ple matrix; collection of a truly representative sample; statistical considerations with respect to replicate analyses; and the resultant effects on precision and accuracy. In cases where samples are acquired by simple grab sampling, such consid erations can usually be applied. In other instances (e.g., collection of flue gas samples from incineration sources or surface wipe tests), the sampling procedures arc far less standardized, and their effectiveness and limitations have not been adequately evaluated. Sampling nwthodology is discussed In a recent Canadian Governnent reoort on analyti cal methodology applicable to PCDOs (185). A discussion of some of the sampling HONS 215271 r-6 procedures applicable for collection of flue oas samples has been presented oy `i(r. nan, et al. (160), and a detailed discussion of environmental samplmo reouirements and guidelines has been oreoared by the American Chemical Society 1136). 'able 7-2 list of EXTRACTION and CLEANUP PROCEDURES FREQUENTLY USED FOR PCDO/PCDF ANALYSES On oi nator U.S. Environmental Protection Agency U.S. Food and Oruo Admi nistration U.S. Fish and wild* life Service Dow Cnemical Company wnght State Univer sity-Srenm Laboratory Umea University, Sweden Procedure/Reference Acid extract ion (153) Neutral extraction (153) (176) (177) Acid extraction ( 178, 179) Neutral extraction (180) (180, 181) (160, 182) (10) (3, 183, 184) (79, 135) Apolleation Fish; adioose tissue; mi lx, water; soil; sediments Fish Chemical formulations, 'sn Fish Fish; milk Fly ash; dust, urban par ticulates; municipal sludae. SOI 1 Soi1. chemical waste: aqueous effluents Particulates and other samples from flue oas stacx PCBs Fly ash, soot, wipe tests Slood SAMPLE EXTRACTION, PURIFICATION, AND SEPARATION Analytical methods which have been developed for determination of PCOOs/PCOFs in various sample matrices generally entai1 a sequence of three operations: e Extraction of PCDO$/PCDfs from the sample matrix, which in some cases involves digestion or destruction of the matrix. MONS 215272 7-7  Preliminary separation of PCOOs/PCOFs from major matrix constituents and other major chlorinated residues and chemical contaminants. Quantification and confirmation of PCOOs/PCOFs in the c'eaned-uo sample extract. Procedures to accamolisn these ooerations have been develooed and applied m sever a' different laboratories. These procedures are similar in many respects, althougn they differ in various details, such as the choice of liquid chromatography columns, and the digestion or extraction reagents. A representatve listing of several metnods that are used by major laboratories currently performing PCDD and/or PCOF analy sis is presented In Aopendu 0. Other reported variations of such methods are dis cussed and evaluated m a recent Canadian reoort (^185h All of the methods listed m Appendix D entai 1 initial addition of internal stan dards, followed by destruction of the sample matrix (where practical) or extraction with an organic solvent, partitioning of the PCOOs/PCOFs Into an organic phase and polar constituents into an aqueous phase and discarding of the latter, and liauid chromatographic cleanup (and in some cases HPLC cleanup) of the extracts. The cleanup steps are intended to remove matrix components, such as lipids, as well as most of the chlorinated hydrocarbon residues in the sample (e.g., PCBs, DOE, other common chlorocarbon residues, and fused-ring aranatic compounds), ^iich may inter fere with subsequent detection and quantitation of PCOOs/PCOFs. Albro and Parker have recently discussed tne types of chlorinated hydrocarbons which may constitute interferences, and fractionation procedures aimed at elimination of these (_74)- Some of the more frequently used extraction and cleanup procedures are listed in Table 7 2, followed by a discussion of the efficacy of extraction and cleanup rnetnodoloay. EFFICACY OF EXTRACTION AND CLEANUP METHOOOIOGY FOR PCOOs/PCOFs The efficacy of extraction and cleanup nsthodology for determining PCODs/PCDFs is generally gauged by the extent of recovery of the initially added labelled internal standards. This has been applied less frequently for PCOF determinations, because only one labelled PCOF has been available (^7Cl4-2,3,7,8-TC0F). Collaborative test ing of some of these methods has been accomplished through EPA's Oloxin Implementa tion Plan (180). Few of the results of these studies have yet been oubllshed. The FQA also conducted a limited assessment of several of these nmthods for fish analy ses (187). Most of the methods cited seem to be relatively effective m so far as they have been tested, when accomplished by skilled and experienced analysts. MEWS 215273 7-8 One type of simple which has received particular attention m terns of evaluating the efficiencies of extracting PCDOs/PCDFs using various orocedures is oarticulates (such as fly ash). lustenhouwer. OUe, and Hutzinger (60) have resorted the results of extracting aliquots of the same fly asn sample using six different extraction methods, which have been used by various investigators who nave reoorted data on de terminations of PCDOs/PCDFs in the literature. According to these results t'"able 7 3), Soxhlet extraction with toluene (following acid treatment) was the most effec tive of the several procedures evaluated. More detailed subsequent studies from the same laboratory in which still other oro cedures were evaluated led to the conclusion that Soxhlet extraction with benzene 's approximately equally effective, and far superior to simpler extraction metnoos (378). In the latter study, the effect of the period of extraction, ranging from 24 to 72 hours, was also evaluated. The results of these tests, shown m Table 7-4. indicate that a 24-hour extraction period was sufficient to remove the bulk of the PCOD/PCDF constituents. Cutie (153) has also reported results of similar studies on the extraction of 2,3,7,8-TCDO from fly ash, soil, and active carbon. There have been comparatively few measurements of PCDDs/PCDFs m electrical eauipment PCS fluids (see Section 3). As such, methodology for this purpose has not been extensively reported. The extraction and cleanup procedures applied m several of these studies were rnerally similar to those described earlier in this section for determinations of PCDOs/PCOFs m other samples. The most detailed accounts of such orocedures appli cable to PCB electrical fluids are those given by Suser (166). Happe. et al (371. and Albro and Parker (7). Polychlorinated diphenylethers, which are also present in some fluids of this type, can give rise to mass spectral fragment ions, which coincide with those monitored as indicators of the COFs (_71. It is thus important that the chlorinated diphenyl ethers be effectively removed from the extracts by sep aration techniques prior to GC/MS analysis. Adeouate separations can be achieved by careful attention to the elution procedures used in the liquid chromatographic frac tionation sequence (74). INSTRUFCNTAL CTHOOS Table 7-1 lists the instrumental methods which have been applied for detection and/ or quantification of PCDDs/PCDFs in sample extracts prepared by the orocedures given in Table 7-2 and Appendix 0. Table 7-1 shows the types of analytical applications HONS 219274 7-9 Tib le 7-3 EXTRACTION EFFICIENCIES OF POLYCHLORINATED COMPOUNDS "POM FLY ASH (60) Analytical Results f"otal. sop) Method 1. Toluene, acid treated, Soxhlet extraction Chi orooheno 1 PCOD - PCDF 1364 10 1309 1001 1407 1 OOI 2, CHjClj, shaking treatment 240 18 3. Acetone/hexane, ultrasonic treatment 92 7 124 1 23.5 2 NO 1 NO 1 A. CHjClj, Soxhlet extraction 5. CHClj, Soxhlet extraction 434 32 592 44 81.8 168 6 13 155 11 260 20 6. Toluene extraction of solution of fly ash in 4 HF 446 33 113 9 230 16 Description of methods: 1. Fly ash (25 g) is stirred In In hydrochloric acid (200 ml). After centrifu gation, the residual fly ash is washed with deionized water over a Buchner funnel and dried at room temperature. The fly ash is extracted for 35 hours ,, with toluene using a Soxhlet apparatus. 2,3. Fly ash (10 g) is shaken manually (Method 2) or treated in an ultrasonic bath for 5 minutes (Method 3) m the solvent indicated. For ooth "etnods, this is reoeated four times. The extracts are combined for cleanup and analysis. a,5. Fly ash (25 g) is extracted for 35 hours, using a Soxhlet apparatus. 6. Fly ash(5 g) Is dissolved in a &8X HF solution. i>e solution is then extracted with toluene. Some particulate matter renamed. MONS 215275 7-10 (e.g., screening or qualitative detection, quantitative analysis, isomer determina tions) for nich these various methods nave been aoolied, alona with examples of reoorts of such aoplications. As indicated m Table 7-1, several methods nave seen developed for rapid screening of samples for the presence of PCODs/PCDFs, sucn as DV-soectroscoPy. thin-layer chromatooraphy For example, gas cnromatoorapny jsino an electron capture detector (GC/ECO) offers the sensitivity needed for such a screening procedure, but does not have the specificity that is needed" td eliminate false positives. The ECO responds to any molecule that has an affinity for captur ing electrons m an ionized field. This includes molecules containino halogens (e.g., pesticides), oxygen (e g,, phthalate esters, ethers, ketones, etc.), and dther hetero atoms, such as sulfur and nitrogen (144). Table 7-4 INFLUENCE OF TIME ON EXTRACTION EFFICIENCY FOR PCODs AND PCOFs* (188) 1st 24 hours 2nd 24 hours 3rd 24 hours 1st 24 hours 2nd 24 hours 3rd 24 hours TCOO 97.0 2.3 0.7 P^ 97.4 2,0 0,6 PCDD ftl HCOD HtCDO 98,4 1.3 0,3 99.3 0,7 0,1 OCDO Average 99. S o.s 0.0 98,0 1.6 0,3 TCDF 97,7 1.7 0.8 ft 98.4 1,3 0.4 PCOF <D HCOF htC0F 98.6 1,1 0.3 99,4 0.6 0,1 OCOF Averaoe 99,5 0.5 0,0 98,5 1,2 0.3 * Sample pretreetnnt with IN NCI; Soxhlet extraction with toluene. Radian Corporation has developed a prototype detector system for gas chromatography, called a thermally modulated electron capture detector (THECD) system. This system consists of two electron capture detectors in series with a temperature-contro! led HONS 215276 7-11 catalytic pyrolysis unit between them. Different classes of organic conuounas react to the pyrolysis conditions m different ways, resulting either in an enhanced sig nal, a diminished signal, or no change in the magnitude of the signal The ratio of the peak area measured for the detector after the reactor to that for the detector prior to the reactor is a property cnaractenstic of each eomoound This oea* area ratio is used m conjunction with its retention time to increase the confidence level of the identity of a oiven compound - In addition to this methoa (which is still unoer development and needs validation), several techniques have been developed which are dependent upon biological responses to the toxic PCODs/ PCOFs. These bioassay techmoues are discussed separately m this section. It is generally accepted that the only technique which has both the required sensi tivity and specificity for reliable detection and quantitation of PCDDs/PCOFs is coupled aas chromatography-mass spectrometry (GC/MS). Hass spectranetnc analysis for PCDOs/PCOFs is based on the presumption that these compounds exhibit specific mass spectral fragmentation patterns. The electron impact mass spectra of PCOOs and PCOFs exhibit Intense molecular ions (M+) with the appropriate ion clustering, which is due to the two chlorine isotooes. PCODs mainly fragment to M+-C0C1 and M+-2C0C1 ions. Smaller intensities of various other ions aopear in the spectra of both types of standards, thus enabling reliable analysis of PCDDs/PCOFs (139). In most instances, monitoring the molecular ions of the PCDOs and PCOFs will easily distinguish these compounds from otner chlorinated aromatic nyarocarbons, which may be present In the saavle extracts. However, some PCBs, as well as DDE, can fragment to yield ion masses which may interfere with the detection of certain PCOOs, unless rather high mass spectral resolution Is employed. Also, the identification of PCOFs may be complicated by the presence of chlorinated diphenylethers, since tne latter compounds yield intense M^-Clj Ions, which have the sant exact mass and nuntoer of chlorine atoms as the corresponding PCDF (156). In such cases, successful analysis is dependent upen the ability of the preseparation procedures and/or the gas chroma tographic fractionation to separata such interfering compounds from the PCDOs/PCOFs before the latter enters the mass spectronpter. while it is desirable to obtain complete mass spectral scans in GC/MS analysis in order to identify tne analytes of interest, this is not possible If very low detection limits (e.g., part-per-tri 1Hon) are needed. In the latter case, which is the more comnon requirement m ana lyzing environmental simples, only a few ions in the mass spectra of the PCODs/PCDFs MOMS 215277 7-12 of interest are typical ly monitored, utilizing a technique termed Selected-Ion Momtor mg. This is discussed further below. Earlier analyses of PCDOs were aimed primarily at determining the low levels of TCDD, and utilized less soomstieated gas cnromatograpnic techniques than those presently available. The first attempt to apply mass spectronetry for quantitative determination of TCDD was the high-resolution mass soectromttnc technique reoortea by Baughman and Heselson {1*31. This technique did not utilize prior us chromato graphic separation at all. the sample extract was introduced via a direct inlet probe. These investigators modified a high-resolution mass spectrometer to permit repetitive high-resolution scanning over a narrow mass range (typically 0.3 AW) at a rapid rate (4 scans/second). Hass spectranetric signals were acquired using a signal-averaging computer, which resulted in sufficient sensitivity (approximately 1 picogram of injected TCDD). Due to the relatively high mass spectral resolution achieved, this technique was adequate for the purposes of distinguishing the molecular ion of TCDO from ions arising from the sample matrix (mostly biological samples). However, it was soon realized that the TCOO ions could not be distinguished from those originating from certain PCBS, if the latter were present in the sample extract in large concentra tions relative to the TCOO (part-oer-milllon versus part-per-trilllon). Another limitation of the latter method was its inability to separate TCOO isomers; the mass spectrometric response observed with this nmthod was obviously attributable to any TCOO isomers in the sample, or any compounds given a TCOO fragnpnt, not just the 2,3,7,8-TCDD which was of major interest. During the san period that the Baughman-Meselson method was being applied, several investigators reported the use of coupled gas chromatography-low resolution mass spectrometry for deteneinations of TCOQs m chlorinated phenols and 2,4,5-T herbi cide formulations (145-149). These studies utilized open-tubular packed gas chroma tographic colwmis, which were capable of separating TCOO from other PCDOs of differ, ent chlorine content, but had little capacity for separating discrete TCDD isomers. The detection limits achieved in these determinations were in the range of 0.001 to 0.1 ppm. These methods derived their specificity from the cleanup methods used. Furthermore, these wthods required that the analyte exhibit the appropriate GC re tention time and a mss spectral response at the appropriate Ion masses corresponoi ng to TCOO. HONS 215278 7-13 Due to concerns over the possible accumulation of 2.3,7,8-TCDD in the food chain (from the use of 2,*,5-T on cattle-grazing land), the U.S. EPA initiated the Dioxin Implementation Plan (DIP} in 197S. This program was intended to Quantitatively assess the levels of 2,3,7,8-TCDD in beef tissues and related environmental samples The program prompted the development of conbined GC (packed-eoluwi)-mgh resolution mass spectrometric techniques for determining TCDD at four different laboratories i.e., Wright State University, University of Nebraska, EPA/RTP, and Dow Chemical Company (136, 150. 152. IBlI The mass soectrometnc methods were similar to meth ods applied by Baughman and Meselson (143). but were superior m that hlqh-resolution scanning was generally accomplished over a narrovmr mass range and at a more rapid scan rate. This improved both the resolution capability and the sensitivity of the technique. Wright State's adaptation of this procedure permitted as many as four ions to be monitored. A further enhancenent of the capabilities of the GC-hlgh resolution mass soectronetrlc technique for analysis of PCDDs/PCDFs entailed the incorporation of capillary GC coliums (wall-coated open-tubular columns). This has been accomplished at both the EPA/RTP (153) and Wright State laboratories (ISA). The use of caoillary GC col onels provided the capability to separate discrete isomers of the PCDDs/PCDFs; until recently, few of these have been available in pure form. The use of caoillary col umn GC-hlgh resolution MS also further improved the ability to obtain accurate quan titative data on the low concentrations of PCDDs/PCDFs in environmental samples. Typical results of this highly specific technique are showi in Table 7-5. Due to patent problems in the United States, the technique of capillary columns was developed and extensively used In Europe. Investigations of the use of capillary GC columns for the separation of PCOO Isomers *re undertaken as early as 1975 by Buser (155). who prepared eight HxCOD Isomers and demonstrated that these could be separ ated on a column having about 35,000 theoretical plates, using a temperature program which Increased the coluxei temperature by 2*C/m1nute. Subsequently, Buser (155) demonstrated separations of nine TCDD Isoners available at that time. More re cently, Outer (155) and Outer and Rappe (157) have reported the results of separat ing all 22 TCDD Isomers on three different capillary coluneis. Other capillary coluneis have also been tested for separating the 22 TCOO Isomers, such as new hybrid coluxms coated with stationary phases OV-17 and/or STlar-lOC (Figures C-l and C-2, Appendix C). It should be noted that these techniques pro vided for the separation of the highly toxic 2,3,7,8-tetra-COO isoner from the other HONS 215279 7-14 Table 7-5 typical analytical results for quality assifance SAMPLES. ANALYSIS OF FISH FDR 2,3,7,3-TCDO (153) Sample* (g> 5{ 1)* 5(1) 5(1) 5(4) 5(3) 5(3) 5(3) 5(1) 5(1) 10(1) 10(1) 10(2) 10(2) 10(1) 10(4) 10(1) 10(1) 37C1-TC00 Recovery** EX) 62 52 82 100 54 100 78 92 97 100* 100* 100* 100* 100* 67 93 84 TCDO Detection Limi t (opt )* 2 4 3 3 1 2 2 2 5 1 4 7 3 4 1 3 4 TCDD Detected (opt)* 20 34 NO NO 19 NO NO 19 45 8 43 NO NO 76 NO 56 73 TCDD Fortification Level P9 001 no 22 185 35 00 D0 70 14 00 00 55 11 240 48 130 13 600 60 00 00 1250 125 00 650 65 620 62 * (1) Ocean perch, (2) lake trout, (3) beef liver, and (4) method blank. * Each sample has been fortified with 5 or 10 np 37C1-TCDD. # Corrected for X recovery losses; NO not detected; 37C1-TCDD mean X recovery, 36X; TCDO mean X accuracy, tl5X. HONS 215280 7-15 21 isomers. Such capillary colunrn also provided the means for separating oentaand hepta*C00 isomers (Figure C-3), as veil as tri-t tetra-, oenta-, and hexa-CDFs (Figures C-4, C-5, and C-6). Capillary colum GC-!ow resolution mess soectrgnetry is being increasingly utilized at several laboratories to provide data on a wide range of PCDD/PCOF isomers in var. ious types of samoles (_10, 3. 136, 1S5. 157. 158, 159) including PCS "electrical fluids. A summary of the Quantification of separate PCOD and PCOF isomers in sam ples from PCB fires and incinerator fly ash is oiven m Table 7-6. Dow Chemical scientists have recently achieved considerable success in the seoaration of tetra- and hexa-CDO isomers by HPLC fractionation of discrete isomeric com pounds (136, 161). This technique appears to be particularly powrful when used m conjunction with GC/MS techniques, and does not necessitate the use of capillary GC columns, ho data on separation of PCOF isomrs by this method has been reported. Several newer mass spectrometric techniques have also been recently investigated for the analysis of PCDDs/PCOFs. Among these are negative-ion mass spectrometry (163). negative-ion chemical ionization mass spectrometry (162). and atmospheric pressureionization mass spectrometry (142). Results obtained thus far show that all of these offer enhanced sensitivity, selectivity, and specificity. For furan isomers in particular, there is a dramatic increase in sensitivity using negative-ion chemi cal ionization NS as coaoared to normal electron impact NS. quantification To date, quantification has been based on peak area measurements by comparison with either internal standards or a calibration curve of known quantities of authentic standards (external standards). It has generally been assumed that all isomers of a particular PCOD or PCOF have the same response using the NS quantification. Re cently, Gara, et al. (189). and Rappe, et al. (113), have studied responses of 28 and 20 different PCOF isomers, respectively. Findings of both studies shoved a 3* fold variation in the response, using the electron impact (El) mode, and uo to a 20 fold and 25>fo1d variation, respectively, using the negative chemical ionization (NCI) mode. For the higher chlorinated homologues, the differences appeared to be much smaller. HONS 21S2S1 7-16 a -i Tab It 7-6 QUANTITATIVE ANALTSES OF PCOF AMO PCDO ISOPCRS IN SAMPLES FROM PCS INCIDENTS AMO INCINERATOR Fir ASH Isoaer Mono-CDF Oi -CDF Tr i-CDF Telra-CDF Penta-CDF Heia-CDF Nepta-COF Octa-CDF II nan aton* () - - 21 670 MS 460 40 Nwrtal Je** (9/9) 20-4] 15-43 5-14 3-4 Sfcovde# (ua/e7) - - 0 01-0 6 01 0.06 0 DOS 0.005 SurahaBaarf - - <0 02-4.0 <0.01-] ] <0 01-1 0 <0.015-1 5 0.002-0.3 Municipal Fly Ash Ill 196 361 177 18 Part leu)ate __ - * 460 960 1600 1130 140 Inttistnal Fly Ash** (PI* I - - 110 530 ]S0 300 20 Municip41 F ly Ash _ippt>) - - 9 23 50 2S0 ns Tetra-COO Penta-COO mu-COO mpta-COD Octa-COO 1.2 s.o 4,7 70 2.0 154 100 182 800 326 1370 288 1370 106 310 400 1400 1000 300 ISO 3 20 50 180 210 * Reference 29. **-Reference 2. I Reference 190. t Reference 60. MOMS 215282 COff IRHATION Confirmation of PCOOs/PCOFs can tie based on the prxer to Secular ions and frjon*nts, as well as the intensities of the ion clusters due to the Cl lsotooes. Moreover, a samole contains a multitude of PCDD/PCOF isomers, the positive identification of PCDOs/PCOFs could be confirmed by cluster analyses (113). For adeouate confirmation of PCDDs/PCDFs, there are several oaraneters available Cornered to an authentic standard, the PCDOs/PCDFs in the SMole should have tne same following parameters: retention volume m tne cotum chromatography m the cleanup steps, retention tine m the gas chromatographic separation, molecular weight, fr ague n tat ion and chlorine cluster in the Ms detection, and difference m response factors comparing the El and the NCI mode. These paraneters, in addition to access to welt defined weighted standards, dive the possibility of reliable anal ysis of PCDOs/PCOFs. BIOASSAYS Numerous cllnlctl biomedical parameters have been shown to be altered in animals and humans receiving different doses of PCDOs, PCOFs, and PCBPs (Section 6). In a num ber of investigations, 2,3,7,8-TCOO served as a prototype of individual PCOOs and PCOF congeners end their mixtures, alt of which produce a similar and characteristic pattern of toxic lesions, including hyperkeratosis and souamous metaplasia in the skin, and induction of a nuafeer of coordinated expressed enzymes through stereosoeciflc, reversible binding to a cytosolic receptor protein (which has been identified m several mammalian soedes and In celt cultures). As described in Section 6, the potencies of the bioloaic and toxic responses for halogenated aryl hydrocarbons are also dependent on structure. Maximal activity was observed for the 2,3,7,8-TCOO congener, which contains a chlorine substitution pat tern at the four lateral positions of the dlbenzo-o-dloxin molecule (igi). Further Cl substituted at the 1, 4, 6, and g positions tends to diminish the activity, whereas the removal of one or acre of the laterally substituted Cl atoms leads to a marked loss (or complete loss) of activity. The results of recent studies suogest that the initial event in the mechanism of action of 2,3,7,8-TCOD and related com pounds is the reversible non-covalent binding to a receptor protein. There appears to be an excellent correlation between the relative toxicities of halogenated ary] hydrocarbons and their potencies as AHH Inducers and their receptor protein binding avidities (_18l). This suggests that both the receptor binding and AHH induction MOMS 215263 7-18 assays may be useful short-term in vitro bioassays for total PCDO/PCOF biological activity, which would in turn reflect potential toxic potency. The significance of the bioassays, in addition to the assessment of the toxicologi cal effects, is their usefulness as a raoid and reliable screen for detecting biolo gically active substances pnor to chemical identification and quantification. The following discussion will focus on some of these bioassay techniques. Aryl Hydrocarbon Hydroxylase (AHH) Induction 2,3,7,8-TCOO has been known to alter the activity of a number of enzym systems, the most extensively studied being the hepatic microsomal monooxygenases. The monooxy genase enzyme system, consisting of KAO PH -cytochrome P-450 reductase and cytochrome P-450, is an integral part of the smooth endoplasmic reticulum, and catalyzes the catabolism of lipophilic caepounds to more polar mtabolites. The induction of the hepatic monooxygenases by 2,3,7,8-TCOO occurs in most tissues of most species. It should be noted that these bioassays are extremely sensitive to a variety of chlorinated aroawtlc hydrocarbons. A large number of specific enzymes have been assayed; the most extensively studied of them, AHH, is discussed below. AHH activity is a measure of one cytochrome, cytochrome P-448. The induction of AHH is mediated by the reversible stereospecific binding of halogenated aroxatle hydro carbons to a cytosol protein, the cytosol Induction receptor (192), which mediates the nuclear uptake of the toxicant (193). The receptor Is determined by the Ah locus in mice (192). The Ah locus, and hence the cytosol receptor, controls not only the induction of AHH activity but also a battery of coordlnately expressed enzymes (192). Two lines of evidence suggest that toxic lesions produced by TCOO, as well as bio chemical responses, are mediated by the cytosolic receptor. First, the binding affinities of a large nuafcer of halogenated aromatic hydrocarbons for the receptor correspond very closely with the rank order of their toxic potencies (193). Second, two toxic responses produced by TCOO in mice, thyenc involution and cleft palate formation In fetuses, have been shown to segregate with the Ah locus (192). Caaoared with classical polycyclic aromatic hydrocarbon Inducers of AHH, such as 3-methyicholanthrene, 2,3,7,8-TCOO produces a parallel dose-response curve; both MONS 215284 7-19 coaoounds produce the same maximum response Tn AHH activity. However, 2,3.7,8-TCDO is 3 x 10* time* more potent that 3-methyl cho lanthrene as an AHH inducer. Poland and coworkers have extensively studied the induction of AHH and other enzynes by 2,3.7,8-TCDO and a variety of its coroners (50). This structure-activity rela tionship has been examined using a variety of PCDD (SO) and PCOF {_5, 197) congen ers. The AHH induction bioasssay utilizes a rat hepatoma cell line which possesses low basal AHH activity and Is highly inducible for this enzyme. The amount of ahh induction is determined by the fluormetric assay of 3-hydroxybenzo[a]pyrene forma tion from benzo[a]p>rene (the substrate for the AHH enzyme assay), or by the fluorimetnc determination of 7^thoxyresonif1n 0-deethylatlon. This technioue has been used to examine Individual halogenated aromatics, food, and environmental extracts (191). For examole, several gelatin samples have been shown to contain pentachlorophenol and related PC00 imuritles, including the 1,2,3,6,7,8- and 1,2,3.7,8,9-hexaand 1,2,3,4,5,7,8-heota-CODs which exhibit AHH activity. The results confirmed the correlation between the concentration of the toxic dioxins In the samples and their bioassay 2,3,7,8-TCOO equivalents. The binding of a specific ligand or mixture of ligands is determined by measuring the dose-dependent displacement of [3h]-2,3,7,8-TC00 previously bound to the pro tein. Toxicity to Cells Cultured in vitro One of the most characteristic responses to TC00 in vivo is hyperkeratosis in the skin. Recent work by Knutson and Poland has demonstrated that the hyperkeratlnizatlon rtsponse to TCDD exposure which occurs upon exposure to TCOD in vivo could be reproduced In an In vitro model system (192, 195). When XB^olls derived from a mouse teratoma are cultured at high density (to prevent spontaneous dlffentlatlon) along with lethal ly Irradiated 3T3 cells, the addition of TCOD produces a dose-related keratlnlzatlon response, as detected by red staining with Rhodanlle blue. This Is a direct effect of TC00 on XB cells, and will occur m the absence of the feeder cells If the teratoma cells are cultured in 3T3-condi tloned media. XB cells contain the cytosol receptor, and respond to TCOD with a dose-related Induction of AHH activity. HONS 2152B5 Other hatogenated aromatic hydrocarbon congeners, including PCOOs, PCDFs, PCBPs, and azobenzenes, also induced keratin nation although to a lesser extent than 2,3,7,8TCOO. Keratlnizatlon could also be induced by some nonhalogenated aromatic hydro carbons, such as benzanthracene and 5,6-benzoflavone (192). Since 2,3,7,8-TCDO is the most potent, but not the only inducer of this hyperkeratmizatlon response, tnis effect Is referred to as "dioxin-like" activity (196). unrelated toxins were studied to determine if the keratinlzation response was due to nonspecific cell dam age (192). The direct acting alkylating agents N-acetoxy-2-acetylaminpf luorene and N-methyl-it'-nitrosoguanldine; inhibitors of nucleic acid synthesis, actinomyem D, bleomycin, and cordyccpln; the spindle microtubule inhibitor, colchicine; kepone, a chlorinated hydrocarbon producing a toxic syndrome distinct from that of TCOO; and 2,6-dichlorobenzonitrlle, an acnegen considered to have a different mechanism than TCOD all failed to produce keratlnizatlon in the XB/3T3 culture system (192). Fur thermore, a correlation has been demonstrated between the de^ee of toxicity of var ious TCOD isomers and congeners and the extent of hyperkeratlnliatlon response (124. 192). The XB/3T3 system is extremely sensitive, qivlno an Induction of keratlnizatlon with as little as 3-2 plcograae of TCOD (192). In addition, this system Is particularly relevant as a model, since it deals with an Induced differentiation of the intact cell using an endpoint (hyperkeratlnliatlon) known to be relevant to human exposure (l.e., chloracne) (196). MONS 2152B6 7-21 Section s RESEARCH NEEOS INTRODUCTION This section outlines research recommendations directly related to the potential formation of PCOFs, PCODs, and other polychlorinated aromatics from askarel fluids. These recommendations, based on the findings presented In previous sections, reflect the identified Information gaps regarding the generation of these compounds, them toxicity, methods of analysis, environmental pathways, and procedures for their de struction /remove I. GENERATION PCOFs, PCDDs, PCBPs, and other polychlorinated aromatic compounds have been gener ated in association with PCB equipment fires and explosions. The quantities gener ated were found to vary significantly, depending on the type of equipment and reac tion conditions. The probability of generation under normal operating conditions and in noncatastrophic malfunctions Is still highly uncertain, Since available information on both conditions and quantities of formation is insuf ficient and often contradictory, future studies should focus on better delineating the generation of PCOFs, PCODs, and PCBPs. In parallel, standard and non-standard operating conditions should be compared with optimum formation conditions to deter mine (1) if formation is Indeed possible, and {2} if the necessary precursors exist in PCB fluids. Operating conditions of primary Interest Include the fotlming: e long-term high temperature overload in the presence of an oxygen source {e.g., water, o2). Short-term catastrophic failure caused by internal arcing, o long-term part i al di sch arge. e Arcing in circuit breakers containing PCB-contaminated oil. By conducting the above tests under controlled conditions, performing the analyses using the most advanced analytical techniques, and comparing the resulting data to MONS 21S2B7 8-1 established theories, the above expeniwnts would provide specific insights into the foltowing: e Reaction conditions favoring the formation of PCOFs from PCS*. e Mechanisms and wtimum reaction conditions for the formation of PCODs from chloroohenols and chlorobenzenes. - e Chemistry and reaction conditions for the formation of PCfPs. e Effects of dilution of PCBs m mineral oil on the formation of PCODs, PCOFs, and other cnlorinated species. TOXICITY Environmental contaminants become a concern when they affect human safety and the duality of the environment. It Is therefore Important to assess the risks resulting from exposure to the above expounds using the appropriate toxicological criteria. Milte other PCDO homo logues have been studied to a lesser degree, the data on structure-blologlcat activity relationships for 2,3,7,8-TCOO In laboratory animats are extensive. These data pertain to lethality, carcinogenicity, and reproductive ef fects. Toxicological date for PCOFs and PCSPs are very limited, as are the data on the syn ergistic effects of two or mere of the above by-products. For these compounds, spe cific data are lacking on acute lethality, chronic toxicity, carclnogenlclty, tera togenic and reproductive effects, mutogenlclty, and Immunotoxlclty. There are sig nificant gaps In the relationship between the receptor theory and the Induction of chlorecne. Data on such effects as the extent of enzyme Induction through specific receptor binding and cell keretlnlzatlon should be acquired through bioassay teennlques. In particular, rmseardi should be focused on the human target of PCODs and PCOFs in at leut three areas: Dotage/time/response relationship for each major health effect actu ally seen In humans as a function of exposure to potential PCODs and PCCFs. e Mechanism of action at the tissue level for each major health ef fect of these chemicals In hueans. e Quantitative risk assessments for each real, ehronic health effect In humens under exposure conditions actually met In occupational or envlromentat situations, ' MOMS 215268 8-2 CHEMICAL ANO BIQASSAY ANALYSES It it apparent that there are several areas of research which recuire additional em phasis in order to further advance the capabilities for analysis of PCOOs/PCDFj. Such areas include the following: e Synthesis and isolation of additional pure PC00/PC0F isomer stan dards for use in calibration and metnods development. ' e Synthesis of additional compounds which are chemically similar to PCODs/ PCOFs, and which may represent interferences In PCOO/PCOF analysis (dlphenylethers, hydroxylated- and mmthoxy-OPEs and PCIs) for methodology evaluation. e Development of htgher resolution GC and/or iC columns for Isolation of PCOO/PCDF Isomers. e Research on more specific detection, quantification, and confirma tion methods, including possibly positive and negative ion chemical Ionization MS, atmospheric pressure ionization MS, and MS/NS. e Research Into an Isomer-specif 1c analytical method to differentiate between toxic and nontoxic isomers. e Research Into rapid chemical and/or biological screening metnods. e Standardization of sampling methods, particularly for such tech niques as wipe tests, and establishment of criteria to Identify the significance of analytical results obtained via different samollng methods (e.g., how the results of a wipe test relate to toxicologi cal deta). ENVIROfMEKTAL persistence The environmental pathways of these camounds need to be better defined. Neither the pathways nor the rates (other than general overall rates) for their physical mi gration and decay have been determined with any certainty. Photolysis, for example, has been called the main route of environmental degradation. However, this process requires a hydrogen donor, rfilch Is not likely to be present under most normal envi ronmental conditions. Thus, further research Is needed Into photolysis to define the role and afcmndaace of hydrogen donors, and the exact conditions under which measur able photo lymts will occur In air, water, and soil. In aquatic systems, photolysis would be limited because of the attenuation of hoht with depth; in soils, photolysis would be rapid on the surface. A longer half-life is therefore expected as dapth increases. MOMS 219289 8-3 The persistence of these compounds in both water and soil may also be controlled by their uptake/metaboli sm by biota, but the degree to which this occurs cannot be de termined, based uoon existing metabolic degradation research. OeSTBUCTION/REMOVAl Since these compounds are both persistent in the environment and to.jic to animals and humans, more research needs to be directed toward the development of chemical, physical, and/or biological methods for effectively treating wastes containing PCDFs, PCODs, and PCBPs, as well as soil and water contaminated with these by-prod ucts. MONS 215290 Section 9 REFERENCES i/'iirtt * L .IT at. I'< rut iHirte irarct lMOU-tT*l*L ijit ano InvITOm- nfrL (HTtrtUTlOh Ct-14u? *-l*-a9 vtatMt t*t IMCt JCLt-. m , PESl'*r i*?4 I , a0 is tuiiO*Tt4n t.'tndTiOM ot tOLTCMt.4*imrit oittHloaiiaan* ti.pt. cucihi; tit* mCiptHT im caaaciro** confining ><h.vchl0*ih*rtt ntwftivu ih etn.'J'nmrri' otoxtn* hmd 'Clart* eontowo*. inaacr o rut Cv|>i)HHrr I) MUTIlN'.t* tr L CM llttnw atttl.CLNttotO N < I >j r 2l-iir tntat : .IT aL aOLVCHL'JtlNnTCP PtONIM* 'T'MSI, ftCNIOLIJ 'C0tS> HO ornf) OrtLTNUCLCM a*i>nari.,i i*c*H**> total* u*Ih act t|* cmEn ictiara ;i }*-i. r*M tapon i a ticul*tc cONraninoriON in me timsHaNTOn *t*tc grriu oiiilpinis *-*t* aiCiiMTco r ret sin mat. o*ll*i. rtxa*. (iicintct ii 'LCaniHi-, otftail On* aT th| *inch*ht4N crave Ottiet ttMLOmc iutochOCO Htu rota trait tteti ICLfatt tt* It I*01 01 wire.! J. tart ot *inCh*nti)n * TCatt* rOHCt (till UNCI*rata nf rutt Tint*, av l*.I*ti vounc.a t tut CMLOainaTfO OICtNlO-a Oloxin* in* tm* telenet 4* 1 t 9-r no a* toelarct aacnoxv Mttticioc*. u to*tv a* * t roUHC.fO* uilcv NfH TOOK.1**1 at 113-1*9 taraiL.O tOLVCNLOO INTtO t]*WCNVL*i OCCUtWNCt "# *10L0CIC*L ttttCT*. tttIDUC l(V ,44)l-ll,I17|. rj*aT*uft(.n IT at vijtHO, a aottOHIMC C*U*t* *v *IC( Oil CONTaninartO with aoLrCHtooiNarto *l*HtNVL* atma Hfatrn mo *4`is*l->t*i t*7i outf* n a ,c t**t ano a >.aaa aoLVCHLOOINarfO | liCHtonMaal '*csri> rouno in vumo OK ano In >i>Co j-aantti act cNiaot*Nt*t,7'4l**a* mi nja*TJUHf.n , r Nnwoa.aNO j na>sarana SOW* oa thc ttCEnr f|H*h>c> LjiKCtMitir, ruin* tm coNttotNct aaijrtcoinc*) Marionac coMttatNCt oh *uly,-HLO*iMnrto *law*HVLt iHovtntca nr*. CNicaco. Ill mo i*' taa-4**/*-?9-'H nnacn 14?* ta 14-Jt voiMinwaa.T .ano a irtoa saourn ot icnool Cmilo*IN vim aoLVCMLO* mart* IFNCNVL aOltONINC 0* VU'.HO (NvtIOM at*,, I? 410-419,1*?* (Hitcl.C H..IT at. eat cnaoaaToisaaanrc caaaaCTtaitTtct ot aiiTNtnric <.NLoalHaTt* IMHtemoaN*) lOtNTittcariOM Ot rva itont** m ant*lean -nt> laraatH *OLycmlo#|n*iio *]*wchvl* j a*ic.rooo cata , iiiiii-im, H?1 naeavaiM.v a auoaTtoNc.ano v aatuoa otTtantHaTiON ot 'm.otmaTto ortfiiJOtvaaM* IN IIMICIMII alio 'votHO OK' IIM.C (MViaOH vOHTaa rovh'ui Hi-U. t*70 notira.H ,tr m. otraKfO ftaniHartON ot aotvcMLOOINart* oi*(Hiotu*aa* in ac* aataataTlON* ano khncni vU*no oil *wu..(nv|*on conton ioxicol .i* 47-71,1*77 nrvara n ,r *a*Ntnor*.ao n riwira octictton ao 0(T(onih*tioh ot unti)0N D*eaorHLO*INaTCO CoaaouNO* IN aanCNI *IC( OIL C*OHO Thc 'VU4N0 * J too* NTC *0C jaaaN,l*'144>17l, 1*7* chin a N .it *l lCvcl* ano r.a* cttaonafocoaaaic aaTTtiN* oa aoLTCNLOtlnnreo *iaH(NVLt IN thc *iooo ot tariCNT* atre* act aoiiOHlne in ramaN OLL CNVIOON.CONTaN TON I COL 19, 119-11*.1*** HONS 215291 9-1 a T CHaac.aaO * 0 L'-r OL^CHI.atlHarco OlmfNVU ab F|)L VI"M(.0 r "'XCO 0ltfHoru*a IK rxe '3*K icE-**m jic raar Cmj.to *;* mijon- iC in rnicNUNC tuiL. Cu.if-'M LjNr*n roviCOt Jiivav-oj 'rC c Hafttotltirta Via tcri "CO WV acN aavafao co 'j"Ca UN|/ttt|. raCT uNf tVfOCN.IttZ jairMipt t fMf rtaontuN avo rat roxie Ctoup van uhivimitt ran* <i|U Ha.CM CONN I J? - v f ntaouCN a 0 tr at tr IPIli}tOCY *hO raraotOCY or a rirracatoaooi - ichjooioyih aoiiONiNC nch cavitoa Niai.rH.jj rr-M.itr? <(.tOrr|a * O CT at JCCVaafNCI or rcoo In tHvtaoaatNtat taartft raija ;,vTHH|jr altJdUll raHfnr|p at rNf ac* OIOXIN IvaaOOIUN raafat cirv nli>u*l >C*Tirol* i**J i.ararn v tfvitu or the tviar* iinich occjaaio in ttvijo in monih TduICOLOC(Cat N0 CHINK at airier* r CartaltNI a tavattatd am i nnlU. CO* trtertua *utLlcariOHt him roax <lt* rr i-r MNV a rin sHfNICNL tcrrat into"* tr J.i S-r aNO OIOXIN tCNUN ram Mix vaax I Ij aa itr-Jot Ulta n a rotrcatoaiNarfo oiatNjo.r-otoxia* aao o rtlajor'jran* ro*Nrida uLCutaciii'c aNO anairii jr imvlaoaaiNratir aaiaatouj Laaroiiaot ru o rNccii naivtttm or ijni* uata Iwimh nr* rata c n tr at. catoracai arria attt*t( or rcoo at ttvno. iratv in rHcooiaarco tioxlNt ant aciarco eorvaoUNO*, laaacr on rat inviionhihT j aoTZIatti.ir at .too rca>;.aoN ottos, Ctatroao.N v . nti. aa s*S-SS* acactta t fo c*lT(iia tott'trriCT altar lONiHiat i roo atcaaoratoaiHf acttirion.ata.jat or a uoariNC ooua or ixatrr* auttljato roa rat conai;-ro or tni ftiaoaiaa coanuHittt* or aoacanoa Mll(,t>ro*r Dor aa vat atari,t IT at nooiattrv aao ntra otricrt rtOM nr* to i r in rat rotULorioa Liviac in tni rcoo aoiLuTCO atta .'r Jtvfto IN' eatoataarco MOXiat aao atLartt coarooNOS ihtoi t on tni invinonninr o auTziNCtt at to* rttcarwa aattj.ttatroto. a v , uti aa sn-sr cr aarai.c aiaoar on tni ariatril* or oiNtNaaaroa tilCI SOOT Har not acriaaaN o c .IT at cirlMi.lM roa rat oitaooat or act* aao act ir|a* tv THiarwt onrtucriON (aa-4*a'j-t>-ajj ray.atoOMOO tiaca catir notuatv i **i 01 OorrCHIco ,i viviaao.afie c jaaaoal NfTNOOOtOOICat aaotLIN* in a<i*tINC J.J.r.t-TCOt (NVtOorMfNTat CONTalNariON ar tIVItO IN CNtOliaariO o i on in* aao attorn coaroum* inaacr on me invioonnInt o NurJiNoit.tr at tot attoanovt aattt.CtNtroao. h v .itoz. aa 4P>S4 Nlvara.N ,bno t ratNINOTO tNvttTtcarlna on oacaNO'CNtoaTNarto CONaoONOf roanco in aaHCNi oiet Olt TNat c*v*to rat -vu*no * j root nv* joc jaraa Jtil**.ttr* taro*lto.n a .T.a.ritamM aa r t. oarota oionIh* iaa-4*vj-at-nr. raoutroiat tavtaoHatarat atftaaca taoooaTOtr.CINclNNari.aovtaoto itat aotta.a a .aao e aarat roaNarioa or rotveatoataario oiotNioruaaat <acor*> raoN tni rraotvoio or iHOinouat tea ttoataa cataoaaNtat. t< i*r-i rv nrv aaart.c .it at, rootiariON or rotrCNtoriNaTto 0l0INZ0-r-tl0IN* <cOO*> no oiKNZOruaaNt lacortr or ouoninc oa atariNC CHtoaoratNart* Cninolaatat.r zT-jai.itro (trotito.a a ,tr at oioma* vot i totracit. taaoouat. raaNtaoar ino conraot tra4t*/z-0t-fSt INOutrtiat |NvltowNTat tttttocN taooraroar cinCINNOTI.JUHt toot. jamooN.o ,S tunoOTaoN.aao o aatlNO. *ornotion or rotrcHtoaiHarto oioinzov-Oioxih* oirtiac contuflioH or catotorataot roaautarioa* ;tt rorat tNVIOON ttlJt*-JIT, tro raowNar r. , aaa o Horziacta NicNMltrlc atrtCT* or rat ratanat roaaarioa or aatoctaarto otcaaic Coarouaot incluOInc rotrcatoaiaattt tlOCazo-a-oioxlN* raar n ratraocata icat otaiaaTJOa aao ottrrucTioa or Olttazoruaaat aao oiotaZo-r-oioxlNt roxicot lavltoa cata .5 tr-tJ.ttU 9-2 HONS 215292 Jl aHLlNC * ,XHO * LtNOtKOt INIIliqM Of CHI.OBINBTI# "CHlC t jtt r *><ct t rrOH '.'ON*o*TlflN INI CNLOriMartO OlOxtN* *NO Ml*rtO .)nF3UM0: |na<-r 311 rut (HWiaoNfltNr. 0 mutzimcc* (T i. . to* rcrcanON rate*.tLXirooo H y i*i m iti-ji* < cutterr c j rcaur caution* craxuu in rtatrtCTivt mown* uk*(iouvtNC thc |.;uti iotro uairtl aauacc .:<<!> 30-5? t 0i < LHHi'.ra h c r aaaOY a ortK* roaxarioN or ojocnzooioxhi: an* i}fHI* ;>]HCitNiart3H i)uulT FSuN NLOOtNartCi `MfMOL. MHO OfOJ.ari^ti IHYiaOM HtauTH rt*ia(., t<*('lN(NTau USUI 3 J-7 htj *1 |T*mii J * ,17 ai, iaWLIKC awe uaivtll aao'OCOL >iia atttOtlMC ortaaic taitiiOH* root! tTOTioNarr i.OOutriOH ioutcti lu txrotuat cvai'.>ar)OH nvt- *10" CONOUttlON CTUDIC3 taa-9tt/3(j.*ia,alOW*T ((((CM INtTITUTC, xautat City ao , itwitir not at aaaat e - a OUtt* auo a OotCMarer OlOXlNt. 0{OCNZOruOaNO. an* OTNta aoLVHac atto aaoaaricj aaooucrtoN utt. rotaariON. auo OttTavcrrOH Ma " a *0t lit i-i( 1r* ** a at .aa* C taaat 1 ctMt 1 a leaf r 0" or lUOtfiTUTtOH aaTTCtN* in i.HLOa IMTte OIOCHZO-F-010* I Nt tacooii tv aa*t irtctoonttov CHCNO- ..aert i i**-zi 1, 1*70 * outer a a aatraaarion or tuacirartvt irawoaot maturet or aoiTCNiortHarto ('liCHZo-r-MoittNf auc [-tttNZOruaaat tv ULTOavioitT ano cmm itraoiattoN or rut ocracMLOOO coaaoijNCt 4 cMonarou .nz<ltJ-Jt7 it7* t cootiT.O OKU MIlxnch aao a nOnc tuvltONNCHTat, ctHCtaTlON ao OtOtaoattON or OIOCNZOOlOxiNt auo tIttMZOrutaut CNVttON HfaitN rotate orttiMNtiH. 1*tut t J3T--0*.i*7j r cnovohoy.c o .r.ovrt.auo 0 hutzinoco. accwwriat in thc thom*. roNaariOM or CNLOOtNatt* C ONTOUNO I INCUJOINC aOLVCHLOttMTCt OIMNZO-a-O lOXINt IN 1 CNLOOIMTCO Of OX INI 4H0 Ml.lt Tto CONTOUNOt. I NTact ON rut (NVIOONHCNT o.NUTZrhcco.it at,,to* attcauOM root.tLNtroio,n t ,1*01. rr Z7S-J0I 4t unatraoN. a , * auotottON aa c aaaat roaaattON or eHtoaooiotHZOruoaNt Or roaaotatioN or -.m.orlaarto oiautNn. tnuit. CNCNOorucot 3 ii-ja 1 >70 Outtt N roauaTtON or MLV'NLOOINatto OittNZoruraHt 'acorn atto 01* MNlo-r-MOxiN* <rcoo*> raou thc ayaoi.vtl* or cinooottNZtHCt cnchointent t ois-ozo.1*7* 3t national atttaaCN council or caaaoa rotvcHLOOInattO oittNZO-'-iMomai cairttra roo tucta tratett oh nun auo Hit invioohncnt orrana i*ti 31 dllt.I .4 * a UltTtNNOUUt* an# 0 HUTZtNCtr roiYCHLOtINaTtO OlttHZO-roioxlNt auo ariarto coNrounrt in incinerator crnuCHr; in cniorinarco OlOXlNt auo tciarc* CuntOUNOt. laaact on THC CNVIoONNiut 0 nuTzihcco ft at .tot aerCaaoH rtttt.tiNtrtao, n y .itu. aa ;it-iti *z itottvi.a ,aaa o.aaacco rooaatiOH or roiYCHioaooiotNZOOioxiNt oho rOLTCHLOMOIMNlOrunaNt III uroau TNCIHCoaTOa CNttOIONt IN. chlOOIHartO OioatNt auo mlotio coNaowNtt. inaacr on thc cnvioohncht 0 hutzihcco CT ac .CM. aCOCaNOH a111 i.11Htr000. h t .1W1 aa jtl.jli 't outto.N a ,tr at. tOCNTir tear ION or aoLYCNLOainaTto orMNtoruaaN itoatr* in oly atu aao rca rvauLYtet CHtNOtrucoi.7 ait-rat.itrt. n ouNt.a r ,tt at. rtact CHCNitToitt or riati a tourct or cuLoriHarto OlOXINt. tCICIiCC. 1104 4400 >1 TOO-Jtt. OCT Z4.IM0. 33 artHUN t LtTTVt tryoT ON tTaTC-oaTuc-aoT or oionIh r*ON concoction tOONCCt aNtaiCON tOCICTY or nCCHaHicat CNCIHCCat.hcu Yoat.itOl 30 CaUHNCTT.0.0 CNVlaONHCNTaC CHLOatNNTCO OlOXlNt rON CONOOtTfON - THC Tract CNCNttTtltt or rire NvaoTHCtlt. Ini CHLOaiNartO ttOKINt auo ttcarte conaouHOt. laracr on thc cnvioonncnt 0 uuTZtHfta.tT at..cot atrcaaoN atett.CLNOroro n.v ,i*oz ra ztl-tol tr TitaioaN.T.o .CT at. caacacrealcation or toxic coaroNCHit in thc erruotuTt raou a otrooc-rioct ihcIniaarta rtootia cOMttav ,* >43-134,1 tat HOMS 215293 9-3 3* rotmtiNe o i rat ust or oioi" iooni* four oatioo to i(ltirt sonant no otrrwt oackcoouho livilo or oionini rn thc ihviroHHtHr, otvitu nro*rt Ne txTtNtiON or Tut aaijimt thcoov rn CHkOtlNATto moxini aho OtkATtO COHA017HOO. INAOCT oh TH( CNVIOONHIMT a HUTZIHCtt tT u to aiooanon Mitt.lirtroto. h r 1443 rr i*s-j74 It 0 ((*!.* V TOOCt CONTAN1NAHM IN *OLTO*OHINrtO OlAMtNTk not lITfag. *MT unO ;i**CN ro* runt i-i)H*OUMOO IN fNVIOOHNtNtAk >HH*Lti tULL IH- i*oh ONrM rontcOk .11 3. 1*7* 0* LUOTtHNOUNtO J U A I Oklt AMO 0 HUTZIHOtO ' H). O* I H* T tO 0 IMNZO-A-O l*( f~4 HO Otk*TfO COHAOUNOO IH (Hi. I Hit* TOO irfUUCNTt CNtMOAHftt, 4 tor-Ml 14*0 41 ! c .IT Ak AOkVCHkOOlHATtO OlOHIHt. OlHNIOrUOOHt *H0 OTHtO *OLTHtlC- LfA* OMTICt T0N10 111>: I Hf*A T I ON M0 AC* AlOft NIHmf frrTtNO to ootrooc' ' oittHTto otrort rut oivioioh or tNViooNntNTak CNtHitTOv ahIoIcah cMNicak IOC It tv rwtM CI Tv .HitOOu*! OtATtNM* '**1 41 HIIOIT.I C T COt TtO IA OOdUHtNT tt)0 ACOO tr*-44t/t-74-411. H*0t*CMU4trr| UOUOOH tOCtlTT.UNCOlN. JHkY 1474 f OOutt.C (T iointiaicotioh or CH.ootNrto ototNioruoANt rn hkicah aoctchlooiHATto oiAHtiiTus it*i its-j*r. 1 *73 44 r*AAt c ,ANO H 0 00010 CMtHKAk OOAtOMtt AH* ANAkYT IC *k "1411000 IK HA|.OCtH*TtO OlANtNTLt TIOAHtHTLO. NAANTH*LfHtt. 0 lOtNlOO 10* I MI *H0 01loTIO rooouert 0 0 KIHOOOUCn ID Clttvtto/HOOTH HOU.OHO 0 tONtO lr*L AOtlt.AHOTtOOAN,1*10 A* 41-7* 43 *4.000 a H H0 c t AAOAtO CONAAOIOON 0* THt CONAOOITtOH* 00 HOIKl.')* W41 an* AO0CL00 ttl* J CNOON*TOCO . 1*0>1*1-too. 107*. 40 N*TION*k tlICTI|C44 HANOAAi; TU*l*t AtOOCIATION *t*kT COnHCNT* ON MOT|r( 00 00*0*10 OUkCMNIHC CONktONINC AOkTCNLO* IHATtO OlAMfNVkt 'AC**) Ujt IN CLtCTOICAk tOUlANtNT HASHIHOTON, O.C . 10MC 1*01. 47 HOOITO.N ,4 NaoaCAH* *H0 C OaAAt AOkTCNkO*IHATtO OIOONIOAUOON 7l07 I rooHAMON roOM act ninth*! It hIaT an* ONY'-tN OUkk INVItON cm ran TO* I COL .C4'*#3*071,t*7* 04 kinoaai.* ,c *AA*t.*N* n a *u*t* ooonation or rokrcHkOOINarto oioiniorooaN* cAcooot an* rokrcw.0*IhaTio oiotNlo-r-oioitlNO trcootr r*on rut ATOOkTOI* or AOkVCNkO*INATtO OlAHtNVI. ITHtt* CHCHOt*Ht*t *'001 >301 >4*0 or OCHfCTt* a contamination or orrict tuikOtHc mitn rets ru**H* mOHN. *N0 OlAHfNVkfHft rrit AN tlICT*IC*k rOMOrOOMfO tUALOWJN in OIHOHANTOM. Ht TOO* TO tt ratOINTtO AT TNI JOO IHltONAtIONAL ITHAOiIUN on CNktaiNarto oionino an* otkarto coraowm*,:*4.i*uoe autraia ocroot* <0M 70 iNITN.O N .IT *k. AHOLVOIt TO* 1,1.7, *-TtTOaCH4.0*00 IOIH70TUOAU AHO 1 ] t 0-riTa*CM.o*o*lNii-A.oiONiN ih a toor oahali roon * roaNorooNlo I*loSION |H OtNOMaaTOM.NtV TOO* CNtHOSAHCat.Iti710-710.I!. 71 OAAAI.C. IHAACnttMTI* AOaH ACO-OOHHOt*. OK*n*|A-TIA* . I 7'4 > H 7 - < 3 I'47 7t OHAtM.H.H .AM* H.rtAH*. ANT0ICA4. AH* CHfHICAL AtOAtOTIt* OA OIONINO IH ILirlOH to THOit *I0*00*4 `0**rr> CHfHICAL kIhitico oivitioH.LlNTta roa CHON IC*<. AHvoico.nar lOMak ouatau or or ****. WHOM l ho TON, o c Tt HMCkOT.C k .C.L.atOaOl.AH* 4 ova* rOVTCNtOOINATt* OirnfAVl. truuot IN TIAMOrAOAft AAIHTCHAHCt OAtOATIOH*. an CH* HTC AOOOC.J ,43.171-174,1401 74 AkOOt.A O..AH* C t.AAANtt IINO0*4 *A*00*CN TO THt AAACTtOHATION AHO CkOOO Ot TOON I HAT IOH *A COHAktN HINTWIO 0* CHkOOlHOTt* AAOHAttC COHAOVHO* j.CMOMTKO ,147^30-1*4, 1**0. 73 chitTih.0 * .IT *4. CHkOOtaartO OlOINforuOANt AHO oiOCNZO-r-OIONino OtTCCTIQH AHO M*NTITaT|OH IN tLtertlCAi. tOUiantHT ANA THtia roonaTlOH OH*INC THt IHClHtOATION 0* Act* UtLkINCTON OCCINCt AOOOCIAltO OOCTHOOO. ONTAA 10 . OtOTtltOIO 1474 7* TATkOO 4 0.,AHO .N IkOTO CNkOtaCNt *00* J I .4 4 -TtTr*CHkO*0*10NTttAItHt AH* 3.1 ,4.4 '-TITOACHkOOdAlOOINZtHCt UAOATt AHO OUIIM IN CNkuOCHaTtO OlOHIHt AHO otkaTOO HAAOHNOt. INAaCT OH THt tNVtOOMHtHT 0 NUTZCNOta.tr tk .(0* AtocanOH Mill tkatrooo.H t ,itoi. r* sis-oai MOMS 215294 9-4 ** aortc c .a fttftOU.AftO * al-maiayi on tmc occaaoation or n 7 i-Ttr* '`.NLOAOQIMNZO-AAAAQiOYtN rr^OO IT "CAMS gA A MCW CL*S or CMLOSO* ionics CHvnott set tccnhol . u n MS lOIKM.r c ACMCD1AC nC1U>H *ho DISPOSAL MtctICM ro> VST(| iONTaIMIMO PIOMINS ANO KUlAMS TO 1C MCSCNTC0 AT TH( J*0 I HT(RNr ivHAosiuN on CNLOtrHArci oioxin* amo aCLArco conaounos,}AcziucrAusrti* OLtOICI 191* "9 pappi i tr n lOtMTiricamoh or polygmioaimatcd oioxini 'pcposl_ami MKNioruAAHS <acoa*> tm human ja*plcs> occupational cyposuic amo vusmo FintMTI AACSCNTCO AT 20 INTCAN#110**L UOAKSMpA ON CMLOAINATCO MpXjll* AMp ACtftTtO ''ONAOUMpS , AlL INR TOM v|tC|MlN OC I0ICA !*|l S A 4NAMMAMAM Cl |t ML CMLOAOLVJIS AALJCO TO TMt <?OMVCASIpN CMLOtOCAl- ION AfllOUCS trA.AAA/J-?|-l4A, HOCCMSrUNOC COArpAATfO,(MCL(MpOO ciiaai a j . july itn It CAOSlT.O S CIHtMUMQ INf NONSTCt rM( AM0TQCMCN|C*L OCSTAVCTT0M 0A CHLOA0OtO(NS 1M' OtSAOiAt AND DCCONTANlMAtfpN QA ACSTICC0CS H V AlNNCO/ CO ACS SYMA SC* 7J-I-H, IIP* 2 M(*N*or | OHOTOCHINICOL riKriwt 00 Mtnon do< >M 0I2>I*1 |m tNLO* IHUltt OufHOXV 02,101 N0 runt OlOKNt c tOHtL.CO (COL 1ULL tjrocutoun) it 21-01.1*21 1 *uinutt j * cr u. OH#roCH(Nitrov or 0l((N(0-r-0l0xtHl ih- cnlooooiiVMHt - ooicin tuo **ri m oloi*. to *o* cnm.m* ,i2i'*-*.i*2j *< uirr Hi , tr l ntLt toioli on oHotootaoMonoN or rcoo oh nan- T|OH trrtt lOtOVINO UtTN VUITMLI Oil. INi OIOXINi TOXICOLOIICOL MW cmnicol oookti r cottomni. chvih.lmo.oho c.colli.coo. io*ct*un ruOLKOTIOHt.HCW TOOK, 1*20. 00 (11-112. 4* oorot c . HCMLI.OHO r OL-HOIOVC TCO# lOCUOILIZOTION OHO ONOTOOICONrotinoH cn Mucout tOLurioHt cnvioou tci ticwhol..iiim-iw,<*20 00 WITH MCI#, 0 CT *L. OHOTOCHINICOL MCOOOOTIOH 00 01- OH* 0CTOCNL0M0IMN;.)OIJOM (NVIION HOOLTN OfltOCC ,(X*(UN(HT*L IttUC ll202-22l,t2I. *2 torn < COOO.ON* N omit IMHT101 cot I oh 00 OOLVCHLOOINOTIO OIMN202UOM* (OCOOlt IN COHHCOCIOL CNLOOOTNtHOL 200HUL0TI0N1 CHOHOOONOM, 2'4*l-**t.1*20. *0 "tout'd** 2 .OHO H J.OWIIT truOKO OH rut OIOOCCVHULOTION OHO HtCOOOI*L OCCOOMTION 00 l.I 7,l-T(r*OCUL0Raal((H20-O-0l0XtN (NVIOOH NCOLtN OCOIOCC .(XTMIMNTOL I HOC *i 291-200, 1*21 *2 TOUNC L ,C ( TMOLKCN.ONO V 2 CotOHCV NttOICIOI OtMCt *IT( TO(fn(NT *H0 (NVIOOHNCNTOL HOHITOOIHG rO-2*-l*.U * Olt 200C1 OCCUOOTIONOL uho chvkonkntol hcoltm LMMtraov, itooxs oro, rcxii, t2* >i voro c t .mo r mrouHuoo tort or 2. i,2,*-T(TtocHLO*oot*(N20-*-<>ioriH <TCOO> IN O HCOCL OOUOTIC (NVIOOHHCNf. OOCN (NVIOOH CONTOlt TOXICOL .2 ?4*-H2, 1*2* *< HflLINC.C 1 OtfTICIOt HOOILITV IN MIL II OOOLICOTIONl 02 *OIL THIN Lm T(t CNOOHOTOMOOHV. MIL 1CI IOC OHO* OOOC ,llt2J2-241.I*2t >2 NOLLIHO.C 1..0H0 O.C.TUOHCt. 0((T|CIO( HOOILITVtOCTCIHINOTION IT SOIL TNIN-LOVn CNHOH02000OONV KIIKI. ' 02 1102-101, I *00 II NONOtOCIO.t (T OL. TNC ICVCIO OCCIOCNTitTl NOTUM, (XTCNT , (HO C0N1C0UCN- C(1 i occur NVO,,22i222-221.1*21, ** klOONCV O C..t R, WOOL *0N, OHO C O (LLINOTON 0(*1|1T(HC( OHO HCToOOLIlN 02 r.HLOOOOtOMINl IN MIL*. (NVIOOH, SCI TICNNOL .0 101 2-101. 1421 *1 CAHONI.I ,(T OL. LOOOROTOOV IHVCOTIOortOH 20* TNO NICtOOIOLOCtCOL 0(0(0- OoriON 02 2 J.2,t-T(rtOCHLOOOOIOCNZa-0-OiaXIN IN lOIL OV OOOITION 02 OOCMlC C0NO04T INI CNLOtIHOKO OIOHINI OHO *(L*T(0 CONOOUHOl. IHOOCT ON IN* (HVIIOHHONT 0 NUT( INOCt. (T OL .(00. 0(M0H0N 00(1*, (LH*20*0 . N V I'02 Ot 1-I|I, * k^ONtV.O C..(T *L. TtrtOCNLOOOOIOlNtOOteXIN IN TNt (HVIRONMNT i lOVKCt. 2*T( OHO OICONTOHINOTION (NVItOH. HtOLIN 0(*(0(C .(XOMIMHTOL HOW 1 22J-227,1*21 HONS 215295 9-5 ir >> * 1 0A * 01 In' i 41 i*4 I A* 1*4 >AF l*D 1*4 II* III tiz Ml ii* ID M<jtrrt* r and " fhilIffi lnoofaiof) condition* to mfit uii tur (NviroHnfnt f.jtc T ,')i< ff dz-u iruoict oh nicxoiial "(tadoliih or t.-dp '.'ncii ML'jniirrto oioxih; aho fclftcp .ohfouhPi o Nuct'ii.ct ct al cdi fcdcahiih f*c;j flh4- - i'ihi, l w j CAiAHfr and chfhhCll rtrttirtxct hovchcht and ptlutirtit >Tuftu or repo mi tofa( jitcj hftsivclt coNfAHiNArcp huh iheiiOii HfiiiclDfl 10 It `('CHtfO rr me jfp i n re Iha 11 onal Tyhfoiiua on H. II Iiia TCCi O 10* INF AND *fL*tCD CONFOUND!. TALZDUDC AUTTIIa OCTDFCD i *AJ roiotr m ct al iraucTutf Cl"CIpat:oh or hahhaliah tcoo-hctaDolitci t'l-iOHO At ,0 iNTfiNA | IgllNL XOIklHOF OH CHLODMlATCO OIOXIHD AND >(L-tCO CJHFOUHOD AFlJnCTOM vIDCIHIa OCTPDCF i )( MfAMAH T I) :hl0I00 ItfNFOD tOXIN* AND CHLOAOOIDCHZOFUDAND ah Overview orin orrotr:-ArioN nazafoouo ha>tcd j cxhcd.co ann addof tCICHCt ANII AAADA HICHl'iAH 'HZ FA Z4Z-ZD* 01 OOHFMira A C VtVfANn AND C IFFFOnI CHvtAONHCNfAL rCACtjTFHCf or i J - x-n-pp At ievCFO III NLOrtHAtfO OIOXIHD AIIO FfLA rfO CONFOUND! IHFalT .<1 rwr fiiviroHHCHT o HijTimgcr ft al foz fcdcfhon rreti clhxfofp n / I -W rr I *i-i1 j FltHHfA J A AHOTOLVIIi Or ''po HHO TDTFLUFAL tH OH XLItA ahD SulL FULL I'li-IIO* f.NTAH TOXICOL <4 *F->Z ! *FD -riCDCL o mr OtOxlnDi toxic and -Itill r*0<J(LC*onC (hviiohncmf jj -IJ I*01 itfHL r u cr At THf jraAILIty or *CHrAChlooofhchOl a0 chl OF InntFO PtOtIMS Tl) JUHLICHT. HCAT, and OINO'JDTTOH IN CHLOtOOIOXIH! - OOICJH NHt FAt( ( H (LAID.CD NOV |'H|H >( I ZD II *-1 ZD. I FT JtftlOtF.il t IIOACCUItULATlOH 1> J 1 t (~ tf rFACHLOFODTffNZO-FAFA-O I Ox I ii |H INLOFIHATCO FHfHOXV HLtPI AHD THf| OIOXIND C FAHCL.CD (COL DULL 1 ItOCXNuLH),*.' aDS-ZDZ lit* xCTTt JO (1 AL. THC COHFOfltTOtl OF FCOF AND FCDO FTOTPUTJ 1H JCOITNt; t TH( NUOtvH AHD HOUlAtiVMli Ft,(At FAftCHTCO At tM( ALA DIOXIN JYHF04IIJ" tnxiAi 'iii hi Ioduai .oerriHDtr 1*4* A FCC | AH I 0 rgxyeOLOCT i)F fCD AHD FfLATCO ""FOUHDfr OD|CvATTOt IN HHH IN cHLOAIHaTCD bluxIHt AND FCLATCD CQHF -JUNO t, I "Fact OH TNt CHXIFOmhCHT 0 wrtiai'ifl Cf AL et'J FCFCAHON rlflf .(LHtrotD H T III! FF 4*1.401 Aljrr J c IT AC LONO-tfFH HAZAFDO of FOLOCHLODIHATCD DIDCHZODI OX 1 HI AHD FOLfCHLODIHATl# 01DCHZOFUAAH* (HVIIOH HtALTH F(*DF(C lD'2Zr-Z4*. 100* na TTHIaDCHX 0 IUOXCV AOOIJT TOx ICOLOCICAL DATA OF Z 1,7 0-T|TFAlNLOFOx I * OfUZO-F-DIOXTH t TCOD ) IH OlOXtN, rnJICOl.OOICAL ANO CNCHKaL a tr f C 11 r 'ATTADCHI a CAVALL**O.AhO 0 CALLJ. (01 ZFCCTrUH FUOLICATIOHt N(N torx I*.-* rr izj-m ni'COIIHCLL, ( ( .AND J A HOOF! Tn( TgxICOFATNOLOCY OF TCDO |M DIOXIN TAvICOLMICAL aHD CNCNICAL aO*(CT( F CaTTAKhI a CavaLLnFO AND 0 NALL I CFCCTDUH FUn.TCDTIOHD.NCN XOIF lOFD. FF I]F*I4I HtNL.t A ,F U.DCATrv.AHD T a CAOICNICZ DTUDICf OF THC TOXICITY or Z.l r.A.rcTDMIHLOOWiaCNZO'F-oioxiN (TCDD) ahh n y ACID DCI .JJDiZ*4JtJ,tOTO JAH HILLCt.J F .J J LFL1CH (MO i ALLfH IHCDCDOCO IHCtOCNCC OF N(0Fi.aIH* IN DITO CXFOtCD TO LON LCVCLf OF Z, J , 7 , C-ICTMCHLODOO I DCNZO-FOlOXtll CNCH0DFMnC.4-lIF.DAA. I HT AAFFC.C .CT AL AHHLTDtl or FOLYCMLOI IHATCi 01 DiH Z0-* *01 OX I HD AHO 01* ICHZOrulFHD FlCDCHTCD FT IFCCINCN DANFIHC UODXDHIF DFADDOUCXCH.mCDT OCntFNY HAY lODl, lAiNT-auF.c tmc dttucrote aho diochchical cffcctd or rcoo in DIOXIN- TOXICOLOCICOL AH* CHCHICAL ADFCCfi F CDITA0(H| A CAXALLAFO AHO 0 CM. LI. COD DFCCTDIf* FUDLICAT IOH . N(U TODr.lDF* FF lF>l*l HCFL.D .AHO F (CATTY IHFLUCNCC OF CNZYHC IHOUCTlON ON TCOO TOXICITY IH 01OX INi TOXICOLOCICAL AHO CHCNICDL ADFCCT1 F CFTT#|H| CNvAlLFOO, AHO C CFI.Lt,COD JFCCTFIJH F'JOliCaTIOND.HCN YOal.lDFO ff 141-111 9-6 WINS 215296 M7TC.J ,amo C otooauc cu*Ncri :h aar xunc elLL nrna.M,j 0u*.u<- i i, ?,-riT*cHLfl*o-tti*iH2o-'HOi(ii* intoxication in. tiimy rojiij.'a,, ChL xfl CNtMICtL nSatCT4 t CATTattNI n CivALLaO`7 AMO C C*tLI ff.* L0,l` jrtCTKin ufttc*riuMt,N(w ts*t `79 rr iimii c'OlCta and ICNLAlTfa -It 71y-7h*.1*7* tIOLOGIcac OIGOAOailOM or rLoo in raij ,,,ru.| CattMCCt X r AND ffO noLtCUL* *(! >0* Tu| INDUCTION 07 "AYC HYOIOCArtOtl MYptOYYLAit HI OlOXlH T u - I C OL JO I L *L UNO tutnicac -inert r Catt-*eh| CavALLAto AMD 0 iiAt.Lt irtCTAUN aijti.ua* TlOnt Ntu Yijar i*7* a* iij-i.i me nr an autrur* and I a tarn rare moctnrti I HOatiAY of cnioaiNarrr CIttMloo 10*in* ap *tL T(t Cmcmi.-al* tHvitOM hIactn attirtC InattlnrnT*t Itiut * 1*1-17* n;i riLA'in a .amo ( GLO'fa nustlt in >( ntcn*nt* or toxicity or rot tutorlMario otatnzo-a-cunus cnviaoH mialtm ataiatc , txatatHtMTAt IttUC 1 149-191,1*71 naora t j . j iunmaav '-nHr*a(HC( OH 0lOtnroo 10*In* Am DlttNioruOAN* nationAt IHtTtTUTt or IMYItuHMfnTac n(at*n i(awiC({ nAIIL a-1 i 71 (mvioom miactn rtttrtc tiattminrAL ittut s ir*-iti,i*ri HiuoitT o ,(T ac. a tuaviv or tut tNotvotoxtc trrtcr* or rcoo m mammali- ah trrcitt tHvtoftH ntAL'n atatrtc fxrtotHtNTAc lltut.l *7-7*.t*7i moat i i .IT At aotmarac Itricn or m*T(*mac Ktotult K I 1 1 t-rit aac mc oooo t Mnio- -o 10 x hi 'Tcoo< Inviiom Mtacn* atatrtc txatrintNtaL USUI ? l-as. 1*71. 70LAM0 A .AN* t CCOVt* CMCOIIHAttO *IAMtNYC INOUCTION Or AYL HVOAOCAA|4M NVOOOXYLAtt ACTIVtt" a iU'OT Or tut tTtUCTU*t-AcTIVITY ttCAttOMlMIA HOC ANAAMACOL .11!f*4-*JI,I*77 KNwttt * * t* ac toxicolocy or enco*iM*rt oiatMto-r-oioxiN* in CHCoaooioniM* o*ICIM AM* rart t n *lai to **v chin t* Wli 99-44 1471 tciwtTi a a ,tr ac. toxicology or CMco*iiiATt* oittnio-r-otoxtM* tnntrOM health notate , tvaiatHtMtAc Ittut 9i*7-4,i4Tl ACL IN. J * .IT AC. HOtTMOCOCICAL CMANCtt IN HOMrr/t CONtUHIMC A OUT COM- tAINIlie CON Ltvtcl or 1 1 7 -TtTOACNCOOOtltfNlO-t-OtOXtH root COIMtT TOXICOL .I* a*l-*lt,1*77 MfMUCTY N a ,| N XOHfOANtl,AMO T J rAAttLC CM*ONIC TOXICItV Of 117 t-rtTOACNLOOOeitfMZorutAM r,-,* annul HACAOUti IN CNCOOTMarto l-IOuiNi ano ltc*7tt tOnaouNM ImAa. t on mt ImvIIOIihCnT 0 nu Timet* at tot aticaiMM r*t`j> tLHtroro H / 1 >4i *a 411-air t* Hfnat J A .IT AC COItaaaAtrwt tomiciTy or rurtt nACOCtnalto t> I tin 10roanMt IH CutMCA net n|Ct, aHO (HftUt nontfli ana u T atnt it t 111 191-1*1.1*7* C*ON I 0 CUT am*out trrtCTl or cmcywacniictHt IN cnlo*1matto OI0XIM4 AMD atLATtO cowaowiio*. IM4ACT OH ThI INV | OOMHt NT 0 WiTttnr.f* ft At . tpl rtacAHOM a*tt*,tcittro**.H y ,14*1 aa 919-9I1. MOOAt.l A. TOXICITY or 1.1.7 t-- TfTAACMC0*001OtNIO-AAAA--01OYIN IN CHLO- * I HA TIO AM*MONY AC tot Alio THI I* 010X1N4 C *atl.lO tCOL l)LC '!TOC- NOCHl.ir1114-144.1*71 aocCNlAAT.r , v * itamo. amo * lanartar hunah HtAcru trrtcr 1 ra.%n acciornrac atLtAtt 0* HTMCNCOtOOIttMlo-a-tfOXIM <TCOO) AT ttutto. ITALY amm m y atno *cI .lllilli-lit 147* OtCAO c.n ,L t OIOMOAUM.AMO H * HATTHtNt tltaoOITTOH r 1 1.7 t-T|T*A- CMLoaoorotMioruAAN in cutNt* riot, tart an* HONxtY* in, cncoa1marto OIOXIH* AM* Itc ATI* CONruljHOt, IHAAC T ON THt IHV I tOMHtH T 0 HUTIINCtl tT *C..tot artcanon aattt.tlHtroAo.N y i*tt. aa itr-iit HAtUOA Y AN* N KUOOKI *i)LYCNCO*INATtO 0TOtNlOTUOAHt AN* *IC*r(7 C0HaoUNOl IN AATItNTt |TN -VU*H*` IN. CMCOOINATtO DIOXIN* AN* ItLaTfD 'ONaOUNDt. IMAaCT ON THt tMvrtOMHfNT o MUTlIHCt* IT AC ,101 attCAAON aattl (CNtrOtO.N Y 1*03 ft *41-9*4 HONS 215297 ! 15 ii 07 in >* in iJ, nr 14a 14a 14a 1*1 i40 hi tot in >12 51 aarrf C , amo a atcata 'nmm, aaawtin or auraa taaFLit. :OtarirteaMow uMO OUaHT | F TCatION OF BOL rCNLut I NalfO bJOXIMf OHO (l I IfalOruaaal B4fl(MTfO ar TH( u I -iwfOltN wornwor aat HO] caott.a L ,ir at n(rui}o tLt'l(n roa rat OfrtaarHarion or TfTaacMLorotilOfNiaoioxla ar rut low raan rr raiwilON Lfvfw Nat CNfa *j m/. HM.IIII bonlbmo a t .aa c c taac ratrarariOH awo caaaacrtatiartON or cwwoai- a t(0 MttNIO-F-OIOXINi J (ttC F00* CNtH , ft i I IU-1 144. t 11^ WlWLIaaf.O T awe J ILauraFIfwO tCFftaraii aftHQO BOO raf OfTfCTrON Or LHLoaooiaiNio-r-oioxiNii m rH( rtitNct or aw oaooi rotate t. CNwotowaraTaawtNft, aaa CNWOaof IICHZOrijaaHi j attOC orr auaw CNfN ,51-01- It. 1070 ririiroHt.o it at Ofrfaariiarion or roLVCNwoaOMOtNto>r>eioxlNi ant atLrto cobfouhoi in cOHnftclaL t, wLaaoaafaowi J aHOC orr aMat CNtH II 41-11.1411 WOOL10a.I a a r THOaa* r 0 J fNtta tuavft or rowv(WWOaoOIO(H20-r. 0IOX1H CONTfNT IN ttlflTfl' FEirtCIOtt j acaic rooa cm a , 141J51 - ria ma MUriiNcca.o f tart awo v nrro nwawrtit or cMLoaiaarf* aaoNaric ateao. caatONf fv txNWftTivf caLOaiaariOH ouawtrarlvt awo ftaucTijaaw aiatcrt of rat rtacNLOao-Dfaiwarivft of tlFafNTL aeraraawtNt, rtarwfavw. bllfato- rutbn, oiotatooioxia. ane oof iwr j (NvraoN aaac cnin iitt-iia, nn aircHUN a x ,N a roariiai.wf r ao C r 0Ltr araotFHfric rafttuar loalzanow waft tF(CTaON(Tr< raf I loaf irtciric Of TfaNfHar ion or Tract LfvfLt or i I F.a-rcoo in THt ratitHcf or conrawinonto rafttarte ar lira aaauaL HttTiwc,aatIlbn toctfTt for wait incTaoNtrav,aiaafaFOL l rm tauewaaa a ,awo a a(tfw>4a aa taaaovfo aaaw tf is roa rttaaCNwOtooilfUZO' F'OIOXtai ini (NWOOeOluxiNt . oaiCtN awo rare f a tiatr to aov <.mCh ,ra Ut.11-i t. till rftra.L n ,fr ac a Hfa ofTfCroa roa tcoffaiwc reto awo rear laartfl aa(-.r>iTro ar Tat ioTa Hariowaw NffTiae or rat aacaicaN CNlNtcac tocifir aaaiat e r it, n r ttour r, lf rtaota Hal cauNNcrr w o ,awe a n tttww otrtaaiNTioN or cwwoaiNarto oiatHio-r-aioKiNi awo oiatNioauaaat ; vaaiout aaTtaiawt tawiaoa atawra rtrtrtc tx* tftlNtHTBW tIUlt Si 15-11.1111. totta.H a awawrtlt or rowtcwwoaiaarto oiHNlo-r-oioxiai aao oitfaioruaeat la cNwoaiNOTto lafaOLt It watt raacwtaTOcawaat j iHBoaarata , 1 115-llt. Ills ttatta.w a ,tr at. ofTtaatNarioN or caLoaiNarto oiatNio-r-oioxiHt ih ttaracwwoaorwfjiow ot oat karowaTocaarNt watt tateraoHtTat aaaw cata ,44-144 103,1411. aiMati,0 N..1T at. aaawtricaw afTHOft roa NflOldOf orawot /OL I OfrfaNiaarioa or cafaicaw coaronrioN atroar no ra 15-tiit.u 1 aia roact trortao conmm.aNOMNi an. vaowioctow. o c ,aat ms taa.t.o .n L ran.oo.awe r o ritaaaN ctrttaiaariow or 1,1 i.t-rtrracHLOFo0iaCNl0-r-0i0aiH ottioutt on hctol tuaraett It iC-at CHtNotrNttt.t 1-14.1414 lanoa.N l.,(t aw. Lfvtwt or TtToacNWoao-oiotNio-r-oioxiN in taviaoaNONTaw aao oiowooicaw tawrttt at oftioninoo It cat caaoNaTOcaaONt-aioN aftowuT 10a watt tricrioatrat rattlNifa ar lira awauaw aftTiac amaica toe iftv roa mt trtciaowc tov.ioh oitco.mt. cawawwaoo.a.,tr aw. taNOLiwo, occwaatwcc, awo tvawoaTioH or rcooi awo rcort raoa laciNtaaTto town uataw watte. CNtNOOrNfot.titn-tl*. i*1 IHaoorr w a ,awo a a Hijwwtw. TNfl OfTfaalaaTrow or l.i i |-TfTrtcawotoolOfNio-a-oioxtN in ttowocicaw txrracrt at eat cwooNarocaarNt watt iBiCTaowf rat otoaato watt trie raoa., iii-n.itit aaawiat.a.L ,fT aw. tawawt ratraaarlON awo cat CKaowarocaarar-aatf trccTaomrat ofTtaNtwarioh or i.i.i. o-TctaacaweaeoiitNio-r-oioxiN aaaw CNfa .siimi-nil,not HONS 21S28 9-8 '34 lOten.j c ,IT M. ee-a* noeitwi, *0. .................. IM1 tWM IN p**ncuc*n* 7*0, MW*t 4l(Tt4fi. IBI4ICN tOCHTv * *t CHLotM '*!at > H ouiio * p#cvcwco*it**no i(io--oia)<mi ritanow 0* 1*D*I*< *v ta* CWOaarOcaaawv-..*..iJT7*'.** TOCO . 1*79 ** '3-. U* t. .PM* N P *0Hapor PULTCHLOdCftt IN1*IO-*-o,o, ... rlr'JPawt UN* OIMZOCI 1M CP afPI (OftftUtMlC* 1HBU*TIILLi' 0,,*w- *(HVt*C*aMCaCt HT Ul IttuHTH* wv .*I,|**|.,*,,*** >4r OuOia.M ,**# c Oaan <*i4P-tocunoit tal i-NPoaarMaaawv or tm u 7ltaCML****t*tHZ0-a-Ol0X|W 110*141 awac CHIN .11 1117-1141, i Mt *" "* xapat.e .CT ai 1KH11'IC411W 04 POCUNl 0*1 waif* (CNIorMWN tPCOPtl *(T*1iO In OalltMT* VtTN vutHO CM*N0*PW**. PiZO*-lM. 1*7* <11 rijPtat.N aw* T aaaU** 0(T'(aI*aTI0* OP M.veH4.MtMt(0 OIMNfprtiaaw I40MP* Mt*lw0 It* Pal Han alTM vu*MO CWOWOIPNfat. T< 771-777, nr* 144 rtrtHPM r o ,1' , iwctHtaanow 04 ewoatcac vaont coaTaiame polvcmlotlttPTtO *1HtWVl*i ***t***(l Of TtfTf COMMlCt** *1 NUlPl (MVIOOM"tmac ifavKtf. ftna pat* rcxa* aw* tNIPSc *r*T(M cONPawv. tc toaaoa. prnapfal IN. OtTOXt*(Cation Of watMOOUO **1t j tXNCa.fo aww a**** 4C ItKI.aNM aOOOP MCPItM, (Ml. PP 141-1*4 '4t i.aapaai(l ,c.c. .aw* r t alOTOte*. otttOPiwaTtoii * Ttvaa- hi**-, ac*raaw atTCiCC*OOI*t*lt-* -Cl 071a Itoata* 1m P*artCUL*Tt *an*C(0 ar Pawn PIO TaiLLIVN CIVIC*. awai-.CMtN .111 >41 wM,4.t..lT ac. o*Tt*iailow or POCYCNcMiwaito oipcNZO-P'OIOpih* in tioctctcac taa*Lli *r Miami iwcaieac lOMitauM mii *pt*owit*y. *w*C .CHC* .**t I47*ia7*. t*M 41 SHITM.LH .IT ac. IMNHPICartON aw* *u*MTtT*riQH of PQCTCHCOPOOIOIKZOpiiaaw* in tMvtaoMMNiat. iaNPiti poCMhTCO at l*T awmac wftitac.aafPi- cam miCTv ro* m* *PtCT*oaT*Y. miw**n.)f*i. 44 **#P< c .IT ac. iMHTtPICatlM OP aOCTCWCO*(Watt* olMWIOPltM** tN (W*MMMMWT*L **N*CI* H*H|Ot.l*I< IIMM.IMI >1 DMtWM.p IMiMt a*MOTION iHCTIt OP aapaaTlt ITwta CONPOtwO* tt CHMWCT(*I*TIC MM* op 01*I*10-P-*l*tH M*tv*llyn. TMUCKU 2*0*4(1.77' 'MIS. IW mi awlctM.o aaiaaaauoa op C*(.o*0io(itZO-4'OlOxiw* apt Tpwicococteai. IVaCUPTlON IM< CMCMMIOXIW* - 0*1C1N 4M **Tt C.MOCPlX.IO P*V CHM * .11t.IM-lll.HTl '47 Mix.*.* ,ti ac. (PWPMlsi* *p tpicipk Pocrcwcoatwario si*aio-''OIANIW* 1.4M.CWCM. ,4l. 141*.J4J7. IIP* <44 w**itca.r. J. ,c,c caaaa*tt.awo a it itihc stntm(*I* aw* totwrif ic*tioh P TMt 11 TtT*aCMC**I*<N10-P~t'iaxiM ttont** *v wiett HP*o#aaact Clio:* CMtfMTOMaawv an* M* ctw*i**** aaac twctt .5' iitj-i;i up* 144 L*apaa*pi.c.c..aM* t,j.wcTtc*. *rrwcti* *m imntipicptiow *p m ' ntwKHC9M*(CNM>*'*l9xin lioai** *v wiN p*POaaaci ctMi* aw* ccWWW * cw**M*r*ca*PNY. cmw>pw*i. t *. i-1*. i * i. if* UMPMMt.c.C. .an* t 4.wctTPld. P1CM cacoaiaaTjea ppoctouat poa twnn op wimm* CMCMitMTt* *[*m(o-p-oi*niw* pow [ucw.j.p.a-nT*aeMLOM()lMitIO>*>*(aatM awwc cm*..*4.4*1-44*. 14*1 '71 CUH*...IT ac, #TPTW*1 of cc*lwaTC* *inwlOPuaaN* * pwotoLT*tt *p ewcoataatt* oipwinv* itnc** in actrowi **cvttut. c*wo*p**i.t. n?-m,i*7T <71 ***(.a a ,a -# **** r.aw* c aap*c Poaaatloa P POCTCWcoaiHaU* #iMPiPu*aat* iPtr> Poa rw* pvmct*!* op act*, cwcnoopnopi.p 1*7*. <7i oaaa,a..it ac rmTitcfi* or ttaceeiwoTco oipwcnw ttwot ** oioinio*U**W* > a 01*CV**IM OP IPIctPtC tiowto* avoicaoct. CMCmtPMCOC. <it 1*11*0,1*01 MOWS 9-9 in > ?t <' 't 'a r i#a Id I itl i at i ga It i a* nr taa tat iff it< 111 fl it aaai ( .aag u * totta aokymkOatnaTte otomna aag gtttaiorueant In tMktnfanfOt tatkUlNTt nntta r (il cnaancrrattarton )f rtTaneakoeogratMioaunnM ! mr [yrtMoto trailer cifNrcc ttagat rut givitign oa iHviaonninrak yntnrsray.antaican catmcak lor.ttry n*i*r ctry matsuai Jtartntta itat frit)Tout o otattatnartoa or aokvCnioaootatnlo-'-eioytnt ang agkyCHLatnotafH*0'*ua*Nt tM idnnfactak dlkarrnt #y cat Lltuto ettenaroeaannv j mate fooo catn it uM-ijaa >trr nine.* j i a aogrt a tallt ntra rot attariac nt nfraggttt" ga afragaraefoagain ar mcaofdnat luactkkoa* anacrioat ami aieCNfH tar' ltdt kaataatcl k k ,f t mantc* ne a a trtnk etriaMnarton or iMt-itiraikkioa conctnraartons da l 3,r a*rt TaacnkiMogiatnzo-a-o jiia in ana amt. (Hta Si lancagatr a i aim i tnagoat atriamnarton or anari-ata-ratki.Ida cgncturaartoat da reran- at**- Htara- ana gcTncHkoaooioafMzo-a-orairi la ngana milk an*k entn 1: zm-ataa, i*M rttanan r g ,tr at oiaaiHt vol i analytical, ntragg ago mount rL uattat faa-att'l-gt-n?. maul ra rak (HwiaeNiKMtaL atltaacn Laggaargay cmcmnnM, jut naa vaMKft.fi a et at ItlaaiMkOtOOratalo-a-0lorrat |M cnlmcnk uatrti ogtous taatutari ana toir.l Laenoiantat.t'iii-itJ. itaa tawoa a k ,|( ai atilt<ntnif oa mciaeaanon aagcttltl ai tggaert or jgatargne 'ntoamatag nvoaocnagant cgactHraaTtgai or Ctet'itai mo ioitllkf arcuatoa eenaegnot in rnclneanron eaaigfnrt aatttartg at rat .it Iona ttaaetl un,aaagat ulty hi itouai, ttartnata itaz agtta a a , a n aottHaagi ana c aaaat return (canon ga ngkrcnkOtmarre atatutg-a-giaain iton*at aouno in akr ala cntnoaaafat. h itt-iti uri aaaat c gniwtattrr ea unfa aeiouai* aga knit ga acoat ang acort IN aiv atH atatonak connontcarioa novrngt* Hid naTtOMak atttaaCH cddncl> or manga agkvcnk-ja(Hattg eitfNdg-n>glearnt Llmrationi to egaatnr anmvrKak rtcaniagt* otiana itar ntatcan catmcak toeittr cgtotklNtt aga gara aeguitirron ang gara ggaktrv (vm nation in taviagantHtak cntuttray anac cat* it ttn-ittf tat. aaunkfy.w C tt ak. kOd-attOLUfron nguriakt roa otttettoa cat cnagnargcaaanic-natf latcragnt rate eonanaijon da in (tramrlOa-Lktinga nirnoot aga gCTtaniniHC i.j.r.a-rcraCako*oeiatneo-a-biox|H m >t. j ^.a 1 c agog chin ,jt-taaa.taaa, nai tggct.a a a ,fT ac. ttraacrtga taarcttacltt or aokrenkdtinorio grttatea-gioaiat a^ aakrcakoataartg oIatato-auaaat aaga fLr n>a anaL -.nta tltatiaal,Itai taaa.a. tttiattt ga agtvcnkgainaTtg gietatgagaaat aa g gnttarangn MivtaaiTT oa gaca.tgtata, nat aaaat,e.,t naatkoaa.aag a atacevltr taaaataatrta *aaa nagaa act* etatNOtt attfOMM. coaaoatcarron itaa. tMrrft.r ,tT ak. aigaaakvtl* ga nukvcnkaataartg oiatatgauann nag giatntneioxtn atxTWtt in aLf ata aatttarcg ar rat lag laytaanrigaak tvnaggga ga caLoataartg oio*mt ang atkarta egnaovagt taLtfgac auttara ocrgaca i >aa mg t ton, j c ,aag a agkaag ataarmitariga ga aggtt rtaatgna ctkk u[ f a a00UC (0 ar 1 ],r.a-TtraacnkgaoeiatatO-a-Otgy|a aa in mao nootk oa rgxtcirv ctLLitt.ta-ia i*at gattw.tt a a .aag a not aw aucktaa uaraat oa t. J, r. f-TtraacakgagorgcnzO' a-oio*[M in csrai./ai aag gga/u met, aokt ga rat ataarlc cvrotok atccaroa aaoTtta j giOk.Cnta..tia,taia-tati, ttrt taaakaw j .aag t k catrtaktnt laggcrign ga tatvnt aeriviry in ctLk CUktgati a ana re tetna aoa otrcciiga ga manat aokvcNLOaiMaTto OtcaalC egnaggagt j attoe.orr aaak entn .tt, ttt-rit.mt 9-10 HONS 215300 IM MNUTtOH J [M VIVO MO I* ho ccctoctcfti VtTftO I** ifjcct* n ) i t ** tiACML0iOitH20'-mo-mh TOffCOUJft* Of HHLOCKHftTCO WVP#',4**0 * >) rnpu mmo *tontow tot rO*!-'i ?v ticv vnir.i*at> pp ffr.: '*4 J ' ,,W * !/rif 6ITCC_TrI0rtHVin**M<*0Mt0VfftfHO-Lftlft0* **CtTf|vti^tCv U.jtmHifl6U t*O>F mH e|tVo|Tf Mi#o*t*w!!-'I tTM-t ; "< *UIL(He Hi* ir.ft HONS Z1S301 9-11 Appendix A POLYCHLORINATED* DIBENZ0-O-0I0XIN CONGENERS 8Y homologue ANO ISOMER (NUMBER OF ISOMERS)** i Tot Tor e4 NumOering system for congeners MONOCHLOftO-(2) 1- 2- 0ICHL0R0-U0) 1,2- 1,3- 1,41,6- 1,7- 1,81,9- 2,3- 2,72,8TRICHLORO--{14) 1,2.3- 1,2,4.1,2,6- 1.2,71,2,8- 1,2.91,3,6- 1,3,71,3,8- 1,3,91.4,6- 1,4,71,7,8- 2,3,7- TETRACHLORO-! 22) 1,2.3,4- 1,2,3,61,2,3,7- 1,2,3.81,2,3,9- 1,2,4,61,2.4,7- 1,2,4,81.2,4,9- 1.2,6,81,2,6,7- 1,2,7,81,2,6,9- 1.2,8.91,2,7,9- 1,3,6,91.3,6,8- 1,3,7,91.3,7,8- 1,4,7,81,4,6,9- 2,3,7,8- PENTACHLORO-( 14) 1,2,3.4,6- 1.2,3.4,71,2.3,6,7- 1,2,3.6.81.2,3,6,9- 1,2,3.7,81,2,4,7.9- 1,2,4,8,91,2,4,6,7- 1,2,4,6.81.2,4,6,9- 1,2.4.7,8 1,2,4,7,9- 1,2.4,8.9- HEXACHLOAO-aO) 1,2.3.4,6,71.2.3,4,6,81,2,3,4,6,91.2.3,4,7.91.2,3,6,7,81.2.3.6.7,91.2.3.6.8.91,2,3,7,8,91,2,4,6,7,91.2.4,6,8,9- HEPTACHL0R0-(2} 1,2,3,4,6.7,81.2,3,4,6,7,9- OCTACHLORO-(l) 1,2.3,4,6,7,8,9 * EPA includes monochlorinated compounds under the general term, 'polychlorinated ** 'Congener* is a general tens used to wan any isoeer of any homologue; thus, there are a total of 75 PCOO congeners. In the present document, 'homologue' < defined as a 9*0110 of isoeers having a specific nuafeer of chlorine atoms; thus, the tetrachloro- homologue has 22 isomers. An 'isomer" is defined By the numerical arrangeoent of chlorine atoms within the homologue. HONS 21S302 A-1 polychlorinateo oibenzofuhan congeners 9Y hqmologue ANO ISWCR- (NUHSER OF tSOfCRS) t * Numbering system for congeners MONOCHLORO-(4) 1- 2- 3- 4- OICHLORO-C16) 1,2* 1,3- 1.4 2,3- 2.4- 3.4 1,6- 1.7- 1.8 1,9- 2,6- 2,7 2,8- 3,6- 3,7 4,6- TRICHLORO-{28) 1.2,3- 1,2.4 1,3,4- 2,3.4 1.2,6- 1,2,7 1,2,8- 1,2,9 1,3.6- 1,3,7 1,3,8- 1,3.9 1.4,6- 1,4,7 1,4,8- 1,4,9 2.3,6- 2,3,7 2,3,8- 1,7,8 2.4,6- 2.4,7 2.4,8- 1,6,8 3.4,6- 3,4,7 2,6.7- 1,6,7 TETRACHLQRO-( 38} L,2,3,4- t.2.3,6- 1.2.3,7- 1,2,3,8 1.2.3,9- 1,2.4,6 1,2,4.7- 1,2,4,8 1,2,4,9- 1,3,4,6 1,3,4,7- 1,3,4,8 1.3,4.9- 2,3,4,6 2,3.4,7- 2,3,4,8 1,6,7,8- 1.2,6,7 1.2.6,8- 1,2,6.9 1,2,7.8- 1,2,7,9 1.2,8,9- 1.3,6,7 1,3,6,8- 1.3,6,9 1,3,7,8- 1,3,7,9 1,4,6.7- 1,4,6,8 1,4,6,9- 1,4,7,8 2.4,6.7- 2,4,6,8 3,4,..7- 2,3,6,7 2,3,6.8- 2,3,7,8- PENTACHLORO-{ 28) 1. 2,3.4,6- 1,2,3,4,71. 2,3,4,8- 1.2.3.4.91, 2,3,6,7- 1,2,3,6.S1, 2.3.6,9- 1.2,3,7,81. 2.3.7,9- 1.2.3.8.91. 2,3,6,7- 1,2,3.6,81. 2,4,6,9- 1,2,4,7,81, 2.4.7,9- 1.2,4,8,91. 2,4.6,7- 1,2.4,6,81, 3,4,6,9- 1,3,4,7.81, 3.4,7,9- 1.3.4.8.92. 3,4,6,7- 2.3.4,6.81. 4,6.7,8- 2.3.4,7,81. 3,6.7,8- 1,2,6,7,8- HEXACHL0R0-' 16) 1.2.3.4.6.9- 1,2.3.4.6,3 ,31.2.3.4.6.7- 1,2.3.4.7 1.2.3.478.9- l,t,3,6.7,s 1.2.3.7.8.9- 1,2,3.6,7.9 1.2.3.6.8.9- 1,2,4.6,7,3 1.2.4.6.7.9- 1,2,4,6,3.9 1.3.4.6.7.8- 1.3.4,6,7.9 2.3.4.6.7.8- 1,2,3,4,7,9 hEPTACHL0R0-{4) 1.2,3,4,6,7,8 1,2,3,4,6,7,9 1,2,3,4,6,8,9 1,2,3.4, 7.8,9- OCTACHLORO-{1) 1,2,3,4,6,7,8.9- NOMS 215303 A-2 Appendix 3 POTENCY OP HAlOGENATEO BIPHENYLS IN INOUCING HEPATIC ARYL HYOROCARBON HYOROxYLASE ACTtVtTY IN CHtCKEN EMBRYOS Chicken ettCryos at 17 days of gestation were administered halogenated siphenyl com pounds dissolved m 25 ul of -dioxane, Twenty-four hours later, they were tilled, and their livers were assayed for aryl hydrocarbon hydroxylase activity, EO "50 (moi/egg) 'EDe ^50 {mol/egg) l eng- THACtlVC *.4 x 10 -9 inactive 10 n^Q)-<Q)-a 4.7 X 10 -9 inactive inactive i nactive 5 inactive inactive Inactive 15 <0)--(Q)"" intct1ve a <QHS> 1nict1 16 8-1 <QHQ> ,hiCt,v* HONS 215304 Appendix c TRANSFORMS DATA FOR TESTS CONDUCTED BY CHITTIM, ET AL, (75) Sample Installation Group 1 1-1 1-2 1-3 1-4 1-5 1-6 1-7 1-8 1-9 1-10 1-11 1957 1961 1963 1963 1963 1965 1970 1970 1971 1974 ICU Group 2 2-1 2-2 2-3 2-4 2-5 2-6 2-7 2-8 1967 1967 1967 1967 1965 1965 1965 1965 Group 3 3-1 3-2 3-3 3-4 3-5 3-6 1967 1967 1967 1967 1967 1967 Group 4 4-1 4-2 4-3 4-4 4-5 4-6 4-7 1947 1947 1966 1973 NEW NEW NEW Manufacturer* FP FP FP FP FP FP FP FP FP FP FP CGE CGE CGE CGE FP FP FP FP CGE CGE W w FP FP CGE CGE AO HA CGE w FP Olelectric Strength Otv) Power (ItVA) Volume (liters) kVA/1 36 1,500 1,888 0.79 36 1.500 2.093 0.72 36 1,500 1.251 1.19 36 1,500 1.251 1.19 36 1.500 1.274 1.17 29 1,500 1.151 1.30 30 1,500 1.765 0.B5 22 1,500 1.490 1.01 28 1,500 1.490 1.01 28 1.500 1.490 1.01 - 1.500 - 33 500 725 0.69 31 1.000 938 1.07 36 1,000 770 1.30 36 1.866 1.268 1.47 36 700 660 1.06 36 1,500 1.251 1.20 36 2,500 2,084 1.20 36 2,500 1,092 2.29 33 500 725 0.69 41 1,000 770 1.30 41 750 883 0.85 41 1,200 1.087 1.10 31 750 829 0.90 36 1.000 838 1.19 56 10,00.0 m m - 660 6.900 0.08 1.45 m m - * FP - Ferranti -Packard CGE - Canadian General Electric H - Hast inchouse AO - Broun Boverl MA - Foster Electric C-l HONS 215305 Figure C-l. Fragmentogram (Based on Ness Specific Detection) of e Fly Am Sample (Above) and Standard Mixture of TCDO (Bottom) on a 55- SHar 10-C Column (l.d. 0.26 mm) (113) C-2 MONS 215306  nr>m lll-li: *4. at it. > -*4.at m1 I m X Flour* C-Z. Riltlpl* Saltctad Ion **ss ChroMtopraa Obtain** for Calibration Mixture Containing th* ZZ TCOO Isoa*rs (Colmat: 50a Hybrid Ph*** MCOT Slats Capillary) (Braha Laboratory, Personal Comm Icat Ions) MONS 215307 C-3 -3 (1) ).i.ft-t#lr*-COO <11 i.M'ltir* COO 01 2 UJ-tilff-CDO 1*1 i..2.a-eti-t(io 01 2.1.?.l'ptiiU*C00 1*1 (1) (a) <> < 101 on <121 i.i.M.U iwu CDO (1)1 DeU-CBO ao 13 to 9 Figure C-3. Matt Fragaantograas (0V-1F Clast Capillary Coluaet, 200*210*0 Showing Elution of PCOOs In (a) Fly Ash of a ftntlclpal Incinerator, (b) Fly Ash of an Industrial Heating Facility, and (c) Hlcrapyrolysls Saaple of Combined 2.4.6-tri-. 2.3,4,6-tetra, and pentacftloraphenale; o/e 320. 354, 388, 422, and 458 for tetra* to octa*C00t (34) MONS 215308 Figure C-4. Ness Frtgaantogran (m/e 270. 304, tnd 338) of PyrolyZtd Pentichlorobi phenyls Shotring Elution of tri-. tetri-, end penti-CDFs fro* (t) 2,3.4.5,6-, (b) 2.4,5.2'.S'-, (c) 2,4,5.3* .4'-, (d) 2,4,6.2*.4'-, tnd (e) 2,3,6,2' .5' -pentichlorobi phenyl (50-ei 0V-17 Slits CapilUry Coluwi, 100-160* it 20*/*in, 160-250* it 2`C/nin) (34) C-5 MONS 215309 Flgur* C-5. Mtis Fragaantogran (/ 304, 338, and 372) of Pyrolyzad Hex*, chloroblphtnyls Showing Elution of tetr*>, pent*-, end hexe-CDFt fro* (*) Z.d.S.Z'.d-.S'-, (b) 2,4,6,2*,4',6'-, (c) Z.S.S.Z'.S*,*1-. (d) Z.S.t.Z'.l'.S1-, () Z.J.S.Z'.S'.S*. (f) 2.3,6.2',3',6'-, end (g) 3,4,5,3',4\S'-hmchlorobi- phonyl (S0- OV-17 suit ceplllery Co loan. 100-160* et 20*/a1n, 160-250* *t 2*C/1n) (34) MONS 215310 C-6 C-7 Figure C-6, Mass Fragaantogram (a/e 338, 372 , 406. and 440) of Pyrolyaed Hepta- and Octachloroblphenyls Showing Elution of pent*-, hexa-, hepta-, and octa<CDF froa (a) 2,3,4,5,2'. 3*. 4'-, (b) Z.S.a.S.Z'.a'.S'-hepU, and (e) 2,3,4,5,2,.3\4\S'-octach1orobiphenyl (50-a OV-17 Glass Capillary Col urn, 100-160* at 20*/tn, 160-250* at ?*C/1n) (34), MONS 215311 Appendix 0 EXTRACTION METHOOS FOR PCOO* AND PCDFs U.5. Environngntil Protection Agency ^EPAl Procedures (1531 Acid-Extinction Procedure Applied to Fish, Adi oote Tissue, Ml Ik, Miter, Soil, and Sediment-- Solke 10 to 20 g of sample with 37C14-2,3,7,8-TC00 standard, i Reflux with KOH. a Extract with hexane. e Remove and extract hexane fraction with concentrated HjSO^. a Dry on sodium carbonate. e Chromatograph on neutral alumina column using CC14 and CH^Cl2 t0 elute. Neutral Extraction Procedure - Applied to Fish-- e Slend 15 to 20 g of sample with sodium sulfate and dry Ice. e Spike resultant powder with 37Cl4-2,3,7,8-TCD0 standard, e Extract with acetonitrile. e Remove acetonitrile fraction and extract with acetonitrile-saturated hexane. -a a Discard hexane. Concentrate acetonitrile and replace with hexane. a Chromatograph on florisll column, then on neutral alumina column, eluting sequentially with 100 percent hexane, 10 percent CHjCIj-i"-hexane, and 25 percent CKjC^'ln-hexane. U.S. Food and Orug Adalnlstratlon (FDA) Procedure (176) - Applied to Chemical formulations and risn Dissolve 20 g of sample In alkaline solution by agitating at room temperature for 2 to 3 hours. a Extract with hexane. a Remove hexane and extract withconcentrated acid, a Dry on sodium carbonate. a Chromatograph on neutral alumina, eluting with 20 percent CCI4-In-hexane, then CH2CI 2< e HPIC on Zorbax-OOS, 40*C, methanol eluant. 0-1 MOMS 215312 U.S. fish and midlife Service Procedure (177) Blend sample with sodium sulfite to free-flowing powder, e Pick powder Into glass colour and extract with Ch^CIj (200 ml for eicn g of fish). e Evaporate solution to 50 ml. e Add methanol and benzene to yield solution containing 20 percent CHjOh and S percent benzene. - e Pass extract through colour containing segnents of potassium silicate and silica gel. e Pass solution through carbon-glass fiber sorbent, and wash sorbent wi tn CHjClj/CK^H/CjHg (75/20/5). e Elute with toluene. e Chromatograph on column containing h^S04 dispersed on silica gel and cesium. e Chromatograph on acid alumina, eluting with hexane and CH^Ci^-hexane. Dow Chemical Company Procedures Acid Extraction Procedure (176. 179) - Applied to Fish and Milk-- e Spike 10 to 20 g of sample with 13C-TCD0 Internal standard (and/or other PCOO standards). e Shake with concentrated HC1 for 1 hour. e Extract with hexane (overnight shaking plus 3 additional hours), a Pass hexane extract throu9h combined column of silica, concentrated H^so^ on silica, and 1M KDH on silica (l.e., 22 percent HjSQ^, 44 percent H^SO* on silica columns, and KaQH on silica coluams are used alternatively). e Pass hexane extract thrau9h second dual column of silver nitrate on silica and basic alumina. e Clean up dioxin fractions using normal phase silica (Zorbax-SlL) HPLC, then reverse phase HPLC (Zorbax-OOS, amthanol solvent). neutral Extraction Procedure (180) - Applied to Particulates (Fly Ash, Dust, Urban Particulates), Mwlclpal Sludge, and Soils-- o Spike SO mg to 100 g of sample wi th l3C-TC00, l3C-HC0D. and l3C-0CDD Internal standards. e Soxhlet extract sample with benzene for at least 16 hours, e Concentrate extract using Snyder column. HONS 215313  Subject to liquid chromatographic ano hrlw%c* tnit.f,tfup similar t described for tcid extraction procedure. t0 tha* Wright Stete University-Brehm Laboratory Procedures* Procedure for Soils, Cheetieel Wastes, and Aqueous Effluents (ISO. 1811.. Soi Is e Spike 1 to 10 g of soil with ^C1-TC00 Internal standard {and/or other internal standards). e Extract spiked soil in bottle with petroleum ether or hexane<*ethanol by shaking for period of 20 minutes to overnight. e Alternatively, extract spiked soil in a Soxhlet apparatus with methylene chloride or benzene, followed by concentration of extract on a Snyder column. e Wash the organic phase successively with 20 percent KOH, water, concentrated HjSOa, and water, discarding the wash solutions. e Dry organic extract over sodium sulfate. e Pass extract through basic alumna column. e Clean up dioxin fractions, using normal phase silica {Zorbax-Stl) and reverse phase (Zorbax-ODS) HPIC. Chemical Wastes e Spike 1 to 10 g of sample with ^Cl.TCDO internal standard (and/or other label led standards). e Extract spiked sample in bottle with methanol and petroleum ether by shaking for period of 20 mnutes to overnight. e Continue with cleanup as in soils. Aqueous Effluents e Spike 10 to 20 g of sample with ^7C1-TCDD internal standard (and/or other internal standards). Extract spiked sample in bottle with petroleum ether by shaking for period of 20 minutes. Continue with cleanup as in soils. HONS 21531* 0-3 nd Other Simples from Flue Gas Stack Sampling Trains procedure fr p<rticuite* (160. JK)** Solk* appropriate quantities of organic and aqueous liquids, particu lates. XAD-Z resins, and other sorbents with 37C1-label led and/or 13C-labe)led PCDOs/PCOFs. e Extract liquid samples with petroleum ether by shaking m bottle for 1 hour. e Extract solid samples in a Soxhlet apparatus with benne. e Idash extracts successively with kOH, water, and and discard washings. e Pass extracts through a combination macrocolumn of silica, 33 percent NaOH-slltca, 44 percent HjSOa-siHca and silica, eluting with hexane, e Pass eluate through a Uoelm basic alumina xrlnicolumn, eluting with 3 percent CHjCIj.in-hexane and SO percent CHjC^-m-hexane, collecting appropriate fractions. Umea University (Sweden) Procedures Procedure for PCB Cleanup {10)-- e Chromatograph 100 mg used PCB on an alumina microcotumn (1.0 g basic alumina, Uoelm, in Pasteur pipette), eluting with 10 ml of n-hexane. e Recover PCOFs by eluting with 6 ml of CHjClj after careful evaporation to dryness. e Chromatograph on a second alumina microcolumn, elutinq with 10 ml of * percent CH^Ct2 in n-hexane. e Recover PCOFs and other chlorinated tricylic aromatic conoounds with 10 ml of 50 percent CH^Cl^ 'n n-hexane. Procedure for Fly Ash, Soot, ulpe Tests (^, 183. 184)-- e Spike 1 g sample with 1 ppb of Z,3,7,8-*3Cij-tetre-CD0, Z.3,7.8-^C14tetra-COF, and 37C1g-octa-COD. e Shake for 1 hour with 10 ml of 1M NCI. e Filter by suction using Buchner funnel, and wash with water. e Dry in air, and extract the dried material with approximately 100 ml toluene In a Soxhlet extractor for 48 hours with a speed of 1 to Z ml/min. e Reduce toluene to 100 ul by evaporation on a rotavapor using some external heating. HONS 215315 0-4  Eliminate remaining toluene by a Purified Nj flush -1thout external heating. Dissolve th* residue in 2 ml of n-hexane. e Chromatograph on t silica gel column {0.5 g Kiselgel 60, 70 to 230 nsh (Merck)), eluting with 5 ml of n-hexane. e Seduce n-hexane under to 2 ml by evaporation, e Analyte for PCBPs. ~ e Separate PCODs and PCOFs us mo anAlox column (1.0 basic Aloe akt 1, Woelffl, 1CN West Germany). e Discard the first fraction, i e., 10ml of n-hexane:CH^Clj (98:2). e Collect the second fraction,i.e., 10 ml of n-hexane: CH^Cl2 (1:1). and evaporate to dryness under Nj. e Dissolve the residue m 100 ul of solvent. Procedure for Blood Sample (_79. 13S)-- e Add 10 ml of n-hexane to 10 ml of blood plasma, e Spike with 0.1 ng of ^l2*23,7,8-tetra-CDO. e Add 10 ml of acetonitrile saturated with n-hexane (three times), e Shake mixture and centrifuge. e Treat the caetoined acetonitrile phases with 10 ml of 1 percent aqueous sodium sulfate solution. e Extract three times with 20 ml of n-hexane. e Ory the combined n-hexane layers with sodium sulfate, and concentrate to 2 ml. e Chromatooraph on alumina column (1 g alumina). e Elute first with 10 ml of 2 percent CH2Clz m n-hexane, and discard the eluate; then elute with 10 ml of 50 percent CH^Clj in n-hexane. e Concentrate the eluate. HONS 215316 0-5 Appendix E GLOSSARY AMO ABBREVIATIONS AHH ala Aroclor Askare! Ctrl ortent Congener Dioxin Edna Frogmentogr am Fur an Monologue In vitro In vivo Isomer Kanechlor PCS PC BP Aryl hydrocarbon hydroxylase; an enzyme. 4 aminolevulinic acid. Monsanto trademark for PCBs. A generic descriotiive name for synthetic dielectric material; for the purpose of this report, usually a mixture of PCBs and chlori nated benzenes. An acne-like dermatological disorder due to the exposure to chlo rine compounds. A general term used to mean any isomer of any homologue. Otbenio-p-dioxtn, a triple-ring organic compound consisting of two bente rings connected by a third ring through a pair of oxygen atoms; in popular parlance, dioxin is used for the 2,3,7,8-tetrachloro isoamr. An abnormal accumulation of fluid in the body. The term used for the output from a mass specific detector miss spectrometer. Specifically, a five-sided aromatic hydrocarbon with four carbon atoms and one oxygen atom In the ring; in general, the term is used for the dlbentofuran and its chlorinated isomers. A group of 1 soamrs having a specific nuatoer of chlorine atoms: thus, the 22 tetrachloro isoamrs would be the tetrachloro-nomologue. Outside the living body and in an artificial environment. In the living body of a plant or animal. One of two or more molecules having the same nuafter and kind of atoms, but differing in respect to the arrangement or configura tion of the atom. Japanese trademark for PCBs. Polychlorinated biphenyl. Polychlorinated blphenylene. HONS 215317 E-l peer Polychlorineted chrysene. PCOO Polychlorinated dibento-p-dioxi n ; Oioxin, PCOF Polychlorinated dibenzofuran; Fyran, PCDPE pepr Polychlorinated diphenyl ether. Polychlorinated pyrene. PCQ PCQE Polychlorinated quarterphenyl. Polychlorinated quarterphenyl ethers. PCT Polychlorinated terphenyi. PCX Photolysis Polychlorinated xanthene. Oecomoosition of a compound into simpler units as a result of absorbing radiation. Polychlorinated For the purposes of this report, substituted with one or more chlorine atoms. Precursor Predioxin An intermediate compound in the synthesis of a specific substance; a substance from which another substance is formed. A diphenyl ether compound with a chlorine atom and hydroxyl group positioned such that they can react and form an oxygen bond be* tween the two rings. TCOO TCP UV Wipe Test Te tr achi orod 1 be nio-p-di oxi n. Trlchl orophenol. Ultraviolet radiation.. A sampllno method where a specific area is wiped down and the wip ing materials analyzed. Tusho pice oil disease. HONS 215316 E-2 Appendix F MATRIX OF REFERENCES FOR PCBs HONS 215319 Mir*ik of KftatNcft fat *et* _ tffWtttl. Ill- fUt- om cr<a */i tin* mtr in* nrt MMrtfTt MM MIN CCI tMV rt*i r*r/ Kli HIM Ml NCi CM** CNtN CMf CHCM CHKH CNfN CUM MB VM TM I NCR HPT (NT* bit* *MtC l .ft M MNTlH. I TV M ||IT| MTicn riM in ft it? in rue *t*.NTIM IIVIM IN TNt fCtt NUVTft MU IT MVSM INi CMtttlNftftt 11 OK INI MM Bitblll CMTMNII, IBMCt m im CNVINMMCNT t NNTtlMtft. If M .Ill HIIMM IMtt.lLBIMai. i T Jilt ** ITI-lfT NM *circtHM.t t .cr in Mtr***iNt cut#- I CM. NttTKt TN fiMKKIM tCNLK INClNtMTMt rtCHlTV tf*Mf Nt I. ItLkINI ItfVltMMfNTNL tttVICf* t(M imi. riMi tm Mrente mm mkc tvtrcNi mmi.iimbm mkb.cn.if , III?. ir * mM KtibMH.i t *f t n Mitvttntt *** TNt ITtMMt if *Ctt MM K IftNt V INfMMt tatTMCfltN t**-tl|/'tt-U> .TM.NNBM KNCNfCNLI* .MtMMM INI x k n NCIMtl CtftfttAMtN IlMUN M *tLTCNLtNINNTft limimt (KMI MM *CtCtNfMHNNTtt NtTCtlM.*. ML. 4 fill iNCIMMTltN M tUCTtlCM. CNfttlftNt CtNTOINIM *CM 1**1 f*-Utf, CUCft 1C FMI NttMU INtTITVTC. **. NLTt.CM.If .WtftNttt IKI. N MMMtl,* .T NL MMLTtlt * *<TCrRNON*MtlMN2fl-*N**-tltNIN IN TIM HVIU MM* IN* CNLtNINNTtt Mi MKT MINI MM INC |fl tlWINt C MML.lt CCNL NULL < *?tCNN*LN1.** tl-N.1t** 1 M n HONS 215320 MtllM 9$ KriNMKt FN Kit an- rutkm Hint am 9111 cHfn t/t nv mT m dm nh acta net cm** CHfN (Hr* CHfN CMC* cm* cm* cm* -- mhh atca mv tt*t tat/ tm tin mea IWT 1 -- tur urttraci MLTM.i MltHttrli V CMLM HATit I I MTtttCitifI t* CfM*T Ktl* IWtMt KI.TIOMI UK ' _ M tMLiM.a * LWltH aaiiiiM aaaa m tMLitlMITtt MMtC MMtMCII r-- Mamrrw n< cmlmImhi tit- it rat MLam cmhmh.immt n TM CM--ft. I. MTItMUMf 9L,. KM KMMM FattC.tL--pN.Y., IM. M.Cl-*. a tMLIM.t fMI I.UNKN. IM--4 teat-- tim it ki mi mmatmmi a--. Mi.miL --i-- .titt*. rtf*. I K LlM.t .ft IL, if--TIM ar MT* MLMIMTM JtNMMjjllMril Ml^ll- s WMM.I MitKMlICIi MMTI-- ar pm--? Mtaa -- iimm. ta* MMiMm ammwf mim -- tmip aiMiM. c.--tL.tt. ceai mh.l. < tTac--ui k ar* n-ai. ran. it--mu.a *--tmmimtim ar M--MTIC CM--. Ill aiMla. -- ICaLMICM. -- CHftlCM. MPtCTt. r.CAff--*|,t>*PtLLM*.Mt ( Mill tm rr.tti*tai. II n ft k a HONS 215321 AAtftlV IT IKFCKHCCI fM KCt tmwnci 10- Mli AlNl TN CMN 0/T TtW FNOT (H HfC MM. AC TO mm AMIR ACC* fMV r*Nt Til/ Ml CUM CNfA (NCR CHfR CRfR CNfA CNfR CMCM FOR TiR IMI IRRT I MFC I IF MMI.I M .MR 4 CMOtrt CltlM' MM CMC CLKRM-UP OF CM IMM. tlMMI FM tMMOUtAT OtfCtMHMT IM Of MM- tumc or CNiMiMcrto oiot*o r-oiw- INC CMC HMMWMCMt. CttC^MANKM -11 Mi-m. nrr M* MR - almo.f v .rmc c c amuo cmfasi- i i i M OF TMf CMOMIflMM IF CMClM t|4| CMC CMCtM lilO J CRMURI . tOtiUl-Jii.IWi ALOAO.F V .UNO C C NMCO. ttMMt AFFMACM fO INC FOACf IAMMT |M CMC CLM OCTftMlMCMM OT CMMMLfl M|K- TUMI Of (NLNIMTtl MMMMC CMFOUMOt 4 CMAOAAftOO , 107 JI SO-tOO, IMS ALLIN.J.O .AMO i.R.MQMACr FOTHOOIO- LMrCAL MlFWtfl or PIIAATfO ff FOLTtALM tAATKO OIFMMYL tWFOMOt Tit CUfCNDCK FOOCttOlNftl* ACTIONAL CAMKOCMK M FOLVtNLMlAAfCO or- RMfRVLl (HOVtAMO l*?0, CRTCAOO. LLIMOTik (FI-UI/l-H-tM NCACM tore FA IMI, 4ALLKR 0 ,IT AL AOMNOLMICAL CMCM- Uo IN AMCVt COAMAIM 0 OIK1 CON TAININC LON LCVfLO or o.j.f.o-titm- 411-41CRLMOO tOCNM-F-CIMlH FMI COONCT TOHICM . tit , TOFF IfATCAM CNfRTCAi OOCHTV AMIMLINKO F00 M?A ACOAIOTTIAA ANA MfA MALI TV OVAL NAT TOA IN <NV|OOinCATAL CNCNIfT<V ANAL.CAIN ,10*0040-0041. INI III N * \ n K * K HONS 215322 Mfctv r acFtiKhcn fn kii MftMHCK 19- FM- 9IN CNfN 9/1 MMT INV NFC MNH 9CF9 NUN Mil CC ftfv tM* Iff/ FC9 MINI NN9 HCl CUM CNfN CNfN CNfN CNfN CNfN CNCN CNCN T9N F9N INC9 IRH INFO 9IFF AnfRICM HfCtCM. HtMlartM tNf NfftLTN IFflCTt W ftNl aiMIII* MM NLycmwtiMfM nionin CMtiriM(ri. CMJCM9.9CT9M9 *OL?l * MltUNM FMFa'4IM 9F CJN.MN9T- V It cvHLiMfian in mmfttMiii - mt- IN MM rl| C N M CMC# 9Cf..lttIM-l.19?l. 9ttwut umc ifmt fiarf-tr- rm-iir or nimin fim cmmitIm IlMCfl NNffTCMI MClIfV 9f NfCHMII- CM. iMliilM.Ml 9 9 99 imim * ,t? m. imtflMi m mi 9NlFallC UlfTaTliaie*l-tftMlFltM *C- rtvinc* * ,>. -f?cm--n>ifniM-irurH ritoeiaiMtii irwin MM mTK CMVMiMt. HM*Ct Ml tHC IMIMMKHT 9 MIIIMII (T M.. ,t FfMMMM F9f9f.KLN9Mf*N.Y ,f90t rr.it!-49i 9 9 NtfMMMI.9 .MM MNLMN Ml IMf* VC9 MMCT9I9 FM TimcNiMMlHilM* F-ltMIM. INt (NtMMlUtM - MICtN MM F*|t f N 9t*J.C9 N99 CNfN Mb ttlttl-lM.ltFI 9 9 MMNNMl.i 9 TC99 MM UMV9I9IM. NC~ CTMNT9 Ml 9 9t9*9MTIMI.MMVMM imivc9itv.v?4 meant* r i UNlVItlM INC NCNMLIM MM INC TtVIC CIOIM TUI NMIVC99ITV MMfS.NfN NN* VCN.C9MN .!*> M HMW 9 99 HONS 216323 **ru m MriHMCKi rw mii #i#- ra Mil imm MHY.l r M* MVt T* MITNV MCM. CM* IM .MtU)>-4l,IMI MlMMM.F 4 ,* C UMLIItM MO M.MMCiV. MCLvtMtMiMTC* nncimi mmm MirafC IIMM. MU MVtMB.CMTMf. FMMIC flillMIMin LMMM.i i ,CT ML. HTINMTrCi IT CM-MIMIfi MMHM-^-ilMlM |i PtmTMCMMMMMi >V 1*1 CNMMTMMrav Mil PaCtMMTtV MNflL CM*.*MttM4m* tm MLTM.t MMM UMfM* M MLiflM I* IIMI. CM* CM..M<ttiPIMri. MtlCM.i M .t| ML. IMCflli W i-PCtf*n*M MctlviTV m mm tmtx r# IMMH TM CM-Mtmi MiWII MMM TM*CI f*9IM*M*IML raLL*T*MTM CLI* CMM ,lhllll-l||I.IM mtm,c .m mmli^mm r.ml-mmimm MM TM MMMMMMM W t,S,?,*+1fiMCMLMMMMIMMM-MMMMIMMlM (fM M MMM M M Mi CLMMM MT t*LfMIMMIMCM CMilCMM Mf TtCMML ,!) m-iiMin MTMI.C - * MMLI*MM I ML-MMtMM TCW tMLMTLICftllMM MM MMTMMCttraCTTICM IM MMMMMM NLiTlCM IHVtMM*.MCI TKCMML , Ifjll-IM, IMP* tMTlTil.M T .MMM M MMMMiTf. MCM*NIMMM FN tlMMIM NMC MVMMMLVMCM MM CMC.MMMiHUIM M> NNLMIMLMMM 4.MM CMM CMC ,ritlM-i4MIr, tM CMM M/T fM rail IMV NT6 MMi MCFft MM MTU MCI CMV Uif 'it' w CUM Mil curat OK* CMM CM* CM* CM* TM F|M INC* IMMl iMf ll HONS 215324 MkTCTJI w |r|H*C|J ft* io- riw- m chcn i/f nw nit cm arc mum. mi* NVH MtM MCI IMV tt*l t#T/ mu M*+tt* *Ct tIM* CNti tWN CNtN (MH cmm cmi cmn m ?* inci i*r* ft's* NMCM H (T M MO CNlMrNM' OMTC CUMMTIIIItKl C OVtCIHIC cNLoorMtto iMNiaruMti Ittimrrmum c tm racMit t mciicm mm jMMMM MLVamilMIII IthOMlI j oootc.ow taMB.*cj\ria-im,itn I 4 NMM M .Ilf M. iMNttfiCOftM IT MLNIMTC9 IfMllMNiaMi IN MiaiCM MLVCMLMIMTIt MUNI. IMilN'IIMm * HMI.I II.,ft M. IMtflIIMI Ml rKO* -n rriiciriM c (NLMiMltl tiMMt* . fnmi ratal NmcnrMTci itrami ** cFC> MM rtM WMM ltd tK CM* TftiNiM# oca. iii iiMiit wrcciilcat MM CWI1ML MMCTI r.ttttftMIM MMUM,Ml c MUr.lH PCCTMM MUMtlM.lMI fMMlN pp m-m mmm.* ,mm o.t curmrii imcriM m nmK acmirr m mu mTMCI IMII UMtR m KflCrlM m MW MLaCCMIllMII MIMIC CMMMMM 4 MMC IM MMi.CflfB 4fi IM-IIMIH R > liniMU.I C. MS MM MliMIMkl* IT r MifCIMIIMTD TMMlI. KimaMMltTI M>M|t.,nHIIMIim IMM CilMM.im, m*m,4,rttm mm mol ft cfMCTi r vtcctaicat mum rct c. bcotm (iterate kkmcr mm mmimmit CClTK,fCCCCfMV,l T , INI on* > HK ** gmr H HONS 215325 **r*]k cr (nuNCKi fm kk irmuci MWM Nf mcNTNC HUM Ml INTI MLf HUN (NIMMMI), N If. IMS li- (ll Hint MUNLKf.N C pit M LM-CCMtiflM NMLTIEtR IM MTICTIM Ml IWM TMMMlC-NNiS WfCfMNKMIC CMMIJMN M tM ClTNftCTIttt'CLCMaW TMI EM MfSNNlNIM M.M-TinmUlMilMNSi-E-ilMlN IN TIM J.NMIC. EM* CNCN pllilMHlMI.IIII M MM I I ,KT N TMCC OKNIlfllll m mElMi inki cnlmimtm SIMM*. ttfllieUllMHlilM*lllr Cf HJMI Mill I 4 . m NMMIK CNNEIfttNCt M II MNSMtMIM tm llllWNNMH, Mt INMi. IfffIlNli W IWlMMitiML NfMfN NWICII, Mill S-I IMS. CMIMa NCNLTN NCMNfC PfNNKCtttCMTM. IIM IjITHNJIM MIM I MN.IIII m NIMN.MIMtll ilMNtt-E-ilMlM tm IIMHMEMNM IN CMMllMTCi NHCNK.I It NMI ENMMNTMiMNT J CNMMfM* . mh-ki, im N Mttl.lt I fVML.fiII 9P TCM i if NM CUNWTMiNNNt-nMi MtCTMMTNT MINK ILVII CWtLLMT CflLMNN lit INKINi TiNlCCLKttM N CNfNlCM MMCTI E CNIINMNIpf CNVNLLNM.NN* CM.LI.KiS MtCtNNN MMItNTlKMr NKN VMv.li'i PP E-SI 1 IN CttKN '! few NNI INV we MML t| Nun WIN NCCft ENV INNS TNI/ Kt CUM CNKN MU CNf CNfN CNCN COIN CNCN tM ?* MC tNNT INTO SI*F K I II K* K HONS 215326 MltJtf If HFIHKh Fit tCM ittiimci nni n MfiMiMiriM m M.r, l-tltlMifMOIlMWI-t-llMlIt IN KNVIMNNKNIm. IMMI IV NIM-NNUtlNl Ml OHImIMMNV Ml LM NfMLMtlNN MM VfCTHNITiV MHH CNfN, NiiJi-MMir? 111- fintCll Mli Ml IN CNfN / TIN Ml fNV NV MML NfT> C CUM CNfN CNfN CNfN CNfN CNtN CNfN CNfN WN MIN MCI fNV TNNI INI/ IN IH IMI IfMt lff| ij$* MMI.I 1 MMhTIM NN iNCNTtVIC*TIM m HIM* Ml fHINC--.ft HfHM-f-itMlINI MM NNTNVIII W TM IMNSNIC NlfUMNINW-l- tMIIM. ClfMINm,ltm-MW MUM I fMMTIMl if MIVULNINNltt IftWnMUWl tKBMl Ml BfKM-f-flMINf t PC>111 MM TV MMtmi m CNLt MN|IIpI<III-I|I. HN. MIM >. NlfN MUtltTM Ml CNN** MTMMMV m INf it flTMCNJam* mtM-IIMli rtCMI ]MMR9 in tNLHtlMTM IMIM Ml MlltN CMMWil. INMCT M TNf CNNlMNHKNT >.NNT)INtff,fI ft ,tl HUMP MtH.llMMM.I V.,IM> NN It-ff MTMMMV m NimLMIMni IIMN* ipmimiin mm iwnmwmm mu. CNfN .NilHMMMtn, MUI,H MlVCNLMlMm IIMIn-MiiviiN mm iBtanrNNMMtrim riM.UCMMMfl MM NMM.V>I> M >NVIHNNfNtM.LV NtlHIMM CMMNM n > nifiii hmvihitv m mm* mm, MCHMUI HONS 21532? MTtiii ar tcrmiicca rat -- mcm a. tatvcaLaaiaavca araca- *a*r*ata*raa* tcManaa aaa lacattrtcartaa ar itana* av taa entanfa* caatav-ataa arfCTaaacfav j fanaa rata ,tMva-iac.itra, Kl* Kf a mmi.b a. aattaaaTiaa ar aaumtvi iTaaaaat aivraaaa ar racvataaraaTca atacwca-a-ataaraa aaa aiatNMwaat av utiaavtatcf aaa aaaaa laaaaratraa ar tac acracacaaa caraaH j.caaaaaraat . uiiaa-iar, im a nni.ii a <aaa c aarrc raaaarraa ar r&TtitMtasTu artCBMraaaaa caearai raaa rac hfaa,Tir ar imviaai ki laaaua cataaaracif.attar*rtra a met a a., aaa c. aarre tia wmr* riav cat ewaataatarm ar rac n raacai.--aaiacart-r*aiaata laaacat. aaat rata ,cc icaff'CMt, raaa a aaiM,t.,a c.aarre raaaTrrTca* riaa ar amaTTfatiaa aamaaa in aLvcia.aaiaaTca aiatam-a-tiaaiaa ircaaat tv mm caccraaaittv caoa* tram.cita-aii.itra a aaaaa,a. .aaa a a aaaanaaat. acTca* ataatiaa ar a.v.r.a-Tcntacta.aaaai!*?-* iaaia av tatva act aiLtiaa umi ta maaicai aaaac ti.t-nr* tammmmmtaccTic acia ia m.i*i aLRVL tam aaa s,4.a-T aavai mt Ncaaictac raaaataTiam av aammu mi raaaaaaraaaarm j.cmtTm * a * a tv cm* a/T nv mt cm arc mml rc aaa ata acca cm raaa rat/ ac cum cata can cata caca caca caca caca taa laa face imt iarc att* a aa aa a a a HONS 215328 Mm* m itrtHMti rat kii ara- ri' bin tarn i/r nw pmr fw arc mm. n mm wra acc* *av icaa rar/ PCBl mi aa Ka cm* chcm cm* CNfH cncn tmn cHtM cmm tik th I IKa imt rara cn* .m i .am n a miiimmt ntu- NrtMTrto F MMYCMLMfMlVCa IINUtlr-aiaariaa tm iiwiiwmk in caaNtBCiat MNTNCmMMuml 9t cbmihta ui eiMMraa**inf-Mi mktmNfTftt * *.*?.MM.. ua-wa.iiH vcia.a iM n,m MtfMtr tatrCNLBtJCBfC IMNM-P-tJMIMt. IINH* nflMf M MlMil IN ni MCnc IMUMLII M IRMTIIKILII HI* aacanaaciaia an nut.mb lm' NiiiCLWicii.aarfl n a..c aam.aM mm. mtOMiiMm aiaanWM <kM) pmm in vmmm nil tm in ma **- NCM MB. CNtMMMBtrTi4M-44B,mB BWM0.N .* M. iMNTIPICNfltN N aurCKMIMTH IMNMrMHM IMNC10 rn nt MN UN NCM avMLYMI CNCMi,ri4rMiil tm nnmn.m a .a.-a MNimi.M c * M PMNNflM m MtMIMIMTO BJnawfNaai cmbfni fMH tm nmitaurtmt mbb. nowMan.Mi^ia, .n ,n -a mumbi.m c nmpi iBCMiTicavtaai a* mteNiaiiMTii NiacNM-a-arNiN imkh pbmn in fiv mm tNCMMMtM.TiHN-aTt.IBTa. cmmi i tir at tNNMafatT lavtairaafttN rat me aicaMratat icat acuta - aafttN at .j,?.a-Tf TtNcatMNftfacN- la-r-aiaata in nh av aaNiriaa at aacaric tfreer tM. caicara*Tea aimim m m*ria caarMNaa. rwacr an tnc cNvraaNNCNi a Mmacia.fi at ,aaa atauaM Mtaa.atNMMa.N w taat pp aa-iea HONS Z15329 Kirin or RirutMii rot kh JSU1CIKL CMIMI 1 IT M. IMOUCnOH OF NlftfO FUNCTION OXTMftt tv CMKOMrt TNfOTNfNT mitn a i r.o-rtToocmoto-orotHtO'Oi0*IM IN FfftMi Mil IlMCOlNrJTl is*-i*r root oio- noFCia ! Ml N Tn chcn o/t tint mmt inv ore m otio NCf CN*0 CNfN CMH CNfN CWH CNfft CNfN COCK * ' MM MMM 4CC0 f MV FtHI INT/1 m tod inco inft info omf CMiiiiir^in * ,ft ol rot OCIOC- ortioh mo function tnc cttomlic OfCfFTOt rot a, J.F,0-VfT0NCNL0NMOl- ocnjo-f-iionin <rcoo> no NtLorto co- COOCINOUNO l> CNLOOINOTf0 OlONfMt MMO ifLOffO CMNNIH, INFOCT ON TNf rovrooNNCNf o Mrniwi fr n ,cm FftOOMON FOfii CLNOFOOO.N V TOOO rr ns-us, n cmmii v owriu or rue mini MNICM OCCUMO IN 00Of00. IN' OlONfN TONTCNLOOfCM MMO CNfNlCM OTtCTi r COTTONONI.* CMIUNM.NO o.om.lt tit OFtCTtUM FMUCOTfOMO.NfU TOOK . till FF i-7 eMMLMl tIT IL NfTMOO FOO TNf oorcoNTNOTiON or t.. f.o-tktmcmlooo- OlitNM-r-Ol0NlN Of F F T, LfTflt |N VfOOTOOLOO it NlCN-OfOOLUTION 000 cNtiNo rociorm no lon-fo mlhtion Mil OFfCTOONfTOY CNfNOOrMfOf.il iai-iii.itoi CIVKUMM.I .fr M NfOOTIVf ION CNf NlCM lONiaoriON Mi Of 0 tTOW TOM TOOL TN TNf OfTftNIMNllON OF OMLL ornounto fcoo mo rctr tn. cmooi NOTCO ilitrlNO MMO OfLiTfO CillFBWHi, INFOCT ON TNf OlfVfOOMMtMT a NHTfTN- Cfi.fT M .000 OfOOORON FOfii,OUM- FOtO N r .TOOO FF 00-40 I n X MOHS 215330 mix or fit tcii ^ucitmu_______________________ CIVM.IMI,* .|f M. tMtllM. KCW. ICNCI, Ml IWiMIIM If KIM IK KKI KM IWlMtlfll ICUl MBM Mr* CM NWMfNt, *1*1 t-W. tMl CMM N .If K CMNMritl MIC! M clininniim m mm fftftrviMMi NLfeiHIIIMttl UWimi fM tM MM Of Kl-KTIMfl H1IIHTI IN MtlM n M KMIMI I* MM CMN V*X|CIL fMN.F m ,tr L LCflLI MM Mf CMI' MUMimic nitviii m mlvcnlm- tMici iiNiani i in km m m* rictiii irici ki NUMrii in iiiimn MLi twill* an* mjm Cl*IAN VMICIl , .< CMtM Ml JlK MTMHlIHTtl II- FMmi, IIIfNMMlW# Ml mttfll - Mttm.1 m w fMic iicc-mm iil mm ten I* VMt KM m Ml ICIVl I If* KH KttWlM in TitMM i II M NK1M8I CNffMfI N , I CNMI.M* T.l l|. MVCNLMIMKTCI IIPMimi Ml MLTCJKMIMfi iHCNnraiflM in Tmv fMic I tet-MM! IIL INUT CMtl NC> *|IlM INC IN fftiCNMMf MILL (WINN C|*IMt fill I CM. .HiM-IM, INI, CNtMtl,! * ,|r K inn III IffCl > INC* MlfllMI Ml ACCNMIC MMLVHI M Kll IN 111 MIX IfXU ill IIXIIM Mim.iitiii.|i, inv 19- FW- IFN CNfN / t|M - Ml IW flTt MMi Mfl Xtll HIM AN| Wl CNM emit eMtA CMfM CMN CNN ( CM R R Mil It MCI <NV FIN* 11T'' m I MCI IMF IttFO ||ff X XX X VH r| XX kx XX X n -j HONS 215331 mi*in or ftcrctcNtca roo o HfwMer CNITFI*,* 0..ft M CNUttMUl 01IINWWMI *M* *IMNN-n-*IMhNi MTCCIIO* M* flOMTlIATION in civcfllCM ENIIMIT NO IMl* fOONOlfOO 00* tM 1M IMCIMMriM Of MM MLLIMIM OCIffNCC 0100C1*1CO.00CNoooo,*NrMii*fotfifHMO lira. if,* occofortoiML cufoooof 10 CMLMIMTI* 0I*lfIN* OK* 01* fOOOM.O fCMMCHM, ONOOITICO *00 footic*iion to mm ooooo ocickc.om 0*00*.NICOIOM CNONOMT,* ( , NO 0 NOT21MOO M- CMMIOIK * ICTO Of m TMONOt rooooiioo of MLOOtMlt* 000*0IC CM *00*0* INCIN fOl TONLOOINOtCO Ol- BCNM-f-OIONIM fMT If. mi cm. ocmnoiioo mo m*tmtt*m Of OlMMOfONONO MO OTOCNM-f-OIONlNt. TOMICOL (NVIOON CM* .OiIMMMI, CNOOONOV.t *VT *1 OTKTM*IO Of CMLOOINOIfO OIQMT*fMlM Of MO10LT*|0 Of CKOOIMTfO OlMMVl CTMO* IN NCCTOM OOLOTTON CMMOfMM.Oi 121-112.1*22 CNOOOMT.* C .* M.IC.MO * NHT2IMC0 MCNMIM* 10 TM IMONOl MMOltM or CNLOOIMTIO OOMOOMO INCLMIM fOLVCMOOlNOTCO OlNtHfO~f-Of*MTM. IMl CMOilNOfCO oionim *oo morn COMOONO*. INMCT ON TM tOVfOOMCNf 0 M0f2IMCl.fr *L .MO 2COMMON focao.fLMOfon# n v.. itot ra.m-iai 10- fut- TN CI*N 0/T ftMP Mr CNV nr* MM. net* MM Ml* *CC* t*v l*M Til/ fC*a HIM Ml Ml CM* CMN CMN Cue* CM* CM* CM* CM* r*N 10* IMC H INTO III# If I MOHS 215332 i tt Knimui in mi MriHitrf ciimim tfiuriM ir rm >imiimw ra r*rc mci wmiM mwini MW rwt Trf MKH MLlANi FCN.M. INI (WMM.K * *fl Mr MmVfll M IffCMitCM Ml rMMTtl NMCfl M-ltCftMMUWV NCffTtfi || Ml Tut MtMWt m MDUMttl ItNRIt* T-iriNTM III MMIMlII IIMIlM tm MLlTfft CWUMI1, MMI *N TM fNVfHMNtNT. t.MmiMftrtt trim--l MIllJiWMI.* * .MM. M flt'tlJ. CM*.J 1 MM MMICH MNCII M MVCftMIMm UNtNAl <*>. CWMN MUM NIMf..lM|lTNt* in riM hHi.iin. CMLMt.1 ,M| t. IMM IMMvet NfMi $m MiratMtiMro tmmn MTtMlMlIM ra CMMI MTtTCti tiwrHi.Mr.mi--. ,,uiuim. lMr tlYlt.l MITMKTTM T NtlClftNl- MTKI irWMIl. Mi MMIIUMI. itTR MMA tmUMMRMl IMTtM intnia. mumim. vimmi*. RMHIMI IM1CM M- fMN MIMMWII tfMIMTIM.M- LIMTMM. MJI-fl. CMMV.I 1. CBMtttM* nm MMMin - TNf tMTtCMNICNL NITMCTIM M CMtMMtMIM Mi IIMMAL M Mcmtmmmtim tr mi it rm m. MMCIMI net ITM tM. r*- (Mi tan #/i i( mwt * arc mat icii him am act cum cm CMC* emu ca*N cm* cm* cm* - NR NR * * R R m M t N mm aco (hv iim rm i*a TMC* .aat iar mi N g HONS 215333 or fircoiNCc* ri rco an* ru- TN COM 0/1 fCOO 0001 CUV Jtffi ONOi OCT* MM MHH MC* (NV fM* T# T / rctt min mm HCl CON* CHCM COftO COfO COftN COCO Ml COCO TON ft* IOC* 1001 INTO til* ctootv t ,mm # i mm rooiocotm- COL MOCOOffOO Of CMiOOIIMltO 01- ooino CNfooooMttf, tiitr-nt.itrt ctoot,o 0 .OMO i mkmm fnoio- ovtoorotirioo or ckooinotco oitoftonl OMO OtOfOJO-FMftOMO MLL 8N- VTllON 00100 lOMfCOL ,10*110-111.IttS CtOOtV 0 ft ,1 M MO1LOMC0 ONO 0 0 MMO fNVIOOMMfNIOL MNttOllOH HO occoooonoo or oioforootomot am 01otonruooMO cmvioom.ncolto rctortc . ixMOIOtNlOL 10008 cm k o eufoooooo crrtcTO or cto.ooA^NtlOtWt INi CM OOIMOIIO 0COMIIM OMO OCiOffO C0MMNM, IHTOCI ON IMf CHVIOOOMOOI 0 001*10680,81 OL J00 rrofioooH rrtoo,no*roooo v ,toot M 920-91*. IF 0 OMOLVIICOL CMOFOTOti WVICN LfCfOOf 10 m OOS08NTCO 01 INC 900 lOirOMTIOMOl ftVNMOfM OK CMLOtlHOTCO OlOtttOft OMO LU M>*00001, Q0L204M0, OOP 1010.06 TQ08P 100* CtiNMCfT o o CMVtOOMOftOYNL CMOOIMOito ofonino rooo eooniioi - toi itocf CNCNioTOfco or riot mrooiociio. IN. C.OtlMOI80 0100100 OMO tl|0 COMMHUM, IOMC1 ON * n**l000080f 0 WIJIMfO.II OL ,800 rfOOOMOH rtcio.ciiiorooo n t ,100* or iti-m COOOH8FI O 0 .OMO 0 N OfCOL 0818001- ooiIon or cnlooiomco oiovozo-r-oio- KIN OMO OlOOOtOrOOOMO It *001000 M* TCOIALO CNVtMM 0804.10 ttOOMt .CMrtOlHtOlM I0OU8 ft Il-H.ion MONS 215334 at*i* j# irrrifHcn ^ kh OCrtHHCC to- rui- oru cmcm o/t icnr mot Cnv ore mmol ncio mm *mih occo env tono ror/ rco mo* on* nc coo* entn coon corn coco cmh cmcm cm* tm tok inco inrr into *i*r curie * * octaverr crnciftocv or i. J t 1-ltlMMLetHlMllll-MlMtM MON MlT*Vf C*00601 MM* OTMI OftOTlCV- LOTH OHM Cftlft OCTO IfMMUMl k OCCftO.O N t|. i OlkMOMM.MO ft 0 OftlrMCvf iiooHmio or .i rj-TcriocmooootoionrvoMi in to roe* oru, oft MO MHKCVO INI CNLOOIHOTCO 01MIIM HO OCLOtCO fiMNINM.tMMT M TNt MrlOONNtMl, 0 WfllMIMT M. (II HIMHM mM. CUUMOO, 0 > f 1002 N 907-11 H II * or* oofiioo. r c orofoiM. mmuho n mo oioroioi ooocnofo roo ooofet COMTolMlMO 01*0100 OHO 0000*0 10 Oi MtMftTCO T TOC >00 lOTCOMITIONOt OVHTOOltM 00 COLOOIOOtCO OfOfttOO MM OfLftICO COMOWM. MLIlVII.MlIOtl, OCfOOCO (too. ft ft M 01 OOMOlCO ft ,C VIVIMO MO 0 2MH' hi mo iftootooici rootmo in fttocoOIMS IJ.M-TCM COVlOOMMtNTlM. COMTftlOftllOO 01 or0000 104 CtHOtlMtftO OlUINI no OfLftttO COOPOOnfO, IftTftC? OM for (MrlHMtNT, 0 MTIIIOIO.CT PL .000 riMMOi ftOtOO.OLNOrOM. ft V . 1*04 o* 47-04 ft * *i oomoico 0 .0 vivimm.omo c fum- Mi cuvioonocmiol rmioioftci or t,j, 7.0-TC0* Of OfvtM lft> CM.001OftTO* IIHIil M m*ni CMNtMM, IflNCT Oft IM |NV|0#MCM| 0 WIIIMII IT M. fit MMMM V I *04 PP ! -l I J ft Jl HONS 21*335 Mkt* I h OF ItflllMCII FM PCM 10- FIN- ITU mil /* 1(W HI#? CHV NFC MMtf. util MM MIN MCI CV IONS f|f, mi mum m KCi cm cmn tm* cCn cnch cncn cmn cncn tm tom imco imi ihm nt ftlMMK.C i FNf* m IIMMMTlN'a CLCI TMHI if III (MCCtfftltt rnt timo. mm i . imi X mMTttN.P C .11 H ilMlNf. ML.t* *itiMni iimh-fmiim CMircN. MMIItil. CM-4II/MI-II1, rM' Til it fWIMMNfMTftL NMOi l <> IMV, CINCIMMTI. JIMC tMI- * MM MtllTl.l 4 PtMT IHIMIMSi IWl- me in KHficnvii nw. m mm. mmivinc tnc mm, mi to uni mmn .ckdim-imi-h. IMI. N t MMFfC.i l ,tT M. MM IN TM MUfftl TNTCti IMMTtlM. IM M CMttM- HtilN ilOTtlMTIM Vt*NV l-n-IM, VCIMI, MilMHCU, M., FCiMMMT IIH. MVNLl.i 4 ,N A MCT.IM 4 i.RMftCMfi. LmmrMv tM**ct*iMTiM m TM IMRML MCMPM1TIM M MN> Mi NMT10 III Ltm IlMMil M MIMMM IMTCSl MMfOMfta M TM 4?N MM NMMtl IMHIM I MTIJT KM-UVI-M-m.lMICIIN. fWlHMHTN MKMCN UMflMTfMT. CIMINMNTl,liPi< M. 104*1 U * Mmul.l .m N I.MIf LSMMINT IVNVMIM M NttR-IUMMITM Ml* IMCTIM M MlVCMMfMTM MlFMlTli flU Mlftffi CTRMMI (NHN/-n^ IH.RWICINL CMIMMMNINL mmmcn LIMMIMT.CtMlNMITt.MCraCI Iff! * M I V * R R HONS 215336 Mfiiit at HrcMHcn pm mi mmmtMtt _ (MM,a * mMCvuiTi cimrmiiMTiii IN fit IIMHMW iVNfC NTFICt MIHing aaaaa aaattarc* a? act iuimi, WtLM.ICUl.MCIMK l*Nl, NIMH CiCCftlC INttlTNtt cmmmti am imiai m w mi aa nutcimii* aMTfi iiNtirva in MUNM n aa aaNl m T MIW ItNTta CMT m aaacata Mt rav mrticr aa cmmmin ciatatT m. ci Ntturii vatra iiMCia in lac mmm am iwinai in nmYCMatIaaaiCN HPicwia mm act*icn laatMit ica an iiN.rnMMf iw IwiHaiacNiai acatTN MMMCta ccarta, miwmitv aa iiiiaacii. caicatc NLiouaiMiia aimam.ii NtaLVN imcfi mm McaantmtiMM ilia ra/MmrMti turrmi act na* aiHMMlTN. MMtrTVCNtcm.KTCHt tin. c. mn an rm pm mmkm . ?* is- w- ith cat. a/i Tin wai ia mt mm. mn mm Min acci in i* 111/ KKU . H hr a an cim ..N i.Tan.Na.MM i,.thiim NCTICNIlMTINN t M. M*T|tlMMM> NIBCll-#-NfMilN MIIMII M MTNL aaaaacca tv cc-m MMa*aatM.a' Ma.itn. tCNC.a vac aataurt acta aw cw in MINI MM a NNNNINLf NIMtfN NMM.cn - MVINN INi NICNINi TNNICMMIcat aM eacNiCM. mpccti a.cauaNIM.CMNIMI.IM I.MUIaCM cacctm aaaMicaiiNM.MBia tiM.iara. aa aai-iti a ai a HONS 21S337 iiatr hi or iiPiHMii rot ottoPNC# 10Ptll Slot to chin o/t tcnr root iw ore mm mtc NON MIM ftCCO ENV TIN! Ill/ DC* CMM CNtN COCO CNtN COCO CMfN Ml CHIN HMCCl, tNOMI, TtiNiMtT OHO CON- tool fFO-*0St-0i-ttt, tftOMYOIOt N- vtOONNINTNl OfOfOOCN LOO--OTOftV.CtO- rorotno n r .1.0 Tfcoooo.oHO rc MMI OtONINO --o-oo/-o~Yr. IN- OOfVtfOt CNVIOOMNCNTM. OtOCOOCN Ml* OOrOOY.CIOCfNNOJf .tfOVCNHCO tOOt FMII<lfN *1* H mkHPHi frill IV* lcooc or icon or ocvcoo. holy in* CNLOOINOICO CYOOtM NO OCLOm CON. A, NNMI, INTOCT ON TNt COVfOONOlNY 0 o WTYINSCt.Cr 01 .coo >--00000 --0. tutortoo, h.v tiooi. or ooo-ooy MHUMM ocrcootoolloo or rtLV* CNLOONOtMOfO-r^OlOOtNO OHO tOLV* COLONNOYOffOtO-r--OHO 10 COHOMCtOL Cl 01 INC OY 000 Liooto eiOOOOTO- tootov 4 o-- ic. root ch-- isn-isoo.torr riocoTooc.o inc .a,?,o-fcrNocNLOMOYOCNM-tOOO-OYOOtO MOOLCOiO CCVfCN YOi CNLOOINOlCO OHCNOVY OCYOt OHO IMIO OYOOINO C HOOil.CO fCOL, ONU tC|0CIN0lO>.ft?t0-U. Ifro. riMirooc.o .ct 01. ottoootootioo or MtvcNLOoooioffNM-r*ofoiNO oho ocLOTOO CONTONNOt IN CONOttClOL CiOO- rMoco-oni.0.in4 noooc orr mm. cncn , 1 IN ti ** * n R ii > HONS 215338 Nof#n Of lirfMHCCl POO PCI* HfWCI PfffUMMTMOi . 4, TMO MMT||inTl N TCON MOTION OP TCM II to MPPIOMt MOO MCcriMKVOIC HCtHMO I0 ftlMMi IOMICOLlCICM. 400 CNCNlCOt OlPtClR P COtTMCMt,! COVOU.MM. HI 0 UUMH MCCIMM riOHO.MCH TMP, IIP!- PP.41-40 0000,0 OTHOtlf If fdfCMIllMni OIOCMIMPI--HQ PH MINtlTITtM, IHIMHITT OP WU.MMIJMI MO- rut< oti cm* o/i Trno pmoi tuv ore mm. ocro MUM ONTO CC | Mt l*| I * T / ico mn* mo Ml CMM CHCM CHCH CMCH CUM CNfN CNCi CMOK TOM fOX IOCO OPT IMfO MSP 0O0* 0 ,ff 01 OiMtHi0tO HP RMMfHOIIO OIPHtim CTHII1 Ml 0toe010PMWt - O OTICMOIOH P tlfttttC tOOHOO'O OVOILOOlC CM--PIOMJIfilO- 100,1 Ml OOOO, 0., fT 01 IfPIMHI IP NflNfX tNLMMoTCO OIMHMPl MMII.li4IMM.ftf9 OOOTfIMI.fl .*T HI IMMHMHPPIIOOOHT OCTtVtlT IP TCM 10 H1CC Hi CNLMtMtCO MMIM OMl HlLOTCO CONfMM. INPOCI ON TIM CHVIMNNINT MffltNtffl.Ct 01 .CM. PCtOO--N PMOft.UMPOlO.N Vk * Iff! PP *01-4H OOVCV.C. COMtool UHC PCO'O O NW IPO 4 10 OG0CCI TO OfHIT PCI CMTOHtNOTIIH or MvMUMc it hi Pittortci,NOH PiONl N010M MltC MM , >!!, MT IJ.lMI X H HONS 215339 Mtau ar Hriitwii raa ac>i an- mw- iti cm* a/t T*ar nit fw *ra mm ana mm mih acca cv rum uts rce# him mi net cm cm* cm cm cm* cm* cm* cnii fan Tin me* iwt iwi ait# iw,( i ji enMfM ft a.Mrr ni pt* mi iNfiwfiMivft HCTaaa faa aertettan ar NLvmaaiMfaa aimttna aw ranwitvca ta ata aaaL.tM* nift-tftfta, aan fticarav.j r ,mi a a racaaci a pm* linimmv acraai aa tm iwiMTiaN ar m is vim amt rat tm acrtcTiaa ar *aiania-iu(" aeifvnv aaiaa *faaeva ar aaaf rata tm ataaaaaiaa av*Tc arrict aaiiatat Mi vat atata aerv.ar Mat*a,ataaOT,iaaa *,p * acaaton aatfc aai m mmr TBimaaravaai-m raaaaaivf aaaauama aartatM#,**actTaara ,r.ii-ia*ia?*- ftaLftfttfta, j.a.ftf at a,),?,-tct*catataatataiaraaaa a caamactauv arauaaLf mi PM MrMWlMvti BiPNfvn taanta tacarruica aa tm aaawca ar Mftatic ctracaaaM r-ftra ana aan avaaacaaaaa araaaanaat ra tm aaf. aaaa rM,>ima.JiM*M4 . ia, aaccncc.ft p,,aa a Paiaaa aacLtaa Mrfaac ar ft,ftr.a-Tc.Taaa*amafatiitar-araaia in carai/fti a*a aaast* aicti lac ar im Mrarrc cvraaai. aaetafaa pHKiH 4 atat mu tft4iMi4-aatik i*?t. uuaic.v p taa a pum TCaa* a MLietai raaac raa uwcafiaaTiaa tm faaacfiaa ar aan araaacaaaaa avaaaanaat iai aiaaiai ramcatacicat aaa cMaicat acrtcia i eafiaatai.a cmuaaa.aaa a aatit. aatctaiai ran rCoTiaaa.Ma taaiMtri, rr Hft-ut NONS 219340 N*l*l or MFfOfJKff m li 010- ra- IN CHfN 0/1 TOO FOOT IW *T6 MNM. Mil MM Ml 10 MCO fftV TON* Tit/ ret* HIM* Ml* MC CNM CM* CM* CMtN CM* CM* CM* CM* too to* inet I MM T I MO Oltf OH 10r J 0 .MM f HUM CMMUt ID 0*1 MCOATte COLL HMOllMCO 0001no M r 0-tl TMCttM<*rNM>r->MN ri iMtomcot too to* marii tomcoki COL MM CM*tC*L OOOCCfO 9 COTTO* MO1.0. CMMLLMO. MM 0 MiilJM OffClMOM NOfClHflM.tCI miJO'O N OMV.O 9 .KT OL imMIIO 00 OftttOIC 00LWMLOOtMOTtO 0100MO-F 10*100 J.000 COCO .4hl4>Mlt, N 0*000.0 L .*T *L MTNOO VALtOOflOO u rot TM OOrCOMIMfIM Of nTOOcmooo- OlOCMOOtOOt* OT tM LOO OOOfO ft* TO ILL 10* LCVCL OMM CM*..l>tM- I0M 1001 0 OOttf.C.L MM C.ONLOOf. WT1MH *00 OfrCOHIMIMO TM MLVCMOOlMOm OlfMHVL fOtOOIOM TOM tMCIMfMMOM MM COfOCITO* MM MMMI flLLlM FL0OT0 MMOVf^-MI, NIOMOT MOSOOCM IMQTIfQtf, 0*0000 CtTV.M . OOMMMO lilt 1 N0LT00.N t .MM 0 0 MMMO 0 *0ML OVOTtO TO OfMV TM MOMTI0M MM I0L00ICM OOfttLOOILITY Of FCO to OTOOOMtLO till OOMkflC TOOICOLOOV MM 0*2*00 OMLOOTtOO f.L MO MM 4 L.OOOfLIM.tM OOfO OT* 0*4. MV!CM tOCIOTT TOO TfOTIHO MM MTfOlOLO.MN TOOIJltV. ff IP0-I00. * * 1* Ik MOHS 215341 mtiii or irriKHtn rii ni in- rw- in cun i/i m# Pto? *tv *rc nw urn mm mmh tv rM rai/ Kll KINt W| MCa CMO CM(M CNft CM* CNffN CNCN ONt CllfM ran ** iMca ton n* tit' NMtsiNti a.ar at aiatOticM mtivi?T Of TICMMCM MNiN Ittl MNMl ra atocioo im taioosaitaiw in acaiowa rca ia muca caciiicia. toaicotocv.aiiiawu* toi. mooif** a l > mm a o ootato no- mm Nin-HMUT|I M-OHMTMMMV* mnmMia opicioomiiot <oe*ta i op mml'i'Oio rot ihc tottcnci op a,t,?.a- Tci**etiooooiottitp-oroti* < ieo#> in fttvioo*ni(N?ai Mptca mi aioaiti IO*fCOlOOlC. to CtCHICM. OOPCtia P COTTOOftl.O CMM.UMO.MM C OOLll. toe OPictM PONLicoftoNo.m ran. lara pp it-ii HMiiiia.a l .mm t t Ltora. oootrna* f|T OCTtMIMTlON OP M.M*TMI|- CNLOBoorofNft-i*ioort ttaiowo ov ooa cMootaiofopni/nooa aPiCTaomfot iti ctcoaitwtica oioiriNt mm ooloiio COtPOMMi. IHPOCT M fM inoiiommiii 0 ttrtlt0t0.il M .,IM PCMOMflt POtaa.HtftpftOO.t.T ,iot pp m-m taotcaa.o l..it m mmpli pocpono* MOt MM Ml CMtfTOPiOPN* toll OPCCfOOKCTt? OCTCOOIMllOt OP t.I.P. o~TiioocmoMorotNio-P~oioart ohm CNit .OilIltft-I00lfl0 Mn.r i .if at. octiooi oiotooi* cat erne ra op root rt LtMtoioot MM MM a imriOOH NM.TR PfOOPCC ,lk* ptoititiM fcatt iiiai-iM.tm H m * x % " % * n * x HONS ,215342 Mlr* i h r itrftfMCf* fM Kl* ili- FM- ifH cm* P/1 UNF FHfiF iNV MFC MM, RCTi MM MIN MtCi |My I RNt TIT/ Kli fflM W| MC CM* CNCN CMCM CMC* [NM CMtM CMfN CMN Til Till |N| j*il iHfi PUP Mil ,f1 ML MTIMIlMTtM M FHVCNLlllMlTU IIHNM-F-ltMilM IN HHHItk MMICt NCCNflVt CNfNICH IMIliriM MMII MltTMinr IINM CN(N ,|li!474-l4Flfl9M NMMiN.i IIFNMVI NtTMKlM tV MT LrrlrVcfcl iNi IiCf IHinMcImli H. IUiwNiLiiITCinni,t m MLVtNTi MW Ti ftMfl .iliti- HMMt MV * INC CNfMICftL KTtN(ilftMM i Pt mm PtPmtu item FftCil. mifa Ml YMf.fttt If tll-lM, MM I N INMIIM Fi TCWi IN* MktN itur ill tMLNlMTCl IIMIM M HUTU (MFKMH. IMM I M TM CNV1MMCNT I WTZIMC* T PL , 4M iiiMMM Mll.nilFU.M .IMI ii MI-191, N*LLIM,C i MIIICfM miLlfV IN It II MKieilllM M MIL TltN LiVfS CMMNTMMftlT mil ki.ms mmI me .lliTIMO.ilfl mllin4c -MM * c limn Mill- CIM RMtLITViMItMlMlfM IT MIL TNfft-LITfl CIIIWTMlWff. iCICNCC - I Mi MS-MS, MtLIIMC L i ifiTMVIMC ilMlNi N IMl MM M9MMM NMTC Mt,llll| M-j.iMi HONS 2LS343 MTtin m huhrcu ra km KM RtW tTN CWH t/T TKW KMT KM NTC MMi MTI MM MIN MCI 1NV flHl TNI/ Ml OMt ClKl CMN CNCN CNCN CHf CNCN tlHl fM IM INCi INN 1 INPt tIM NtMtM. J 1 .1 J MM9MM.MM | | itic m TNMtitnm p attetM 1194. 9911 <NVlMN.eMTM.TMIC.ti iif-it?. tm. K tMUMtTNM.tt *CT at, OMNICM. Mt TMIC91M1CM. CMLINTIM N iMttf* It NNt IVNTMTIC CMttt MtlVtTIVCft IF tt9VMlt*M919NlN. Mf9M,tft<?tt- m.mt. * NIMCNt.K Kt P99MIM Nt NUI* TIM. NttMMM.TttttiMMtNIt MM 'in Nft VNM, liW. MLKN.I V..MM N MNMM. tUMtf tTMC C1CMNP CP MlMl TIMM IT* ratert ft* MfttMOUtaiNt tctittc MMlVtlt. 4.CNMMTMI ,44i9f'4tl< mt I It a* ia MLMTCtr.t Mtt PMMBHTMMtNP tP TCtt N tCLtfft MNNNM. 1N< tIMINi TMlCtttCietl MM CNtNlCtl MNCfl P.CtTlMCNf.t CaVM.1 Mt.MM t 9M.11.Ctt 9KC1HM PMLlCATIMt. NtvvMc.rm. pp.h-*9. MHNMit.9 .IT M. TM MVttt MCIMTi 1t M19M.mtNT.MM CMMMINctt. nn.4 tec9P.Nvt.,t<3tr-*rt,rt't MMCTtU.t .MM N Mtt INI tTIMfCt M NtCMtlM MTNM11M P tCtt VMCt ittttaitM cMMiTltnt. ini cnlcnim- let tlNNIM MM MltTCt CCPN99IMI. tfIPMI M TMC SRVTAMNCNt t N9T11NMt.Kl at pftt. PKMNNM Ptfftt.KlMtftM.N V ,1991 PP tP-tf it M P N MOMS 2153*4 n -i IMIIII lIF HIFIItlCfl FH Mil orrromr* FID- FillFCIa IN< Ml _ CHIN |/t 1CNF NNI INV NFC NNNL Mfl NCI CNM CMN CM! CHIN CNN CHI CNN CNN MM MU NCCI INF (In* til/ TNN III INC* INF I INF! I1IF m*r.d It >c L*K-lt>N MtWl KM U MM OHO FOLtCMLfttlNOTfO ftlftfHfftFHHOItt INI, MKNCM .IF H OMOIVTfCOL HfflNN rot NfMfClK iimm VOL li ftfTtft- 010HINOtlOM or CHfMICOL COOMtlTT-- otrooi m to n-im.i o roots OTOfCHO COHHOMO.OMftfHO OrtiOOMfOO- rM c ihot ion. It H NwvttNoUiO ift at MotMtantCH, OfOOOMttOO OF o?- MO OCTOCMLOOOftTOfNtOFWiOH fOVtOOO MOOLfH WMMC.JO* rtfTMMH. ttoo* liMMrUtn 0 H HWIIIHOft.O iff OLi *0L*CJO.0tlH0Tf0 HM M M OlOfHM-F-OIOHlHO OHO OlOmtOOOiOHti O OlOOMtVIKOL irrtMCH CMMO- OFMKOO.to*to-to.toot. NVfffMffl'O it.OOFK.OHO V It TOO OHOLVfft OF CNLOIMTII (MHMftTlC MVOOft* CMOOHt ft tmooitlOC CMLOOHMtlOH ooottTorm oho itoocioool oooccvt of tm FttCHitot oiottrotioft of itONCim. . NooofHOLfHf r rtmum. otosm**FUOOH. OfOfmOOtOHfN, MO tot. MIT 4. CHVIOOH OHOL.CMKO..itO*tM.ttF. *** K HVWIMtfi.O .0 OOFK.OHO V fltfeft TMf CNCHtiTIV OF FtO'ft COC MfM.CUVtLOHO.OHIO, 1094 i k aii tHfltOCFOOtHCHIOL TOM fOOCf OH FCM KM OHO TMf CHVlOOHHfHl. ITF-OCOFl-1.0 ft ftCFt OF HtOl tH.fOHCOTlOH, OHO HflFOOC.OOOHlHOVOH.ft C .HOOCH IMI. K MM H MONS 215345 m;|(I 9t AIFIAfNCll FAA Kll tffiiitl IUMII.I AI**CCMMMIATIAM M I.J.f , A~fIfAACMLMAAlASNlfl*PAAA*AIAMIM. IN. CNLAAlNAfCi PHfNAHV ACtAI Ml tNttl AIAVINA C MMl.fl ftli ML. < riCK*LM>, JF'MA*t*. ftM HO- fUM- fell INI Ml * AIM CNF! fl/f IIRI fHCt INV MFC MMi Ntr| MC> CHAM CMCM CNffH Cut" CNfffl CMC" CMCN CAIN Jl nuh Mum free* cv ?*<* ri/ TAM m IMC IMI tHft mi WHIM I MM | MMMtlM fMM MM m FALVCtfcAATNAICA MIHtMCHFUHHAA dem mmim * nf mcciknt im caMCITMI eMNlNMN MLlcnMTMHI MMimi <Ki> TirCniaiMTfi im* '* MM HlMItl CMMMMM. IMPACT MM fit iNVtftAMHlMf I MTllMfl.IT M. . CMM HlfiMMl MfM.IUIIMM.I.V., Ill) M IMl-fiT JAMIMAN t .6 AUNAAIAAH, AHA a AMLIMM Ff-MMAIfMMiIrMliMf AMFMHtfMlVeCaWMMifTMTMTMIIAIPIMIIIICNLMAMFNffHAL flMUTIIM- MCI TMTM. IMIHN .IlifM-lfMIII- JAMtt.t C MC! MMVINfV Tut AMLK AT fldM IM TNf 1AMICTTV M TCfM IN* CMLMMIMATSM AfANlNA ASM AfLATIA CMM* MUHAt, IMPACT AM TNf ANVltaMNfNf, A NVmHCiA.lt AL 'IAI PfAAAMAM IMII.IUNFMIM V .t*A* AM ItMlI KM * IT X X llMWT.f C ,1 A tNLNtt AHA C ML' ItMCIAN HAIliriNCI AHA MTMAUM If CNLAAOAf A*|Mt |M WUl IMV IMAM AC I ftCNNAL .ItlKMAII.im > A RtAANCt M Jl Ai iMVlAAHHCNIAi lAMIf ICAMCI AP CM1AMAM | AMI VA IM* CNLAAAAIAMlM * AMfClN AHA FAII t N AiAlA.AA AAV CMCM MM ,lt4A*-lM, I API I I * HONS 215346 uf iEf(MHCCi t6t Mm MttMMtC $10- *<- KH CNfR %/J fCHt OMi IMV f*rc ntta MM MM|H CC (Hv IIHI It!/ Pdl Hi Ml Mt Wl CMM CWI CNtft CNf* CNf* Oft N CNf CMM TH ?$ I MCI mat |iM4 DM RIMHIV r C .It ML If rfe*CML0Mltf- IW(IRIH IM INC lWflW|ltt4|Matia. Ptl. lM> HCmiMItMItM ItMIMN Nf*t;w PIIIMC HMM fli i*z-ttr it?> MM II I* L N .ft Men ac WI(CIMl rM ltltlNlM lew MM >CW Mwn.II Htmtn IM 1*4IN IM1IMW. WH IM m IW MIIICM CWMCM. MIIITT. IWHI CIK.IIIMMI.MMMMI INI. Nf**,r a <aoa 4 a mmiiCk nltcnliMaiwim^-aiMMiai HtitMtin trricia mm mt mtiMMi itiMu. mi in mama aara im* cmanaaraann item mm fait t n min.II atv CMM at# jiaifa^MjtPi MM a R tttmt.ii 4 cmntmti m cNLMmafft iNitt-p'iUNtm ami actant cm* mwi im imc mcMatw tmtt>ri * CNCMUNlH.MtMI'Hi. t*ai mm MTNUMf m a iCTMCNLtMtl* MNIMIMMI NtMMtK IMINM. CMCM iHVHM>NtaclN.Ui^7*Mr fCff M M H Ml ttiMt.M.t .a n,aam,aM m a Mitt CMKIMMCMiala | IHMV m CMLMMfeat* ft itifiimiMfM mm matt* cncmi* can awnn MtaiiM ptwtc .(natalMurat laanv a<t*i-i?a,it7) Kitaartn.a a ,n n acri--rac( m icm in aMvtaaamcaiai mvik raaa act aitHia %vtm*9tm.*mm*9 tirt, ataaauat.acaitAiata imi II K tt M * 62* J MOMS 215347 mii nt m acrcKMii tm pci# ii* rvi* mu cum i/t tin Kll RIM Mi KCa CM CNCH CM' nvtim.j c .mm * auM *,?.ft*H r**CNLMftlllffftJ*P-lflll1tfi IMUtlf* II VIM MM II V|TM IN* IMUCMMf M MiMKMlII WIMfUWWt MRtl mm ecu.micm. errten, a 1.1 cmmi MM N UMlTMJH* MMMM Mill. Ml VM.IMI. M.IIMfll. RHUIMH.J C .MM N.MtMM S.l.?l-1f rftftCNLM*fMN*ft-**ftlfttt|H. rilUNM Tl MMMIMlII IMtCITI II TMITV Mil curwi cm rmi, mitn.im. MMMM .MiirMN.tlll, RNV1M*, J C MM R.MLMM, CMftllftlt*1'Ml |F NWM TtMTM CfU. itM M Mllatll |T ,t, M*HfalMMMl* >l*l*||Ml|lli Mi II V1TM RNH M TMICMV HILIIilMl.iMf. ITM .MM I MM I MCI INCH MM m M.VCN.MIMTD IIMMI KMN MirvfR NMI CNVtMN.ICl.TaCIH NftL.. I|i ItM-ltM, IITft MTC.I N *. .11 M. CRTN CtfM IMI* CICNCtCI m MLTCNLMI till IIMMSI* P-*tlM|M MM MLVCNtl fNM fLT MR n<Mi~MS<iMr RIM. 1 C* Ml IWIM, NKLIINMM, MM ffttftlMNT, INI CMMSIPMS ftftftccciiMii mumm. untmumi mi m tcmlm mmiii nimkvu < nvtMia 1 fn, cMfCMft. ILL tm 11 >. Cftftft*MI/ft*TlIM.MMI lire M lift-II) ttrt me mmt Mil mm min mci env ta* Tai/ cmi cm* cmh cmm Tm hr twi mr nwi im HONS 215348 mirtix ee urcHNCii rm nv MvrKMrr ........... .. aii- rm- Kta VMM Mtt am cmcn A/r reap ^mi snv hpc mw. nera MCI CMM CNRN CNRN CNfR CNR* LNRR CMfN CMN .. .. mm miir ncca knv tuns rtr/ l T IHCt IHI INF* HF IttKMl TMC IIMtHI* TMlC MM T1LI rtMMlCMM CMVfHMUT.tli 'll! (Ml KUiM.t V MMMIMM MlTMM AUN* cnrnicri BtaiMia iMntnii m min tiasua fmm me cwiHMiMt fiat WNCVI, !<)! i V VV 1R 'R 1 1 viM*raM.a .it *mm4 a m)mh- mm cmmi tv net an taatMiiMirta aim NurciuaiMiM iiiwni. hum NCdftt AtP r r RR RlMatAAMC,N ,V MWM.MM 4 N*A*TMM a*tarn me arccni finaincc ceNcnm- me maw. m cairmwci laicm- meat aariANNi ceereaeme am aeit* cmaaMMtae aipncnvlb immimi ian, >cnicam.inmaia im-mix-m-im. MMI IfM. M It-tl RlMMl .N. .VMM 1 MMH NCtCMMNAtlAN m MiteaiaamaTt* aityyar--av IMMM ACtaiKA IN MTtflll VITN TvaiM cmmmmm, ?>*!-*?*. ram khmnMiI. ,er ai. leveia m pait* cMMmataa iimma in aiaaa at MMI-rfl OMIAAAN NNK IMTNm MAC MAN ACCNMTfANNllT UHW VA NCAA. ANU.eNVIAAN CANINN fANICAl,,lli 4MH4I, im MraMMM, a decamp nptcv apiua at IANIC MMtTmiCia 4 MNlKA PNLINT R MM r V R R R R H moms 215349 (Mill* or fllFOKMCti rot Kla IlfttlHCt kMMMII i L ,* f 4 NfflfNlCR M~ TIHIMTIM Off ttfON- NCNft*, NffMOIM OCtMCMMOOIKNIO-N-OIMlN ItOMil IN Tltrlmilt iDNitl *1 MNTft N|N TlULtON tCVtti MM. CNffN ,Ui ti4S-ftt94,Iftt UMN*O0RI,L L ,MMO r 4NffffNTCR NT* COO CM ON |IMI (Ml NlCCIWf NM OVNiNCIfi or MltNffO CMNNMMTft OlBCN20-N-0l0NJNff FINN IIKM.tM^llfONCHLMMiM*20~0-OINlN MMi. CNffN ,f4'4fr**M. IMl it- firt- tru cmh ft/t tent mot imv urc mmi nett MM Ml IN ICCI |N Tlto| 'i, fC|f Hint M| HCfl CNM CNffN CNffN CNffN CMtR CMH (Ml CMCN ft* TIM INC! I ONI !W| |,t " -I M* LONNNRfCI L L. , UNO V 4 NflVtltl tTN* fNffllfl MM lOCHliriCOTICN IT tiff II NffMCNUMUKRZI-MIMIM IIONtll IT MIIN NtfffMNMICt UMII MM NNCRIO COLUMN 1*1 CMMNM f060NNNV CMfMINMfftf 4 111)-1|, I Ml , LMNMUt.L l .MMI T 4 NfflTlfCI TNt IINNffl*$NffClf 1C OffTffONINOTION Iff fiTtMCNLMMMKMM-N*tlCN|N Of IMF Nil ft ILL JON COMCSNftMltOMO IN* CMLOtlNOIfl OlONlM mm ftfmtct CM MUNtl. INNOff I M ruff | IMIfflNCfft.fff *L iCM NCMMNM NMH. CLNirMliN f .fftf NO 1*14 1 * M IMNNNNORI.L L..f.4 NffITNfCN.MNt I R IlffNL Off ICON MOTION IN NMtT-Nfl* MILLION CONLiNTlAtllMff M I l.M-ri tlOCNLlOOO lOCNZO-N ^OlONlM in MM MM. CHCN it 11413* iaM. I Ml N tOMCfft I MI ML fNffMOL CMNllllt ON CM0*1 NOTH NMCNlLff INiCIMMII' ORlNt - ONlCtN mm NOIK ff H Mill! CO MOV CMC* MO IlfiNJl.llT) RR R RONS 215350 mi*im wt firiMNCCi rot Mai CM MUCK (- rut* am cufR */t tti#1 raor cv arc iwmi. otto mm mu acc env rr*f tat/ Mil MNI Ml NCfl CNNN CNfN CNfN CNfN CNfN CNfN CNfN (NCR 1M tlH (Ml IMT (Nfl Mil immii.m c i r iMtr am a mimi PttiMTiM r *i mum (Mint aim OTNfl CMMNMMM IMMKTI P9M ciMiiMtfi ramaii am anivarivti IMVIIN MCM.TN MHMC .KlHOJNfffOL iflsuc simjih. M R fmcmocm.ii l .am L a imaatf ikoniimiion or raotfl-rtf>tiiiioN CMKlHtlltlMN or tttON- NfN*'. NtP* la-, am ocfftCNiooooiMfNro-p-oiaNJNi in mmm aiu awn tufa ,Ui IMMMI.IHI N NO or rotvcNLOOooioCNOooroNtMO am OOiVCNLONOOIOONfOOINHUM IN IMM IN* CINCOaVOt CNIMI0N0 l. CNLONINNIKO 9I0NIM9 *N0 OCiOTKO COMNONMOii IN- MCT ON PNC CNVIOOMNiNT 0 WTINNII El at til PiOMWOII PNC Ct. Cl NOT000, N V INI PW KMSl I1 R U|Mlfta cv ai f I(L| MNIINfiMM- I ION or ICON |V IN.TM'OI0lKf 00010IIOWO INi 0ION1N4 fONICOtOOtCW. 0N0 CMNicd oortett 9 COIIOWfNf.O camuawi am t eat i j. cot ooftiow PNOiiCftitONi.NCN vooa.topo PP 190*1M N N iltoaCN N niwlim HI * fOOl J.,i lilifMtt.lMl N NR ff iiM^aNL.R ,c iarrf.awo n n Cvtfl mnmniion or roivCNLOornorso oiooiuorWMIl <MHI ON* POlfCHlMINOTit lOCNZB-r-OIORMlO <PC009 PNON INK vroLTfii a# roivciHoorwatf oiPWfNti EfNCM CNtNMPNtH ItAMIMHl MM R HONS 215351 aaixix ar acrcacncff ric KrtKHCC............ L MMIfHMM X. iM'WlltMKMT #T NfMHtH at TM Ml ilMUX iTlMir- IHI LLT 4 X ,KI H. MlVHLaiMr ilMtttfnt. 4 MW HI LUtTKNMMMVI* 4<9 a ,f KII.MI i.lMtIfMCt CNLtMflMm #JMM-**#tMltM Mi Hum CMHiM ta taeriiMTM crnwMra cm-- itaa. imhlc.n .mm t r mutt-- to mtin r emcaniNaxM^ifiirM iv iti Mir--rr-- mm CMLWr--TiMi -- it- r--iMMra c--t r----Mai , a .##*a. INI --,V a VNMlt, NlVMMIIM- tc ii--MNMi mm Hum can* HWH --Tism viTN nr--*. tm CMLMI--rti ifMUM mm CM- MIMM, MM--1 Ml TM IMUXi-XT i MmiNOl.tT at .KM. TlilMMI MCif. timmmrn.n t .tail, m Mi-Ma MMM.V ,1 C--f.MM 4 MiVCMLMIHMirU iJMHM-- TM rri--i m --triura im t-- mm tmia kmitm tmmtm ac*a*itua mi --l avta--H --Mmaac mi* MHTia ai TM act ilMUM IMHJM. tanaaa CMv,tJ*MMtlrMaTMsc im. aia- rui' am cmk i/t tim mat im arc mml hti mm aum acca ixv tax* rar> xx HX XX X X X XX x XX X X MX HOMS 215352 AT* Ilf tW KrftfHCCfl FM Kl* in- rut* itn cncn i/t imr mtt ihv nft *hm kii nwt mii acc* cnv rawt nr/ ret* tiMt mi nta (hm cmn cncm cut" cmn chin cncm chch tm tan met mri into aur MTMNVM.F ,M| N J MNfftl T ict m me iMccumAtiiM ana tictt* tine mcaammm r t.j.r.i'tefMeuMMfiMK^MruiN iwrm ncaith rmrcc..AMiAJNf*rai JIM* MttNfMr* I .tm M CNLMJHAHAN VMM KS UTlFWlIM MA KNCMTIM. li CiMfttCi PM* cmiMit hatimm cMctcnec m tttVCNLft1**re (fHRMlt (HVIIMI1 itri.cMictM.iumaffti im rr ii-m NAT TNI ABC-- i. HHMT MMT IMTMUAlCM. MTA tr I.I.M-fiTMOIliU' MHM-f-llltlN (TCM) IN. AlANfCi iMICUtMfCtL M MllMI MMCTft r cavtamni.a tmtujmt.tm i.mui. m artttowi NKicirraMiti tm, itft tt is*m Mta,t.tKT cnicriimiTiw m riTMcNtuMiitatinMi `Mmm IVTIMCI MTfttCT MINNltl NTMI thc irtitua tr cwmmmmial chcnis- IftT MttI CM CM* ICAt ftCIfTV,MM6f cfTv.NftMuti.MrrtMta m*. CCMMILL ,1 t .tm J.t WM INC IMlClMtNlMT tr TCM INt tltNINi iCRicMtien tm acmeai ttrccia r CtTlMCNI.A CMMiftM.MI a Mlf, trtcmiM rMiiMTitnt.iicN vm.iirt rr iimii. MONS 215353 imTits m MFCtEMCCt fma ka* MrtMtfC IKWAM A TMf Kt iiav fact faam ricirm. kccia.mai, Alt- FAN* AIN MN A/T tCM ANAt CM MFC ANAt NCfA MAM MMN ACC* CNV llt m, KM* MAC AM* MC* CMAA CNfA CMCA CHAN CHAN CHAM CMCN CNCN TAN IAN INC* IH*1 INFA !* AN N N ft ii i r hchiwt,j ,ana s kcnmu atmacimm. akcifictty mm im iimin tCCCFTM fA< CMLAAfNAtIA AIAAJHA mm AClAtCA CAMAAANAA. INMC1 AN VMS l* VIAMHMHNt NVTIIMn.CT Ai ,DI. MMWI N|M.lLMniM,V..IIM A* Sir-Mi. NCRiNNfV. V .(? At AfVCiAAMDH AMA IfUMIUlf AT * MlllMMMNf fM INr CNLNnMTn ATMIMS MA ACVMTAA CAMWM. KMC! AA TM BttvVAAMMCMT MHirWA.ll At .KAA. NAMAM NCIMIMriN.R f jMt H 4F-FA i MAfITA AT S.,?.A*Tf' lAMCMAAAAIAfWB MMM-AfAM|M IN lVJMMIATH. AAMAlfA. JMi tMLAAINATf* FNffMAmr MCIAA AM IMII AlAAtMA c. tMY.II fCAt.AMt carAenMiAl.trr 11-M.lf'A. ACAM IT. 1 9 .1 A NMMTI.HA 1.4 MAMCU. CAAANtC TANJCJTY AT J.a.rrt-TCtiACMAMAIAffUamAA FAN >MIM mo--1 IN t CMANI ANTIN AtAMlAC AAA MtATII CANFAMMAt. IMFACJ AM fNt IHVIAAMMCMl. A.NVTtlMCCrCT A ,Pft KAAMMAN MCM.IUNTAM, N V .IMA AM 4II-AI*. HCNALfY.M * .CT At, KAAfAVtllCA AM t.ir*.A~TI fftACMLAAAAIA*MAA~FAIAMIM fN FAT AM A ANC9AA AANKCt < MACMCA AMAITAI FAAA CAIN IA AACAA 1 N N R R * R M NR * RR FOi HOMS 215354 NNTNlJt F flIMHMIl rn itFumtf .... rat- tit cm /t icm nut in hfc k nett um nit *cc* ch* iim hi/ KM Milt Mt MCI CM* CM* CM* CM* CM* CM* CM* lit ** IHCI IHM l*F tilt NflCtCM .Ci CKITMli <NM/tmcT ftimmuiNtNi rm munkhimfhc acanciwtitfPSNT m a MMtM nmnp m imifi nuimi ra tnc cnnnis* iim cp inf l*w(M danwiTrit at KNMN MKII.fWCH.IMi. Ff UI- Ml. m * a* aNa hilicb i a .i.i.MMii.M t.i.mmh(i tmicitt m tt,Mmiew.w CfCM> M MMTIC IHMimi HUMS--I.NULTN PUMICT,. itffRiNnrn um 111^*191,11^ a aa 7 kills a. MaiMfina hmnn ia Miica fNVfMM.MI TfCNNM .INtlCM' tMf.JMI. aiLLla.a inc pcs inmnslis mmnm act ISSN.. ir< tia-tu. imj ntLLta.t ins MMitiSNi m pnbb LlN SttVIBBN NI. TSCNNM IW a a i*NrNLaNMrmtMiNi r rtm as* CMPMJTINN m H919B ),4.B-TNTCNLN- mpmcnbtc. . tat * NlfCNMR.N a St N TNI IIM MPMTr MLIMUM IINT in INC N'KTM* 9NUIC JNNtlftTlNN-aa NCTMN PM TNB laaaNa-CPSCirrc MicMiNaTim cp I.M.MCM IMNltil T TNf NCC cimiin atnpm inn, *mmm cur. HI SUNNI. MiTfNaa* tMC. a a HONS 215355 MfiM m icrcMMcci tm ki DID- fW' !* CNVM >! If ftp Ml CNV NPC Mi Mli MM MIN iCCi M inn* TIT/ PCifl imt Ml MCI CNM CMC* CNCN CM" CM* CM" CM* CMN IM ffttf INC* 1MI IMFO il*P WI------------------------------------------ ----- ---------------------------------------------------------------- -------------------------------------------- RITCMM t .N RrMCNlt,M t F MICI AlNtflKIIC FHtMM IMIPN* HM Mil IPfCIMMIlT* tm INHCI IFICIFIC KIIIHIMMM P IMCf LfVtLi M M.M-ftM II MU PftfMNCi ftp CMlMlMMti PftfMMTfi it mi nmmiml NfiMM.MittM iMicrv rn mm tPKciiMCiif .iiNNtiPfttii, IM1 * R " ftlvifl M ,Mft I MiNlMli IWimM' IIM M Him tftiitlMTii CMHMi riMl IN IMMRl ifCf ill TMiT CMi- Ei TNt rmm. j.fm m.MC mi. im Hi HR NlVftlN.N .1 IftiftMMli.Mft II MMtt* MtccriM tm KFiMfMftm m mimw imueimiiwfn cmpwmm in VMtHl iiet ilL MINI tut <TMM 1 4 PM ire MC MM. IfilM'lll.llli R i *K * Mifl.fi I ,11 M, cmPIMMCf Llftlli FM IM IMIftPC iltNl IM/Cii OMMtMIMKlC'Mfii MCtftMKlfilC MTtMI- mum if M.i.MiirucHiiMiiifiif* P-tlMlN |l fWlMMITIi MflPtCi PMiCftlEft it IM iCi iIMfi ftVEPMlUR KM|M CMV.MtiWil.MPTMSi INI MMI.M IMKUT IF M.M-TCrMCNIINIINHM-FMMIMIII. till CNLiilMlffi PMlimt NCIH MM TMli ilft IM C ftMEL.fi fCftL BELL tifftCt* NWm.SFi I34*1 ItTft Wh. 4 i .1 ft EMPIftrMft 4 vfti IftMCltT CP t.l.F.i-ICTilMNLftftftftl- CHfi-FNftM - PftCL UlMlT MPWTI INi (MPfftCNtf PftftCffilMi* MflftMi CMaPCPfNCf M PWVCWMIMlfi * PMNVLf IWVTMfl lilt ClfCKI, IlLMMli) IIM PP I7-*| n M V % W MONS 215356 Rafi'x or icriftiucct rot rco* iifcirr* MOM M JT O, CMTMirm tONlCItT or immk mlockooico iimwirWMH IN MINU MOi, Mice. N M' wo Mwifi mm n v ocoo oci.,Jit< mnii.im oto- pm* TCOa RIM M| a OTH cmn |/t ttor root catv me mm noTO mm min *cco cnv ions it?/ NC* CUM COIN COIN CM| CMN CMfN CMN CMN TN If* lCft IMI INTO OtfP a mom, 4 * ,ff Ot rotTOOfOt KfrtCIt or ONTOOOOt tr-00Oil TO *.),?,f*ia~ Tocottooiotiitor*iooio (tcN) n* VINON NfOtTO NRtMC farttlMM10t IIIM ftili-M.IWl a NOOIfO.N ,tr 01. M to itto coomoo-* R _ iioo or rotvtotooloom oroootaruoooo i in rto rttroooiiooo ooo oootoi moot Jg Olt OUtt tOV1000 CONION TONICOt,. X aN NON110r0 .4 NOCOCOON.OMO C MTff. TOiVCOLOOIOOTOO OlWOMfUOOO (TW> FOtONllON rooo ret oiotooo or otoi MO iHVOCN OOti tMVIOOO COOlOO micot N NON|tO,N J NON00000,000 R.MIVMO TMMOTIOO or rOLVCOLOOlOOm OlOtMOriMOM 10 HttORM otoeftoo, NU .toVIooo COOlOO 1001COL ,IOltO~t04 If?? N 00000,0 . 0 .OOOOOtOOOO.OOO c O ONCOI Nflftlit. 1,1. ?, 0-T1100CNI tOMIOtOtOriooi, o coiricM, iwo--ct io tot troioctit or t, t *,4,o'*>,*"-ncoocolo-* ivoiroton. IT toc ollooo otoeftoo OCIO CMN 0COO0 JMIIlOHMIll 0 a OOMLftV.C L ,C l CfOOCl.OOO J 0004 rot tcoloo loom oiootim. intonooc in VOoOOrOOOtO MlOTCNOOCt OTtiOIION* NO INO Ntc NiOOC 4 .4lH?MN,1M| 1 a N a a a HONS 215357 MiaiN ar icrcH( *aa Ki "--met NMaCaiiaacaaa.aafaiaa.mmi i aaaaar* m auTattM.* laaatta oo c-a.r-aicaLaftaiatata**aiaara awa m a wa. s. r. a-1ctaacMLMMaiacw-a- aitvia in caLaaaaiaaiaa - aaTtia mm raic c a atara.tt aaa cava.att ttau-c.iirr Murana.a .a tuaaraaa*.aaa v aaaaaa acriaaiaanaa ar niNraittl aTtta* ait* mt saviaaa eaaraa taaieai . itra. aaaiaaaa.a taraaaca aaa--anaa meraa ar ataaaric CTHta ttmmmm. 11 CNMaerniane aaaaa ar amati^Mi* nth atttvaiivca vaa***** taatai,?fi laa-iaa.iaa aatiaaat (LKCTaicat aawMacraaiaa MiaeiiTiN imv raniita m mtim ar raaaaaga mtnaaraa ctacttatut aatreaLaaiMfo arranm.* treat >t mc la tucraicai tatirmi, aaaatatraa. a c..M fats. aaiiaaat laanmca ar atatra. cmcitattattia aiaataav m i.i.r.i-nruca^aatfMaaa-r-ataata ta aaiaa-acaarca aicc aTa-aa-ai.KTataa.aa , rtMMiT laaa aanaaai atataaca ctMBit ar etaata aaiTtNLaaiMTta aurnia-t-aiaaMMi catreat* raa Tacta crrtcra aa aaa aaa aia taaiaaaatar arTaw.iNi it- raa ca* Vina m i r i aa a a tra cata a/i rear Mat vmv are *a*t atit mm mhn Mca Kv lam m, CMaa CNCM CNCa CNfa CMtH CItR CNffR CMfM ran ih inet tart tart it* a as hn a anv a 0-J HONS 215358 MU ! Of itriifMCBi Fpl PCI* -r--_____________________ MCleAMAL. IKIIMCI CMNCU At CMMM rufCIMIlMTn IMHM'MIMIMi UMIIAMIM TB CWNN1 MMLVTlCAl IBCMIMI ITIMM.IMl. nm,r aaii.aiM ataaAAi. tMMfll IF |,U,MITMCM|.INItllKN-*- HRIR I TCM > T| im, mm mmmit TM11f*f1 MMtfl Uam.FI- cmmirrhi laaiaa mm acLATta cmmmam. wmi m imc cwiiMMin namaiiMi at jm itmn a Ti. imi. F* ni*m Mil.* 1 ,MI F BCATIT IMFUKKC Of iwym iiMcnca m ten tMKirr. IHf ITMfHl HRKILNldl Ml Util- cm. MUCH r UIIMMT.I MMUMI RM I.MiLMM. menM MRU*- TIMM.Ufa TMMIM M 143-IM afAt.i A ,f 9 MAI IT. AIM T.A AAili11(1 ITMIBI AT INC VMICMT IF |J. f. -lTAMIIiMAIMMA-A*AIMMA < ICAAl AMU M.Y ACM ACT ,M*4- Hum ncmai.a a .mm a a aCLaaia mm1IAATC' MMMCTAiK RtMMMM. AftTV IT* MMIUM TA AANAALJAA Oil CALfUBf. 4 aiat.cMta.M3iAMi-4Mt.iaAa NflTAlCK.T .L tMMAMMI.AM* .TAAAam. MaHMsmi fvhlvtk mam- ? ar Tat n TtiAMcataaaaiaaaM-F-iiaxia laaaait aifa iimcmm. caiaittaaitAtiaa via fmtktik mit*i hi fat cataaiAAifi iiaaiaa aim Mtatta caaMMi.iMMi aa im i. viaaaacai a aafiiaMi.ii At taa. mmmm ratil.ataaaMM.a t . taat M 13-44 l* rua* KAi miff mu H i * % ii* cn i/i hr* mat iiw nrs mnh ti am* mid acti inv ini nr/ MC* CHAA CWR (MR ( Cm* (MR (MR (MR Hr th I MCI tuff IF |rl* a VA a 1R HONS 215359 Mma at kfkmncks rM 10* pcii thti TH CM* P/T Til# Ml IHf HFC MMl Nfli HUM MIN *CCt (M Til) t*l/ NCf CHM CNf* CMCM CfH CM* CMR CNfN ClKM TM fan Inca mfi tmt tip NiaiaECK.r 4 ,t l immsie<mi a a ail* avNiiacaia mm iKiTTnctitii m THK IS I(ia*CNLMMIB(N?t-a^IMia a -a -------laawaa at mi ataraaMuci uawia-------------------------------------------------------------------------------------------------------------------------------------------- ciMMiMMMt mm mm mmemtnH' mw mmL.trnn , cijccMatai.ifn. WMiir.a ,at at a natty tut i* atvaiaitic Ftters m km in mmmnli - M aMClla KV|N HIM.TH HtMC , IXMlTMRTli ftMt.St*r-7f. l?l a aiLatan.c a amaua at aiKKMiM- 71 ft# t-aMMMTMtMLaMfCM8M W i mNiafll MMRM-MIMTN. M.l ** THfaia.vNivtaatiT at uaca. aa ca iMti.avcacNiarr a * Niiaaan.c.a .aia l attmm taanitt ataaiaa at TNtuaima in cataaa aataaLa i.twawaiaai .itisti*!]), lata an * aiLaaM.c.a .tf at caaaaaTaaaaaaic tviatacB raa me raaaaiiaa at taiaaa* a tarnat raaa ta&oaa-t-MiciiairrPNtitati 4 tmnnavat* ,wn)M4MaH a aa a aisatT.i c r cental* aat--wT tat a a teat ita-44i/s-w-oi. aaaaacuMtfia MMMa aatTKfv.LTMcaLR, jmlv i#ra in * Mtaiaan. a . MNtitw, mm c Mat fnmtin at chlaaaaiatN2ataaMa av laaMiatiaa at cHLaaiMtia itatan t rafts maaatwttt, a 111 -n. i aa * HONS 215360 IF OEFftEMCfl FM FC|| JUUCJIU. HMIIIM.I .It Ot OYmTHBOI W CMiOIIMIN miHUFUMM AMO CHLOHiMRTtO MfM*~lOMZOBNHM FM IMB COMBOPOnOEmC OIPNCMn BIMBO! MO MTOO-OIPHBMVL-| IMB00 CMCMOOPNfOB .Oilll04). im ore- pm* PCM RlM M| OIIT CMI 0/r TIRP Mil BMW HBC AMOt Nil MC* CM MR cm CMH CMEN CMR (N(l CMCR MOOOTROM, .K MM100OM. AMR OOPPt. OIOOEBO ON IMB BOOnoriOM OB CML0000IOBmobooomo or iooooioiiom oh pvoolv010 OB CMLOOfHOTEO OlPMBMtV OrMBOO CMBMOIPNCOBrOI4l-B4rIOPB o niH,F o o MBorooi clemmp nan* 0000 F00 ICM OBOtOOBO 10 BHVlOOHHfNTOL IMPIII, [Mj OlOMIMt lOHlCOLOBICOL MO CMB0IC04. MPfClO B.COfTOOCOE.O COVOLLOOO.OMO t OOLtE,COO PfCTOHO POOLICOflOMB.MEO IIHJlIO PP IMF Ml* n 0 IfBFE.P 0 I00CB COOTMINMTB In 0 PBLVMOMlMAIKO lEPMfMn BlOO OtfOOOMl ONO 0 OSOOCO PON MM COnPOONOO (R IMPEROMHIRIOL tOHUKO ROLL BOviooM tonion ionicol iiniMn, I0B0 RXX X R OLIB I ,m LMiBMNWfO.MO 0 00TflMM POLtCntOOlNOIBO OIOBHM-Polonioi ono ottorso compommoo im in- CIMBOOIOO EBBLUCMIS lOiCMtOOIOOIBft 01001MB OHO OtLOIBO COHPOMHOO, IM PACT OH TMB OHVIPOHHIHT 0 MflllWIO i ol .boo Ptooonon pmoo.cloomoo. M t .1001 PP BBB04I * XI MIN ACCO EMV T0N0 T*If ion IMCO IAPI IMOO 11 IP RONS 215361 Hit 11 at HFflCNCd FH rc*a mtfwurt ClI- BIN- *IN CHIN t/T IMP PM*T CNV NTfi MML Ml* NUN UNIX KCI EHV IlHI 111/ pci xim Ml* MCI CNM CNCH CHIN (MR CNtft CHIP (MR CNfft TlR TtX INC* tPP? IMP* lltf UI B . P l VIMHLtH.M* MTIIHI* Bl Ml* CNL*MtlMNfMIMMt* MI TMI CMMMMItIt M FLY MM MM Fine |M M MM NMNl*CINi INCIMMTMI III TMff NtlNt*LMM* CIMIHMCIf. 11IIMII. Il/r IX X ItlMiLN * .MM * C ItfNCD PCM IBtVKCHM MMM )*.t**i. III * X X MW MM NKM.IH BflBCTl M MMCLCM (Ml. * raLYCMMlHtin IlMMLf III m CLBCT*TCM. MMMtftV MCM DWltM IMIVIIII.I .MM I LINK*. KHVIIft* MINIM. TMKM |N 9 IMH|*N mKIMl * HUM *N ffHC tfeCMM m MI*MB C*N- fBMl* TN FI*N MUM IflHIII. CMCNMMMM.4tI4I-44l.IM*. X MfMVdH. i .IV M.. Ml |HC UNIT *P MICC If Ml M VC** IN CM CMMUNfM**MV CNtNMMMflC.JM. IMF X PfMCNLL.* * MiVMMEMII* *IPNt~ mil MtWMCMCt MM IIMCICH X X X X XXX PtfTV.rf ..CT M. |H* CMMMIfl*M *V PCM MM PCM NUMI IN BC*IMCMfB M INC WU4VMl MM !!**< TMMIC BINCM PtCWNU* NT INK NC* CIMIIN KVN*MIMM 1MWC ClTT.Nlt**IMI.*ftPTKN*C* INI XX X HONS 215162 Mrim m bifihhcci Ft* t* muuu----------------------------------- FdiLiwr.il ,n m. r*rt m hm id DICNIIIL CdLTVdC dd* Id Mil dMK LIMMIMV CddlTliM FdCKdTft *f TtTd UN ItPIIMM.IMICK.WIflfl' Idd Wfllllli im FiFfi.d d ,4 t tdtt.ddt P.4 tfudfut. tittCTTBd mm limit mnitmid m t). *, t- T(TMM.mINin-l-*ldl Id Id TdC ddf Id' CdLtdttltdJdi - Hicid mm Fdtc t.d .ti*,rt *H.am cd .imiMMiri. 4fv ijuifn, i, numni* t tcm mi Ttm.ddM.ld dd tJLCCd dd ttlL. MU BdVlddR.Cddld* tddltd*. ,||i|M| i FilddCK.i | ,fl *i MlflCMIlMlY tr tiNMtt*p*iltdimv iti cumtltdidt - tftltld d**t FMI t l tLdl*. It ddv Cdtd.Mt ISIiM*f4,lt?l FKCHIddf.F |J,7.|-t(TM(di*M|l` tCdlt-ddtd'tltdld tCCtdftdldtT ltd idi CdLttlNdffft mwtir teitt ddt fdtlt tlddldt, C MMML.Kt (CtL.MLL I ftCCMtUi >,II *1lilt FtCCditdl.F CtdTidttdCY Ft amId* in ddt tctttdtt it mmaeivt ddt dCCIttdlt IdYtLVint TtdIC ddltdld*.*. ft tc Fdtdtdftt II Min Idffft* RdfItMdt YdFMIdd td CdLttfddlKt titdidf ddt mun ctdtttdtt <ddLt* tttt.MKTtfd.tCTttti Ittt II- Fdt- Td cued / ltd! Fd*i iHv mt mmu mi* dUd dd|d *CC (MV IftN* T*T/ N|i Kid* * NC cddd CdfM CMd CMd CdffR Cdfd Cdfd Cdf* Hd t*i lues IFFI IMF* *T*F *M * i km KI aa HONS 215363 MTIII ft# ftiFfftfNCCt Ml Kftl MliailCK raCCNlMl F , IIUM.MI XMMKftl MNUMf NffttfM IFTICTI Fftftft ftCCIftCRfftt fttLCftftf ft# fCTftftCNLftMftlftCBtt-ft-ftTMlit hcm> *r ccvtftft. irftiT mm m t ftCftft ftCI .IlliKMtl.lirt Bift> rw- FCi* *EM Mt ft *111 CM* */r few Fuat chv mfc ml ncrc MCl CM CM* CMBft CMfH CMlIt CMf M CMM (Mil mm amih acca cnv to** mi,fftK TO* rMCF (ftFf IMFO ftt%t ft FftMt 4 .mm 4 ft Ffttffft ft HFIft MTU Fftt ftttiTiM me WTMiuia m t- ITMftxrftCtftftMFEft tv ftttiftiiMftt W9CHllfftft FftftCTIftMt MML ftlftCMCft .lft? tlft-199.rftift II Fftm ftM ft t . ftMft ft c rant ftftCftftftft- nftn ftH cMwfticn> 12fttin m wftii* Mill ftlfttftlft-F-ftlftMfNi 4 ftCCIC. Fftftft cmch .ii.iHMm im M ft mini m ,M c tCHUttm imNicn KUOMflM ftf KN III ftftfi MTH. aitilM-llMin ftfttfttft.M ftMft c ttMLfttTfft INFLIlftMCB m ftftivcutB ftN aftftouftciiift m Himl ftMft iMfBtflMftL MMtlTlftN ft# tCN Fftftft CftftNCr TftMlCftL .[ilTMIMtlt FilftCft M If ftt aitucruftB CLMCIftft* MftM ftF MftMMfttlftH TCftft-Nffftftftt1 f(ft FftfftCMTCft ftt fft iMTCBMftTlftNftL Nfttl* ftMftF QM CHLftftfMftTCft ftlftftlHft ftMft ftt* tfttffft CftMFftVMftft.fttLiMftrftM.FlftCIMTft. CTftftCft Iftftl n ft i au ft pn FftlCfi.it CT ftt ItlOCTtfftl flMCIft*' TlftM ft# MftfMMLlftft TCftft^Mftftftftt ITCt KFKF IfMl1ft. Jft fftftft*4ftft. Itftl ft K HONS 215364 A*rrtf IF IlFCCCMCCl FM FtCl J&U1IML MCI6CC.M . M MIN CMC C KIMITlI MfClM. ftiMtH m MtMCilCH MM I * M(l ICC If ICC# IN me MM MTC M* miifRt #c TMicrrv m rcM-NimLrlfll* II MIMI FIN INiCMLCaTllftIB* I'lMIM* CMC tCLCICC CMNMIMI* FMC1 CM IMf CMVtCCMMMr C tMFUTMCCC. II M .CM MCMMM IMM.IIMHCC, m r ,lc#C. Pf HMD MO- FW* FCIt Mill Mi lit tmn ! riiv FHor iv nrc mmmc Mere HUM Mil HCD CNV IKNI INI/ MC CtM CMR (MR CMfN CMCII CMC" CHIN CMCN T|K III IM iMMl I MFC MM MotMte i ,mm c uMi i.J^r.c-rco TIKW.IIIIINH|l*lMI*lllall Mi CM* 2VMC INCUCltMM Hti CMLCCIMMTffC CMC*mxv MCICC MU fHf IM CIMUlM c MMliL.CC (ML CUU IlfMliauD.fFi UMtCJlFI ff ftltVCM CULM MMTIC lMMn IMMKIIMI m MVL VlCfCM* MM 4MMMVUU MTMITtil CTMMV Cf INC invciiMcmTimv klmtimimip L.iJiCi4-cM.ierr MUM I MM 1 ttMII CHLMCMMfCC eiNNJC'e~eiMMtNiiTCiir incucccc m C * Mil MMLCvvl 1C MCI* CVMTNCIMC MM MtTL HTMCCMMOM rrCMVTLMM. If. C CUM* IF INC tfftUCTMK-eCflVm tClCTIMMIF MCL FWCMCllr.lim-HF, icn MUM * .MM C ILIKC CMMMCIMM M i.u.i* tctcccmlmmm immic-f-cicm a CCTCNT IMMtetC cr MM MVM HVMOmill M| iM J-NCTMYLCMtMMTNMMC MML MMMCHr.llON-MI. ItM HONS 215365 RATHE* Of KftllNCKI FOR SIR- FU* *TN CRCR i/f WAR FRfil I MV RFC Me NCTR hum *mem *ccr CUV It** nr, FCi MINI Mil RC CNR# CM*A CMC* CNfR (MIN CMCR CHEN tMN TRM ft* INCA TAFE INFO tl *Ri AMR .AHA C UHVCH ATARTAR AM IMl MECHANISM RF THHlCITT RF TNI CNLMIMKI AIMRfi-F-RERvfNR (*yh MM MlAt TM TIfSFIC . CMFfR|Nf NVAL Miut stiirtst.iin * AM Mlttlvf IlCWtLOCKI, INC CiMI MCTK* FAA ANR-RAV CMtM AM AM tfCALAIIANS CUT AF TMAMFRT.MLEFRAME A. JAMAH* EARS I FAMEAt A * AtMfSMART AMR CANTRAL RF FRLVCM.RR|H*rct AlFMMVLfl CRCARl. |Mi FfftCVtAIMRA. EITN ANNUAL CRTEARN* NKNIA4. RVRTBM RTRARRIHB. AtllMClAH. TA , MRTCHRCR lA-CA.lSAA AMIftCAM RfFCHtf AttFRRIRlEfRR ARRRCIR1 IAN. ARiINRTAHi.TA AT A-lE K FAKITAA A R .ANA 4 R ALL.CM l.l <9,9 -TCftAtMLAAAtfRMCEm t TRALATERM AMR lACNTIFTCAUAH RF NR 1ARALETO KMtAA- TBR AT HAT LETVA R1CARRRN1* SARI NfTAR SERF 4AiIRF-tAJ.IRAR. * A A4MCL.C CHLARlHAfCR FNCRART MIRA ANA TNCIA ATARtHAi CAMEATRT--AMHRRHT | N i CMLRREHAIIR HAI* ac ear ANA ENCIR AEARIHA C.RANCi.CR 1CAL AMU I KTRCAHRlHI* tF IS-1 F. ESFt MR MV I RAHIL.C CMLRAEMiriA FMlMAMT AC EM AMR TNCEA RTRMEHR. CAHCLNAEAM AMR HCCAHHlMAA I TANA INi CMLAAtMATIA HA* MART ACER* ANA TMfA AtAHEHR C RUA-HIIILJ.KIAEIKCAL Ml (AtRCINtLMl.tFi MR K km I F-48 HONS 215366 F-49 MTIII or IKPlMNCn PM Mil III- PM- Ita eMI l-*T TIM mot IM Ml MM NffTi DM MIR MCI IHV ItNl til Mil KIM MM Kl CRM MR MR MR MR CHIN MR CNCN IM l|H |RC0 IM> INTO |IP MPM.t I 1 r. O-fOVROCIR.OOOOIOCH'" M-r>OlNtN fCOR - iNtlMMIIM IR OlOOlRi lillMNICM. MM MRtCRL MPKCll r COftflMtNf.O mmumi.mm Mul.lll iMtflM lMUCIftH. NCR VOM.IIVI M 0-11. MPH.t CRffRtCM. OOCCOOORRR 0T PM MfRORY RCfOO MM IIMtM 10' CMLO* IIMTKI ORCNONV KtM MM tMtl KIM C IMIl.il IM NU fMIMHHC.N) JPiM-JO, IIP! MRt.C. OfCMlTMlMtrtH IP IMMCTI FMMI MUM INC IMMttlM # MMUM 0# l.M-fllCMMMML EMi OtORIR' TRMCMMtCM. MM MRICM llflcri P.CIVfMCIM.I.MVMLMI. MM 0 RM.U ,CM. CKCtRHR MJKICA- ncm^RiH tmmim. m.ipi-iii. IMM.C IKfNNMTII rtMl Ml- MO. 0 ENHt-tlN.I?44lllP-ll,l IAMB,! NMlMRIBMR VIO MMCTff NCR HV OCR MIRRCRO MO RRt* M|VU||- TOf1,MCO,0WCftfH. lM IMM.C, MOO#I OR INC RNRlVftO 0# OlHCRMMtOH tOICJ MMf. ROT 1MI. ,C MMVC00MY M UNfft MMCORtKO F00 RMITIII M OCOOO MM 0C0T0 tR FLT ROR HIIRIIL COHRNMECRTIOR. ROVCRMO 1001 C r* COM,MM-R ONOCO. IMKriruoTioo or rocvcrlorirotco oimrMCROONS tOCOMk in CMNlSMtOi chlo00MRRL FORUMrilM PiWI-Vtl.lIPl MOMS 215367 F-50 IMTUM BP iKPtKNCCI FN XC* mmmt_______________________ PM t .Ml* C * ULlMi ai H* Piet IN Inf Ml CMIMtMMMK M> tUNIMIlM M II9MITIII III Ptilh tHUMNum j tmm**MM.4trtt4T- mjin BIB* PCM WlNB X BNPXt C . Ml N N MMl eUlNICM. Ml NHIKI MM MMUlftCM. MINBBft. INi wHoccMtc* ftthcmMimiNni, MPNIRBLKIICB* EKMMIMMNC MM cc- LNTffB IMMCM I MlMMMN.il LKVICB/ttMtX MUM* IIHKIIMi mil .MtrtMNl, 1MB 99.4 i -IB XXX IINXPC C ,MM N B MMCB MLIClMtlM* I KB BIBIIIH MM BtMtUOTMMM IN [NCIMfBxrB* CFFLUCNIB. HCBlI NX BNXH.C .MM R NTtttM CtilllCfeC MMtTBIB BP MMBN MMUli fBCNTfXICN1IBM MM MMTinClflM BP NINNLNtMtBB BIBNINB MM BIBCN20PNBMM MflMtCB NT INK B B -IMIIM NHMM.MT IBB} XB BBXXC.C .ft Ni NMiTlIB M XBUXMLBBIMfKB BIBCNMCMMM IB VWIIB BIL BifnS MIBN BCBNLRIIBB BN* CRBBWBIB- x X tfMM.Iitlt'IU, IBPP. BxXPK.C .IT NL MMiTIIB BP MLUKIBBIMNTKB BlMlIB-f-BIMtM MM B|- BCRKBPMMM MfBBNTBB NT RffClNKN PNMUNC MIIBNIP. BtXWMTV MV I Ml XX BNNPt.C ,IT NC PBBMtlBR BP *NtV` CNLMIMNTKB BIMNM-X-BIBNMM <NCMB> MM BtMHIBPMMM <PCBP> BV BNMMNB M NKNtlMC CNLNffBPNffMBfKB CNtM* BPNfBt.tiItB-BBI,IBPB XX c . RenBr* cncr b/i nut pnbt cup $t nmm NM MtIN aCCI CNV THt T*l' NC CHM CNfR CNfN CH(a SUCH CNCII CNfN CNCi rn tax incc tNPT (WO OllP X XX X I X X *X x X HONS 215368 (Mum m icrciCHC( rot pci* myfm bio- ro*- OTM CMCH #/T If NA ANOT fHV mC AMAC AC IB MM Ml fM KCP CUV I BNf Ml I acb *(** AMI M CMAB CNCM CMCA CNCM CMCH CMC A CMCM Cuff* f*X TO* IMCA jurr into Olid MdPff. C .CT AC IWHliriCAIIMI AA PMICHLCPIMIEI BTBCB2AAUAAM < ACBAB > AtlAlNCB IM AAfEUfB TTH VMM cHcMifMiK. 11 >imm . nrt I NX AAAI C ,K1 AC IBCNflAfCBTlB* 3A PAL VCMLMIMATBB BlBB*AAUAAM |H lAVlBBNMMTAC BAMACf MtIMt If11 IM| AbAAC C .11 AC INHIirrCOtIM BA AACVCMLBBIMATCB BffllHf (ACBAB I AH* BCMMMAMAMf t ACBAB > tN MMUM BAA* At If. BCCMAAITBNHL fUABBMf AM VMM AATICNIB AffCBCHfCB ft M IMICMATlBMAI MBIBHAA AM CHLMIMlKB BIAXIB* AM BtLBfCB CMMMft.MlIMtM.Vll* fEMlA.fCVMCB IMI ABAAC C .11 AC ACCMAMBMAC BOABBMAI V# AftVCMl AMI MAlIB B IBMIMB AM B1- MNMAMAM IAt CUNIMTCI AtMtM AM BfLBVff CAAAAAMB. IHAACT AH |Mf MVfBAMMNl B MIBlHMB It AC .Iff niCAMH mcbMlwhmj v mi AA Of-BlJ i N ~ I M M BAAAC C .It AC. AACVCMLAAIMflB BI*M- xM M I MB. IJNHHMM AM BfNCB AACTMMC* IBM AMMTtCB AMMB BMtM tMClMCB- AT|M AM ACB MN 'AMABtAT IXfCM BB MtlAACT * ABCMHTIf MAMC TMf BtVlftM BA CMVIBMMHTBC CMMIBTfV. AMBltCAM CMCMfCAC MC l|1V. XAMMB CltV.HIBBBMT.MATIMMf IMI ABAAC C .If AC AALVCM AtlNATAA ItM* IM < ACBAB >. ATMUIMMAIVB * ACBAt I XN l X AM ft mb aacvmmclim MBMTICB ia- NAB) ABAMCB BB|MC ACB AIMB Ml CflAIR x N u HONS 215369 MltlX tt tCFCKWil FM K(t AEFUIMCE . ... ***Pt.c .a a MMti mm a a hunmi!. IlKlM, rNNffrfUAM*. Ml tTNtt FOLVtMiaCKlMICa HMMlICIi rtMlCTlM UK rUMllM, MM (1MCTIMI *m H t KM Ml .lllil-lMm io- Fitaca* vras mis iM M tia cnr t^t Tina awtr * nc Mitt acrt mm mir acct mv irmc ir^ hc* cum eaca caca cnr cncr cnn caaa^cafa iw im ini imi mao ona M M ttaat.c .a a mu mm a a mhmhi iKinnciriM mm MMtiPicifiw m PKTCIUIIMTKI UMU^P-IJINfM (KIM) MM HMIWMIIM <PtKf> |H i 4 5-r-itfc* nawuTim mm acatJ* CltC MMCt CNSllMM|||KaM9|>4M. lira n ii a UPM C . WnitM.MM a a Nilim tviar ufMUMin pim mcmi act- MMMCI PKHMMI (IMMMCiriMF IW ir a x ErMI, NVMN IMfUMllMM) !VKV IKCIIMB la at mmtfl if TM mi latfaatfiaaat immiim m cm*aiMitt aitKMia mm tuaift etaimn('|.M7m MMitla.tctattt taaa a a tECClMII 6 lOKlCat-KY M TCM MM MutTii cataamMti atfftvaitaaa m IMM EMi (ItltlMlEI aiMMM MM ac* ittKt canaaMMt, irr#ci at tut ih- vitaMMtf a Numacft.fi m tat actatatu aKKfc.fLtttMa.il t . itat aaaa a KNNII k .6 flUMtaiatt MM fllMMHVMII IMlIIIM CMFFKlCaVfl tF tt6miic enctECMt Mama Fta* aivuua NME INI** LIVE* CNtMtltCtaaHYi cvatiMuaii of ntfMtta mm MPucmti tt attflaiMii isviafl fmvcitMfUtica *4**FFEtffl im SMC FCI atBfllltfUfl CNCMtancFf a taj-4tt.iflaa n a HONS 215370 M *|H 0F 0fFE*fCEI ft* Kll -JKUItmi------------------------------------- fl|4* CLMM *44jr 0C0 044I04W Cl TfCNMt ,10.t~l*l 1*7* OIM CMfH <"!610- FU*- K40 I HO I (NV nrc *MAi. nc r* HVN *N|H 4CCI (MV I0MI HI" t0M 1SJI IX. Pcit Mix* Mf HCa CM40 CM* CHfft CHCH L**fW rwtx CHfK CH|| 1401 IMF 0 11P 0*TH 4 0 6 .444 I N 04Ht04HT* fMC F140r 40 CMlIIMTEl HlWllftHWH IX JtfiMH 6HKWIIIMTII - FMlwn *00011 4UU KM1IMM CMIM t0kicm . i*< jm-Jii. rr< MV MITIM L w KV *1 Mn *fNCH* 0M04iTMiHVNMiMtmtimi u(nMKMH) nini n(crttHimu itbn- nmcimim It 0*4*t*it*it tMICOlMt, 'n 23 \ *J-t 14. IM1 (2 ITM J J 4MI J c IllM tNlMIMIM -IHMIMIHIM 400 -HMUBIEMMI IN CH|C*0M Lffft* IIMII 4* It IMG IBM 0tNT0C4t0400Nia0L CMlMIMrlM 0F Mm KIMTIMI 101 CNlMIMfll IIUIMI 400 NLIIM CW0MIH, IMMCt *4 10f IWMMMMIIIt t MUIMNM1 41 III HtdliHUl IHIMLHIMII.II t I Ml 00 IIMH N * M TM(MJilJ0|00 4r1 t 0>. IIW,1H.*T-C0FI0M4MCB414W4` tKMM IIlHi IHIMI IltEMOCft 4MlMCf * NEHMfll 01004* IKK IITIIIM 40 E0VI0400I0t0L CNfXlf- frr MIIICM C*M0IC41 mciiit.kmmi cnTjiHMiMiitimi mi *401, lUCNfllCN IFFICTII KlIBM IKCfUtt 10 Bt NEMRIII 41 TNff Me |0fI0tMf IMMH 0TN040IU0 44 C4L00MM- 1C0 0104140 *44 4CL41C0 004044440 04ll4u4t>*4tV0|0r0Cf04fi 1400 M K HONS 215371 or tirilfMCt foi rto* 10' FM< Tn cmm i/r Tin* hoi Ihv urc M mb to him hhi* *ccr .txv rni it PCIf JMNt *MI HC CHM CHCH CHffH CHCH CHCH CM* (NfH CNfH TlH TOM IKP |MT two ** jr hi ocm * metOku bn- im iMOUCCOt * TOHCVHO(*HCVI V|TV inki in. nxicotoc* or mhumici HrOIMoMCHli NCM.BH MO (tNNKN (MCtfi R I I IMHN HMO M BTHOIOM, (I rtKHHON MCOO Ml TOM r fOOt JOi-Ht * HPf I Cf Hi MITIMCOHHIMOH V MMWMUI HOOfMffC CMNlMM (Hi imhtiricoi ton hho hhhltiii or hili* fHMIt IH HHlKS i M MKIIN.ffO HIM MM ClffHCK.HMM MHMICI,,|t>f r -r. M IMK IK BI Mi. MM M HUM THOMCIM IN) CNtOOIMHfEO OfOMfHO MO ILHTC* COHHOMMOO. |MMT Off fNf li* VlOOMMINf 0 NMf>lMCO*B1 Hi Jll MMHMOH MlH.tmMM.R T <IOft. M MMt MMt'M.B MC TOVCfHOB MO 0|O* OMHICHi CMBCfO or fCOO J*| OtONlHi TMICOiOCTCHi HMO CNfNlCHi Mirren r ciftMii,* cimumi, hno o CHtu iii inctOM rootfcnfioot. mch took,nn or iimii. I X fHMOCIO, 4 RHC OfvtM EMCfMOV BO M rOBMHlB* Of fNf MH lOfCOOHVIOMHL TOTOflM OH CNLOOfHHlCl IIMINB HMO (1HII0 COMMOM.|HitOMHC.HMfOEOr 0C1H0C0 I00 I IHTBMmo HO J OOHMH.HMO J bhcmo rco cooihhimhTioo im oiothiOUT I OH tOMtrOHNCtl UNTlM KI HfHtf*rHir>4J~*0l , lOOt R K < MX X F-54 HONS 215372 F-SS NA1BIH AT iCfCitNCIft FH FCIl mtmtttct it- rut* IFN CMCA i/I TINA AAOI lltV MFC ANAL NCr MUM ANJH MCI (NV liNft TIT/ PC** MTN> An| Ml CUM CNffN CNffN CMC# CNfN CNt* CNCN CHfH TM IflK IHCB IMF! IMFft i I IF iwfHM ,ir n niMMiTaii m ml?* GNLMJN*rift IlNMirMM MM IIMNlI* F^iiftiiiN unmrtCi in nv mn mciCMIIi AT TNf JM TNTCiNATlAMAL FlaiM M CNLAMlNAIKi JANINA MM II- tilli CMNMM.CiilMlS.MIIliC* ACIAMi IMS m mi - ACMCIfi.A CMITAMJMftTFAN A? AN #F * FICC MIL!IIM IMTia ACM, AIMMM, ftl- >HN AMlMI, MM ItMIMLIlil AMf M LlCTftJCAt TMANAFAANfA CMtMlM IN ilMWiMTBN. MV VMt Ti M MCftCNT* Ci AT INC W IMTCMAllBMAi MMMi* AN CMLMIAAVM iKCNJIM AIM MlATKft cmmmii,miicn,mmiiia,cctcmn I Mi MM KNLAirii C.,MM A AftJAM ANIMAL TiKlClVVi AtVfttt LCCTMf 1ft M AIMMNISi CT TM N iNTCtNAllCMML tAAiilNN Ml CMMlNATCft AlMNlMA AIM MLATSi CNAMIM, AALliNAA, ANNlilA, ACTMCi IMS * X HMRTM a ,CI At TAN 1CALMT M CNLMlNAlCi ilMaUA-A-AIAMINA IN CftAMAlftllM MI6IN AIM MK, K N ALA1i,KA A*V CNCN Mi 1W k X CNHBTi4A A ,K1 AL ItolCALMT At CNLMINAlKi iJBCNM^-BIANIM. INYIMN MALTA ACttAtC ,CMAflilNANfAL riMN AiiT-M, itri MiiarU A IM MIBAAJAAFIAN m C, I, r, ft-Ti TAACMLAAAAIANNtft-P-I I ANTA IN AAHAA NILA |N ACftTICJM AAALTTI cal MINAMlMV 4 MANVCY,M ,M a MMK .CH ACi AYNA Ml JIMIMH A I R M HONS 215373 F-S6 i XI _IIUttiU_____________________________________ tUMCTf ,C * ,MI i M.NMMNfl tm mtsmimtim MMIC-P'CICXIN IN C1CLMICML fXriMTi ir ui cwMitMMm mm# mtllMiriV IIXHII IMM fMCTMM Gi>-ll,t*rC tit' rwc- etc tm* i/r mu nii cnv mt mu wti mm ilM MCI CNV f*M* nr/ K*f l|HI Ml NCl GNMM CMM CNffM CMCM CM* CMCII CMC* CNCN VMM ICm incC |MT IHfO our *M MNNNNM,C C ,11 ML CMLCCCLTill am.it* rc cnc ccmtcmimi chlimcmMM CBtlMIC C*M-CM/-*C-M, NC8CM$T-tfNM CMHH1IMMMICMM CLIPM,M 4 ,MLt icrc. V * ftMMM M M ,M M TMMC. MTCICM. M CNCNICM. MMCTJCft CC ClCMlMC IN lt< LCT1CM TC TNCKC IIMMH GNCV1CCL INCKICC CKVKC1M,CfMTCC CMCNCCCL PtrVtICC, NMV KCMCL MMCCCM CP rCMNMl , MMM1MTCH, ,t. * MIMU.R f ,C i i Mflll CIM- IMMtPiMttTJCN CC Kl KM MlNMi M- X *X 1 * M rent IMJHH Ml VUlMLoHtMIMf IMI tMI TM.l * ,M 4 L CUM CVMLMM- riCH cr iNfccfcccMccc rccc mm* CCC1CC cr PCLVCNLMlNCfCC MMMIIC GMMMM IN MM MMMLYfltML MTMMC fM MLYCMLCClNCriC CIMMXCFtflMMC CMC CIOCINS MCCCNTKC CT TMC MCC tTCCIN HrcHHcCcCallCiMm.lfMi MCMC CITT.MimiMr,CtC~ X** R MIC* L M. ,IT ML ICCNTirrCMIlCN MMft MMMI1IM1IM CC MCLVCMLMMICMMM'* CMCMMC IN KNVtCOMNCNTML CMMLCC HI- CCNCCC T CfTN NNMMt NCCI INC,MNCCI* (M MtlfTT IN HMCi tflCIIMNTlV, nihmmcmc. ini * MX HONS 215374 uinr* uriKKii nit HFIHUCI ili- MM- *IN C*C /! ICM MU' IN* NFC MMC NTI MM tkttltt *CC> IMV INI lit/ FCIt IMt MM HCt CHM CMC* CM! CMC* (Mil CM* CUM CHt* TM Til INC* IIWI INF* (I1F INI IN.t N ,11 m. MN.V9IB 7m -TITIlCHLMMINNIirUMm mm TITl(IClHiNU|^<|1|XiM IN 1 NT IMNII FtN UNNIMIU DPil* SINN IN IIMNMTM Nf* Ml tlIN PNCtf ( 1 1 i?lt*Ffi, IMI MLCN 3 .IT NL U-M HfltNHl IM MUINlMTlMI M CNLtMtlttNtt-FIM1M IIMCH IN hlVKMUTM FINN CMMtMIH MMM NtftfNTKt IT ttfN MMBN NffC 1 INN, MKftltMl MCI ITT IN MM MtCT--NCIUMINMMIIC, IMl M K 9 XK M IIKJM t4 t MMKLIMt.MM < C WfNMI MiimiM m Ml IMNMTM LHI MHL It tCTlMTt TM Ptttlt- TfNCJ tt CMfMlIMflITt IN LtNtlC fN- TIMMUNTt IN MUNIKC TMKMLMV mm mimm chunikm MtMm mm 7t mtn in im,mmi- CMI MCICTT rm IttVIM MM MftMNLt iNruNtiNlltllll PP 114-lir. K 1 tNLlINC.t t-.tt M. IMLftTItN MM NMLTtlt m HLIOlMlMlffl tlNNit* MMN in Mine tMMlCI. 1Nt- CNLt* tlNNTCt tltNtttl MM MilTCt CM- NHMCt INHCI Ml fNff UNIINNINT, t WlllMlltll M. tKH MfMMI Nfll.CUNIM.C v , IM| M 17-M 1> II IMl INC ,t l .CT M MtlMNK CF PMVCNLMIIMTCt tIMNSt-F-tIMIKNi MM tlttftWlMMM IN LMMCNTIM MINI IMItC Mt MCfCNtft CT M INICtMTIMMi NHMM Ml CM.MIIM- rit CIMIM MM MLNfIt eMMMMtt, MtlNtrM.NIMlNIN.tCTMCt Ittl n it a i F-S7 HONS 21S375 Nfcj*iN or icrtifNCti o*o ret* .-KftllWLt ITAML | J I ,r N IWilNC MO MO* LVftll NOOtOCOL !> AtOCtOlNO OMMlC CNItllMt * CM CIOnOMAiy COMOVtTIOM 140NCC1 IN tNNOWNTC SOMIMTlM 0IV1IIM COMOlHtlOM tTUOItf K00*900/9* 0J-4M N100C4T OtlCOftCH IMVtlTUft, MUI Cll* NO JNMQOT INJ to- felt Mint XN f I |NL N MO L L LMFMtM 1 Cldll* MOM or OCVCOOt l.t.HmiMMIK- MONT tOWOlNM* 0MM09IW 00 M,M- rftfc*CM.M0910tM90-9-01MfN OCItMCC ioriitoe*!loo,itir x IfML t I .(I N. |Nf ilMILITV m ffUTM* IMWClM MHO CMLMinortO 010MIMC 10 OUNI.IMT, Nf*t, MHO CM* 0U 11OM iHt CMLOMOIMIMt * M1CIM mo mu c n olnio,ko Mu cm oco ,ictiiio*mriori * IIOIK.O JIM* TCM MO COOMIC NCootic ooooinroio iii cmlooiroico oi- ON I MO MO OCLAICO CMMMOI IROOCI on me twvmmciiT o wttiMfMf ML ,KOI OCOflMM Mtll^CLOMMM V tMI M 010-9*9 OUMOtTOM 0 1.9 4.4 -TCTOOCMLMO* OJOMMICNL two 1,1 `,4,4 '-If TMCMLOM OlOttVOCMH M FOUNT CMLMACtt>OCH MO IMlVM IMOMCfOO M OVftOIfU IN. CMLOOIMNTKl1 010NIN0 ONO OflOTCO CMMUOI JUTACT on TMC mtOMMlNI 0 RUIWII Cl Oi .coo OCOCMOM OOCtt.KLtttrooo.N T >000 00 999-199 in cmcM %/t lew rwt Inv urc mm. wn muh MIN AC CP INV TM Ift 1' MCf CMM CMCH CMC* CNtN CMfN iHCN CNfN CMCN TON TON INtft |NO I INFO >11# x M IT N N N M N VH BS-J HONS 2X5376 Nr*r* of unacKCi pm Pdf HO- (ui- ctn cmtm i/r tera awr inv nrc mum. hyii HUM (MIN NfCO (MV tPM fTP(lf MINI *M| NC CNM CHIN CNffM CMCM CMfN iMCN CHtN CNCK to ii imct ioi INF0 6 ISP rVLM J I ,MN I M tilvf CMLMMCNC PltM 9,1 4,4 *TKrlCIHMM|VV||M- 1KMC mm M'.tr4'*tfriMM.aMMMH- 2(Mi WMII Ml MVItM 1M| MMH- MTCC IIM1M mm llllld CMHII, IHFMT M INC CMlMMtNI Millie CSC,SI N .CO PIHMM HCM.tiM- PMI n V JMI M UM41 UUM.N t tIT M. MNHIINri M IN* CIMiNTICM HMIMCI M MMCfl M IMCttlKIC CMLMINfctCC NVMKMMM( CMCtNlHTICM m CMI/CMI NN PM' n UMI tCCMCCC CMNMt IN INCH*** ^ to cfplvcnti PMimrci it tic c> CIOIN inPMlIM.MIMI CtlV.IMaMN* I.MMCNMi IMS riVLMJ t .(1 CL ICVCIC N 18INN* CNLMC-CIMNJC-C-+18N1N IN IMINtt- MSNTM. mm SlClCCICM. OMNI11 II N* tCMIMI NT CM CflMMfttOCMNNt-NItN UMttftlNI MM MCTINCtlV PMMNtCS It S4TN MMM NCf VINC.MICaiCNN CM 18TV PM Mil iPCCTMUtM.IM HICK, l*PC TNJMMMM.L 4 MP-I1TC IlMCPMt f t.J.P.i TC1iMNLCCM1tMfC~N* CIOIN FCCN ft CCCCNCrICN-'tlIPMN rcciuiv MCMNrcc ct tnc mi cioin VRPM1MI IMCM CirtJlIMMI.MP* tract ini IICMHMI, t C CML8MC1MNSM1CMIIIC mm CNL--CIMMCCCNCCMCi IN MfMllN iNi ccimmcciiM n immimi KM ICC * (NNfl.ri mm NCM CC18NC8 M MfM N1CN1CftM,1Mt PP MMIt , M I l 1I I i l K * MIX k HONS 215377 Mi*TM ar acFcacvcca too rtii aetmenh a/raia* tva* aiN cncn tint ahoi cnw nfc anal ncia nun anin acca cnv mut tet/ rca* MINI ANt HC CHAO CNCN CNCN CNCN CN|N CNfN CN(N CNCN TON Tan INC* INTT INFO MSA 1 IIIMW< T t ,CT AL MMlfTIUi NCW- AM X MKHT FH HTCMJMTKW AT CIUH- iiNNmtiiriM ana cmwinni* ruaava in nv am mm irtei iFriMin FAAN NWNIC1AAL VAC 1C HtCtHCMrtM ratacNTta Nrwc tnc aivtaia* ar cn- VJAOMNCNTAL CNflliTIV.MMlIM CN>lK-> cat tacitrv,i*a vtftaa.aaacN im rifaMAM.T .,|1 AL CMNCrCarMTlM ar tank caarMCNTC in rut imimti raw a acrvM-riata latiHraocaa, alum caaev..ei *o-*A4,iaa. Na a 1JCANAN,K | ,Cf AL AIMIM. ML.t ANALYTICAL NfTMA FM iMMlftlAL MiTia t*A-4t/t~fttit?, MtatAL KMiaanwftal NMMO lomaimt. C1*tHM*ri,MM IMI. * * TlfMMN.T a JT AL. IWIMIITIM AT ANN K * CNCNICAL iMTH CMTAWM NIVCNU* aiiMitt inianit Maiavnr ar naff eauancTga at Hutu cmvimnmcnial atavieca, acn race, rmi mm cncacy iiiim cm*any, l a*a*a, AtKAMaa ! attatncAfiw ar MtAAMM AAtVSC 0 KNNCa.Ca. ANN aoaaa k iimc, mm aaaM,viciiiiAM, IfM 10-1*4 laaiNf.M aiHiMi a conation r*irccrrvc tmkntc* at Tnc aca aiania tvnaafliaa eanoao crtv.aiiiaiiMtF* fsnaca iaac. N lawnA a i me vac ar aiaaia car* an vn# aaiim fa tacntity aawcca mm active aacactavva Lcvcta ar aim* ina in tnc cavravNNCNTiA Btvitv, oavc, mm cmaaiM ar tnc racacvi riaicf in. CHLaainafta aiMtva ana actAfca canr#wiM, intact an tnc in> TiaavNCNT a nviiinvcn^ci al ,caa rcaaAMN racaa.cmaraaa, n.v ,j**i rr.taa*CT4. N AN MOWS 215370 iMitm ar iirinwH raa ^ci vriKHrr ail* rut* tin own i/i rent Mir car art mmc hci* hw miin tee* inv i*n t*t/ ret* lint Ml NCf OMt OWN OWN OWN OWN OWN OWN OWN tM Ttl iHCt INNI IHf CM f TtMM,N * ant a ttmm. CMtarcai nnn* CIIMI IN IHf IKMMIV1W NT MIMt* M MICIIMil MNltH KINffTIC* #1vmiM.ctiifCt rat ownjcm turtle** MtTItNM. tWNfNH tt ITwMM.MHlM-> T#N,t C N N * VMtHlNNft.t *r MTWWM.Nt WH Mil m MtM^TCN IN Ml WTMW MNt MW IN NML ttatric ItMTITIW tNVlNMt ftHKttt OWN ali4lM.1M. * na * IMF N 1., MW t MFTllMtt* MT Hf IN* MLltN tt NttTCNLMlNNTtt IKMNIM* tltNINt CWMWMM, Mil *IW. If?*. K NR rWM f M KNMIU'MW t.NNfriH! INC MCftitaa. Nffaa&fat W 4.4' aicHLtNaaiMWNVi mw itk Ntmutw NT MTtMMTIM CNNflN tMNCtC, OWN*' ttfWM. 1110-1 at, ItN. a n rwwai n *aab i j cbojim, iiiiwN an van aravt arrm aaitaiM awro NaamaLaaic NiTtaariaaw in tiw iivcta ar aaiNta rtta him rate a inti Nii er HCM.rN.MNMnr, itaa am u a CHriNaMWNrat mrccTian aacncr cairctia mw hnwini tmtra*. a. a,, a*igTiNcwtaaaa iaaui*t*ai m in MWICUT WVCI tmlTI CtTTCffla. NNaMiNtraN.t c,,i*a w nu u a aur laaMWNfM. raa reel ran ncchcv ctiraau mw arwwaaai ainaiaa nn* aiCNV NNTca MM.MT cantata raa NNircHLaciiwraN tirwwia (^a/i- aa-ata.NNMiNtraN^a c .acvaaaa itaa 1 MR F-61 HONS 215379 Mfiu 0$ hfcKmii rai MriMMCt________________________ VM mitft.J 9 ,4 4 UilCM,M J,| lcin rnrasaafa mciMitcc at m+tinam in ante (iHin ?a ian Lima m 3 >, 7 J rKTHCMM1Nlllt*^ blOBIN It- fvfl- in CNCN 9/1 TCMt PMr INV RFC MMK. Nfl| MM Ml KM MCC* tat Tana ri/ tca MTHi ni Ml CRM CNffH CMl CHt* CMR CHCR CNCM CMH w rH met (Mff 1HF0 MIR Sa VM R1UCU 9,4 4 LUICR.RRI FR MLLCM IMCtfMMa 1MCIMMCC W Ml^HNI IM Mil IRMWI ft LM iCVSLC I I cue**a*MCM, a*t'ija,ia? r VMM(llrS r ,f ML ICIIMHIIMIIIIR' 'n za-R-aiMUMi IM CtMRlCML RMVtl, o* nautana ctnuvara mm mu mmm- n IM|IC,Mai-MMNl VICCNM ,ff ML CM7MIMH Mt 7MC iMWMi#Mactaivc crasev at a,i,?>a~ TS CanCNLMM tNM2#*M*t ttHtV t fCMM V MM *,l,7,a-lIMaCMLtataiinf-nMMI <TCW> IM MICt MMCBtMtCa If 7MC MM attain ivwiiUM.iMRai city, miaaavi,aitnnaaa itta K I a VfCRCtMiul.tAt r.1a1zM.aL rT(.FtfflCTTMaMtCMMCLMfafla|iR>t*tyf> t-aiamn an mmmtu. anuaaav ttatmnan in met cntn.afaL inrtantftana, iiijiMii, itaa. R vftmMMM,a mvvch. mm a narnnaca me nciMtat ian at aant cntaainmca aiternatiM** at aara, cncnaatMta* 4li3l7->a3 1MI. a vcntMt.inc Miraaccananic inane ra at rnc taaaaca tea an* attaLMnana iM Ma/i'i7>a)MniRBiiKia,M ,mt ttra a R as MONS 2153S0 PM' ir. a OF XI iii> m- ITP CMP l/r IKHP Hit I IRV RPC MMK. MTB MOT *MIN CCB (MV CRM Til/ KBt RIM Ml Cl CUM CM* CM* CMC* CM* CM* CM* CM* in TOT IRC* NPI IMF* HIP I i i,r>rirmtUMuiin< PMI'IIMIH) iritcil *M MCJUNftM ini cmlMinricb pm--rv *citi mm IMia B1BR1M C MKL.n cc*t ULl I IMCRMt* >, I? IIMU.mi. * ROB 4 S mm 4 N INMW CBMMflfM TOItflLMIC BIMV *|T* MtMLlllMTtl IIMimi I* CNIMNI *11* MCI* ICrtHWl I* MMWItl, CM** NRRA- n*. Live* mcmi* mm TiacM wbi- 1*M* IttlCCL *m . POT***C*L will |U<4i*1?l I I *01, J C ,(| *L INITIf 1C*ft*N mm iMit*i*cic*L iwinrin m cmlotin*is* *tKWMMi mm c*l**i**tc* MPNTIMLCM 1* Nl COTMM1H P*LT- CHLMIMIC* *l*MltVL*, PM# C**Mf. f**1C*L ,*i-IMm I RN HI,! a J 4 Mill,** 4 C.JKML IPfici m B>.T,*-fT**fi* wMinmtPa*1tttR m TM IBM CtCTt* m L* ******** M1MLB ClfVI*** M*Lf* MB* BPre , tKf fRIMfRI *L IBBM SiHt-tll. i*n I m. JB ,4G rMCriCMCM.BM* L fftTB* 1NNOTC MBMIIIIN *v TC** J*t *|*M|Hi l**l*LMiel M CM* ICut MNCIB r C*1VMC*I.* C*V*U*M.*N S Mil Bt* PCIBP MMLlCMflfM*. MU VOTIr1BF* PP MR* C f ,*M t M*TBVHUt* Mt| BF |r ]r r. B-TKT**CMLCMftt*fHJB-F-*lB*IM <TC*B> IH NBML AMlVIC BNVVM*MNI A*fR C*Vl*OT CBM** TOTKCBL ,P> R * R R R r H MOMS 215381 MiftiK or tcrroittcso roo rco# liftrath------------------------------------ VClfWM L .MO L NICI, OJOTMINt MMkv or rcoai WMf OVOILOOLC. MOT OM TNf Ml NHI,1<|)| ra'M,toot. oio- rw im coco o/t iff* mt im ore mm otro MM AMIN OCCO fW (IM 101/ wo* aiot mi mc com chin coco cocn euro two cot*-cota tm t#m mca imft iiri oil* nk MatIM O N 0COL00 teal CONOIOfOM- aanivttiomo ikmiiio cnvimonnkittm. contmojmotiom aim M.rxotia- CM.0O0010CMrO-M#*-0l0fN IN* CtO.0- a j motto ropmoot ocio# omo MIO 010- tMIM C OOMCL.CO. aCOL-MU lOfOCC* tm.UK ir/tN-m.TiM. OKIIfaiMrT fNff MHNUi OMO no ionic clooo vou MiffMm mca, offv mooch.como . iora. I kak OMlTMOMt.r C. 0 MOOT |MH MOM OOLAMCt HOOCL W TM> OMMtfO, 010TOIOMTIOM. OMO TOft OT KM 10 OK OHoioiMMiir, m-MOrt-rr-ooc.ocMoa INC ,OMO KNOT J1L0,00,JOLT |0FF. " OK MlITHIM, r C MOTMPCflOM OT MOLT* CMOOINOTCO OlOOClVTLO IN OCOMOt OLWOttf OOOIM INC MOOT TOM mm. inc .oooaMorifio.oo..tore I aiLUOMt.o l .ooo Q.PLMOritiO. UMCNIM MTMOO 000 tOt OCTICTSON OF CNLOMOlOCOfO-O-ftlMM IN TOO FOtSfNCt or CMLOMOOlFMOOria. COLOOOMOFMIMOLCOCO. OMO COL --NIOOllTOrOMOMO. i. oooc orr omul cmcn ,os*oj-oo,iof aaa OILOOHrO c .OMO i T ON | TO 00MOIM. NCOOOOta rOLLOOM MCCIOOOTOL BSLtAOC or OIOMIM TO 00 MCOONTtO NT fNff JO INTCNMOlIOMAL OTMTOOION ON MOOlNO- ICO OIOOIMO UNO MfLOlfO COflTOMO, OOLIOMOO^OMOTOIO.OCTOOCO 1000 a V a a HONS 215382 S9*J Mini or NflMKCl r Kh io* rut' TO CMC* 0/T TCMT MOT SV NFC MML W10 MMt MIR OCC0 CUV liti in/ Klf KlNO MM NCt CUM CNN MM CtMfl CMCn CHIN CNtN CMCU vm rot met ttr I oiir <Mpr uc * ti oc riu.0 toim.0 on MtlMMMNI IM or KM ON Utfff*TION OTIC* IMtTlM NlTN VfttTOOLC tit in nuiin iMicnintM mm CNCR1M. reel*. f.CftfTOOCNl. I. COVM.COM ,M* O.MILliM#* QMtTtMl MtCICOtlOMO.INN HU, 1010 rr cot-jif, WNt,0.S MVCMIMTIM or KM ! iMtmiOt MM MMCfNC UMUMim. IPMMN*N-M.CIilMMlO OMCTtlCOt itMMIMV,MCMflMTI.MIL JM|. hmmmchcut non rot oiort or taLiiami* roc veil ico nhi roofcmco ot TNf mi ret OCNINOO.OOlCM.TtNOC. BCCfNMO I ftI UMLOOU.C O.r0 r INMi.MI III N- too mmvcv or MiftMOMfimut^r* OIORIM CONTENT IN OtCCCTCO FOOlI- cioct o oftic ia mnmH<nmn. ion K VMMMIJ .V VOffl'MM MINN* UTMOINKMOL ttCNMfOIIINl OT MtCMCMtMOlCO OIVNMIO, CMfltft Kt. ICCMML tlOiCtt-ttt, 1000 * I * I HONS 21S303 mill i ar UfiitMCi raa ki it- fim- vn caca /! it at fmot (f *fc mm. ncti niir mih *cc i*v ran* nr/ Kt HIMI Ml Ml CUM CMCM CMCH CMC a CMCH CNCM CMC* CMC* ll Vd IMI IMfl lFt |I|F TMCJ C ,Md A S RMLMM, M1IM Ma- licm V CNLttlNlblfC IM IRi CNLflMlMIM - MICIN M MTf C M K*ia,K caca mi , laitiH) ifM, rcctiM a , a. lacMic ,* a a jQnca aiaraiawvvan aid ramci tt it tcm ana m,m in an aaaaiic hml staavaica cwfaaaMdtt,yaiacca. im VdlllMM ,cv at INMCflVC tFTCCr an ncmtkc uuvata aid imik Taaicm ar maiviauai pm.rtm.rnimiu tiata* s MFM cnmnvh ta mi, ramcat. am FMataacit . a*i*aa-iaa, iaai raawiMM*. t. ,mm a him eaaam ar ichni iNutata vita mvcataaiMrca UMim raiaaamt aa naa cariaa** tea it ,c i imtici.Mi a j caiMv Maaiciat atonal airs ratalit mm fMiaaiaiciiTat aa*itaai*c. ra-r*-iaa,v a aia raaet accaaailann mi taviaaMMarat idat ra taaaaataav. ataara *rt,Tc**a, itrt imnj i idiMMntai cafamtv mm caviaaancarat laaicataarp acvicv tscTutc fa at NfNiiti at rat aaa mrcaiMiiMMi tvmati an aa cataaiM* ira aiaaiM mm artavta camndt. annate mania,acraaca ita* HONS 215384 nitaii ir acrvMNCfi pm .JKUIIHCL VCMMC * L INC CN.H1MVKI 01MN1N*P* MOIINI IIP TMC K1CMC M CNC CCMCINTIC PNCMCKT IlfMIClNC N CCVCV MN t TMR.CM 0111V. NC0 T0M.IC0I PP OMN YMNM,C l ,tl Cl INC T0N(tCLCtT,fMVIMNNKNlCL MIC Mft H0NM t|M CP NC001C10C HMH M0 |T# NCCCCtNTfl CIM10 TI-ll-tM C 1C PNNCC CCCCPCI10NM NNC CMIC0NNP0TCL NCCtPN IC0MCVMT,M0CC NP*,TCCCC,0C?0MN icpc It- PCN- IN CNCN 0/1 ICNP Ml CMP ME tMl MT0 MUR MIN MCI (MV 1IMI 1CI/ Kll KIM Mt Ml CUM CNCN CNCM CNffM CMN CNCN CMC* CNCN W 111 (Ml INPT INFO CUP Rill MM V0UN6.C L ,0 4 CCI0N(1,M 0.CNN0- MU PCiCICTfMCS. MCCNCNT Mft M~ CMTNNIMT1N CTMItl CP TCCC 10 CfCt* ACC CVICt MCCIMLT C00T00IMTU 0IT0 PMNOKV NCMVCtBCC It N PMCCNtfC CT TMC ICC INTCNMTlCMNl lYNPCClMN CM CMLCCKMCICC CICNK00 M0 HlITU C0H MI0lC.CCI0iC ffCt. IMC.I N ,M0 N CMC INC TMC MIC* HIT CMPCN1CNCC CP NM(N CNPCCCC 10 ICIiCMLtPC010tNfC0lCNlN IN N C01- CM.C00PMC0CL PMCC00 KCCICCNI MM, J CCCNP Nf0.,00*11-14,I0C0. CTIK0 * PCL VCNtMINNICC CIPMCMtll M0 MIIM0CMLMINC PC0IUI0C0 IN 0000 nCCMMTCN M0 NMKNC PICMN. C0U CCVI0M CCMT4N1N cpi'tert ItllC.f ,0 N0TIIMC0.M0 P N CMCI CNHCNIMIICN 0P TNC CM CP P0M0YC0LP CP MINC CMC CltC PCLTCM.CNI NCVC0 CIPMCNTlC, PCLVCMCNlNCTCC ICt- PNCMTLP, CM. MlNATO 01BCN200ICN1MV CMC C1MNMPHCCM. CWPICM.NCCLTC PCCCPCC rlNPCilNCttlM I0C0C 1 r4P*0i, im RK HONS 215305 89*J legend PCBt DIOXINS FURANS OTH HCS CHEN CHAR O/T CHEN TENP CHEN PHOT CHEN ENV CHEN NF6 CHEN ANAL CHEN NETB CHEN HUN TDX AN IN TOX ACCO 1NCO ENV INPT TRN5 INFO TRT/OISP Polychlorinated hi phony Is Polychlorinated d1henio--dio*i ns Polychlorinated dlbentofurans Other hydrocarbons General cheolcal characteristics Deconposltlon/transforaatton cheoistry HIgh-teaperature chealstry Photochealst ry Envlronaental chealstry Nanufacturlng chealstry Analytical chealstry NetaboHc chealstry Huaan toxicity Aniaal toxicity Accidents and Incidents Envlronaental Inpacts Transforaer and capacitor inforaation Treataent and disposal MOMS 215366 EPRICS-3308 Balow at* flvamoa* cards that crowH*tffc*t m addition to allow tha utta tiling ot tfts according j rtoort. a txut abstract jMctlbinfl tha major subject aim conarad m ms raoort > mciudad on ! :I tr s!* f: 9* i: vtCTbic 0#M MltaACa uitHyti n ca t*Ma hi hm CNUCS4M Statfroftho-Art Ravlaw: PCDDt and PCDFt m Utility PCS Fluid Wiw c* I antf Bmr nwumn m* iwt n.t. U r.** rs taimticOihfmticata<*4't iamnaniaituieMRW imiriMrluW*Wc i* EPRIs* Rptm-ti i: Stata-of-tha-Art Aavtaw. PCDDa and PCDFs in Utility PCS Fluid v. I: MOHS 21538?