Document gOvaaYBXG6XBgyg3rK32V0qq

TO: Distribution Int rofflee Communication FROM: DATE: SUBJ: T. G, Grumbles August 28, 1991 EPA REQUEST FOR INFORMATION ON EDO Attached is a memo from EPA informing us that they are concerned about EDC emissions and exposures and requesting "any additional hazard or exposure data .... that would be useful to the agency in more fully assessing the risks of EDC." This letter is from the Office of Toxic Substances (OTS) and is a result of a new effort by the Agency to review TRI data, identify high volume chemicals, screen for multi-media exposure potential, and then more complete risk assessments by obtaining data from industry and/or requiring testing be done with their TSCA authority, I'm providing this for your review and am not proposing any action at this time. I am currently working to determine who else received the letter, determine if trade association (VI or CMA) action will occur, and to get more information on this new OTS initiative. After the above is completed, I'll contact you regarding our next steps in response to the letter. Please call me with any comments or questions. T. G. Grumbles dlj .313 Attachment Distribution: David Booth, P. Carrico-LCVCM, M. G. Hayes-LCCC, D. Penney-Austin (EPA Cover Letter Only) T. H. Huffman, R. D. Gamblin, J. Friend UNITED STATES ENVIRONMENTAL PROTECTION AGENCY WASHINGTON, D.C. 20460 OFFICE OF PESTICIDES AND TOXIC SUBSTANCES Mr. Thomas Grumbles Manager of Environmental Affairs Vista Chemical Company, Inc. 900 Threadneedle Houston, TX 77079 Dear Mr. Grumbles: The EPA Office of Toxic Substances (OTS) recently conducted a preliminary review of readily available data on the chemical 1.2- dlchloroethane. The purpose of this letter is to inform you of the results of OTS' review. Background materials OTS has prepared on 1,2-dichloroethane are enclosed. The Agency is concerned that there is a potential risk of cancer to persons exposed to 1,2-dichloroethane. 1.2- Dlchloroethane has been classified by EPA as a "probable human carcinogen." During the OTS preliminary review, possible routes of human exposure were evaluated, including workplace exposures, end-use exposures, and exposures resulting from releases to the environment during manufacture, processing, use, and disposal. This multi-media review indicated several possible exposure scenarios which could contribute to a risk. This review also indicates that more careful consideration of 1,2-dichlorethane by EPA is necessary. OTS will therefore continue to assess exposures to 1,2-dichlorethane and consider whether and what risk reduction actions may be appropriate. Therefore, the Agency is taking this chemical substance to the next level of risk management review, where we will address hazard and exposure Issues as well as pollution prevention. While OTS is pursuing this more intensive assessment of 1,2dichlorethane, I want.to encourage your company to undertake voluntary pollution prevention measures within those parts of your operation where the manufacture, processing, use, or disposal of 1.2- dichlorethane is performed. Pollution prevention is the use of processes, practices, or products that reduce or eliminate the SAL 000066780 Printed on Recycled Paper generation of pollutants and wastes.1 Because OTS has not evaluated specific details of site operations within your company, EPA is not recommending specific pollution prevention measures at this time. As a general matter, facilities are in the best position to identify pollution prevention opportunities that should be explored. As your company may well be aware, this is not the first nor the only pollution prevention activity being undertaken by the Agency at this time. EPA, through the Office of Toxic Substances, is also actively promoting a voluntary effort targeted at reducing the emissions of 17 TRI chemicals through pollution prevention opportunities. Your CEO has probably already been contacted by the Agency with information about this program - the 33/50 initiative. I have also enclosed a copy of the recently enacted Pollution Prevention Act of 1990, which outlines the pollution prevention goals the Agency will be pursuing in the near future. I would especially like to draw your attention to section 7 of the Act, which details the data necessary to evaluate pollution prevention activities and plan for their implementation. The first reports under the Pollution Prevention Act are not required until 1992 for calendar year 1991. However, I believe that immediate voluntary implementation of all feasible pollution prevention actions would be prudent. I would like to offer you the opportunity to submit additional hazard or exposure data on 1,2-dichlorethane that you believe would be useful to the Agency in more fully assessing the risks of 1,2- dichloroethane. I would also be interested in learning of any actions your firm has or plans to engage in to reduce exposures to 1,2-dichloroethane, or to implement pollution prevention practices. OTS will incorporate information submitted by your company in further Agency"assessments. " 1 Within the industrial sector, pollution prevention is often referred to as source reduction and, according to the Pollution Prevention Act of 1990, includes those activities which "reduce the amount of any hazardous substance, pollutant, or contaminant entering any waste stream or otherwise released in the environment (including fugitive emissions) prior to recycling, treatment or disposal. The term includes equipment or technology modifications, process or procedure modifications, reformulation or redesign of products, substitution of raw materials, and improvements in housekeeping, maintenance, training or inventory control." The term does not include any practice which "alters the physical, chemical, or biological characteristics or the volume of a hazardous substance, pollutant, or contaminant through a process or activity which itself is not integral to and necessary for the production of a product or the providing of a service." SAL 000066781 A copy of this letter, as well as the background material on 1,2-dichloroethane enclosed with this letter, has been placed in a publicly accessible administrative record for this chemical. Please send any responses to this letter to: OTS Document Processing Center (TS-790) (Attn: RM1 Process/1,2-Dichloroethane File) Office of Toxic Substances U.S. Environmental Protection Agency 401 "M" Street, S.W. Washington D.C. 20460 All responses and correspondence submitted by your firm will be placed in this administrative record unless confidentiality is claimed at the time of submission of the information in accordance with the procedures outlined in 40 CFR Part 2 Subpart B, and 41 FR 36902, September 1, 1976, as amended at 43 FR 40000, September 8, 1978 and 50 FR 51661, December 18, 1985. If no claim of confidentiality accompanies the information, the information may be made available to the public without further notice. If a claim for confidentiality is made on all or part of the provided information, you must also substantiate the claim by submitting a full written response to each question posed in the enclosed item entitled "Support Information for Confidentiality Claims." EPA will determine whether the information in your submission is entitled to confidential treatment and such information will only be disclosed by EPA to the extent, and by means of the procedures, set forth in the cited regulations. Please note that failure to Include the responses to the substantiation questions for each claim of confidentiality will be construed by EPA as a waiver of your claim. Should you have any questions on the information contained in this letter, please contact the TSCA Assistance Information Service (Hotline) at (202) "554-1404' for 'further information. The Hotline also has a package of pollution prevention guidance information available upon request. Sincerely Mark A. Greenwood Director Office of Toxic Substances cc: 1,2-Dichloroethane Administrative Record EPA Regional Offices Enclosures: 1,2-Dichloroethane Background Materials Pollution Prevention Act of 1990 Support Information for Confidentiality Claims *f c r m a t; ; - . "2 - sceciflc rsccrt ir.2 r? ; . 2_2 - -. :: - 2 T:x ic Fucsca-.res :r:l 7 = C A i :r in ::' 7 3'.- :5 ric'^ct to the :r;visiDr,5 of 3ec cion 14 of 7SCA 2". d is E?A'^ "emulations on one Confidentiality of 9usiness Inform a t it n . ;ee 40 CF3 Part 2). You must compLy with the following procedures to assert a claim of confidentiality for the information solicited in the attached letter. failure to follow these procedures fully at the time you submit the information to EPA will =e interpreted dy the Agency as a waiver of your claim of confidentiality. Asserting a Claim Information claimed as confidential must be clearly marked by boxing, circling or underlining. All pages containing suer, information should also be stamped "CONFIDENTIAL". Care should be taken to ensure that these markings do not obscure the suo- mission`s text. Sanitized Copy Two copies must be submitted of any documents containing information claimed as confidential. One copy should be com*plete, with the information being claimed as confidential marked in the manner described in the preceding paragraph. Th other copy should have all of the information claimed as confidential excised. This version will be placed in EPA's Public Files. Substantiating Claims of Confidentiality Detailed written responses to the following questions must be provided at the time you submit information for any portion of -- the in formatron-you--claim-as-confident-Lal..__ YQjjr__r.esponses _shoyl.d_ be as specific as possible, with examples as appropriate, and should provide substantiation arguments for all types of informa tion (e.g., sales or product ion/importation volumes, chemical identity, company `identity) you claim as confidential. 1. For what period of time do you assert this claim of confidentiality? If a claim is to extend until a certain event or point in time, please indicate that event or time period. Explain why the information should remain confidential until such event or time. 2. Have there been any confidentiality determinations nade by EPA, other Federal agencies* or courts in connection with this information? If so, pleaae enclose copies. (over) 0000i.83 s^\- 4. Briefly describe any physical cr procedural restriccicns w i zr. m ycur company relating ca the use and s to rage of the information you are claiming as confidential. What other steps, if any, have you taken to prevent undesired disclosure of the information during its use or when an employee leaves your company? 5. Does the information claimed as confidential appear or is it referred to in any of items listed below: - advertising or promotional materials for the chemical or the end product containing it; - safety data sheets or other similar materials for the chemical or the end product containing it; i - professional or trade publications; or* ' - any other media available to the public or to your competitors. If you answered yes to any of the above questions, you must indicate where the information appears and explain why it should nonetheless be treated as confidential. 6. would disclosure of this information be likely to result in substantial harm to your competitive position? If so, you must specifically describe the alleged harmful effects and indicate why they should be considered to be substantial. Also, you must describe how disclosure of the_"infbrmatlbri would 'cause " the harm. ~ 7. if the information in question is "health and safety data" pursuant to 40 CFR Part 2.306(3)(i), do you assert that disclosure of the information you are claiming as confidentisl would reveal: a) confidential process information; b) confidential proportions of s mixture; or c) information unrelated to the effects of the substance on human health or the envir nment? If your answer to any of the above questions is yes, you must explain how such information would be rev aled. q0 RM1 Meeting 8wn""^~v Data: June 21, 1990 Subject: 1,2-Dichloroethane chairman: Jim Willis coordinator: Nancy Chiu Supporting Docs.: RM1 Briefing Package on 1,2-dichloroethane 1,2-dichloroethane (DCE) is recommended for follow-up in the STARR process. The major hazard concern identified is carcinogenicity. It was recomended that DCE be added to the Risk Reduction Candidate List for Occupational exposure, and ambient air exposure. The worker exposure value (81,227 workers at 2,154 plant sites) was questioned. It appears that this value includes all of the work rs at the facilities, however, it may be that only a few are actually exposed. The decision was made to send letters of concern to those facilities which show the most significant releases, using the same criteria as in past letter cases. * The decision was made to evaluate optional criteria for determining which facilities will receive letters in this and similar cases. This analysis should be performed by the Existing Chemicals Assessment Division (ECAD) and the Exposure Evaluation Division (EED) within six weeks, and a paper summarizing the results circulated. The case will be brought back to RM1 in 8 weeks. It appears that there is there is not enough data on Immuno ,,tox_and_. hemato- tox. `"HERD should examine these areas. The effect on the Margin of Exposure (MOE). for the most sensitive endpoints (othe than cancer) if controls for cancer risk are set in place was questioned; such controls will raise the MOE to >100. A decision was made for HERD to look at the mutagenicity, and to determine whether there is a need to go to Tier III. The results should be reported back to RMl. SAL 0066785 I. SELECTION RATIONALE 1,2-Dichloroethane (C2H4CI2, CASRN 107-06-2) was identified during TRI screening as a chemical with very large production volumes and environmental releases. DCE is toxic to humans and animals if inhaled, ingested, or dermally absorbed. It is classified by EPA as a Bl "probable human carcinogen" based on sufficient evidence from animal studies. II. CONCLUSION AND RECOMMENDATION 1,2-Dichloroethane (DCE) is recommended for follow-up the STARR Process. The major hazard concern identified is carcinogenicity. DCE should be added to the Risk Reduction Candidate List. in Based on the 1987 TRI release data and the chemical/fate properties, the exposure potential of DCE to environment (air, water and soil) and general public is estimated to be high. Preliminary quantitative risk assessment, using the environmental model estimates and EPA established cancer slope factors (inhalation and oral), suggests that approximately 66 and 83 excess life time cancer cases can result among populations being exposed to DCE contaminated air (72 million persons) and drinking water (40 thousand persons), respectively. The highest daily inhalation and drinking water intake estimates are 0.06 and 0.022 mg/kg/d, respectively. Limited animal studies indicate that possible immunological and hematological effects can be resulted at these exposure levels. Additional modeling studies can include verification of site-specific model key parameters (e.g. air emission stack ^ height, actual release days), intermittent/batch/accidental release scenarios, removal rate during drinking water treatment, ground water exposure through leaching, and acute aquatic toxicity due to DCE releases at low flow conditions. Testing recommendation for immunotoxicity and blood effects can be made since there are very limited data in these areas available in the literature. III. BASIS FOR THE CONCLUSION Chemical/Physical Properties DCE has molecular weight of 98.96 daltons (conversion factor 4.04)and is in the form of a colorless liquid. It is very soluble in water (8.69 X 103 mg/1) and organic solvents (log Kow 1.48) and adsorbs moderately to organic sediments (log Koc 1.14). The Henry constant and vapor pressure at 25 C are 4.5 x 10'2 atmm3/mol and 61 mm Hg, respectively. o00066786 2 Exposure Aspects Summary Production/Use DCE is produced commercially either by the direct catalytic vapor or liquid-phase chlorination of ethylene or by catalyzed reaction of ethylene, HC1, and O2. Production in the United States has increased from 7.3 billion lb in 1985 to 12.9 billion lb 1986. Importation of DCE in 1985 is 14.3 million lb. DCE is mainly used as a chemical intermediate. It is also added to leaded gasoline as a lead scavenger. Approximately 84% is used to produce vinyl chloride. DCE is also used in synthesis of 1,1,1-trichloroethane, aziridine, trichloroethylene, tetrachloroethylene, and ethylene diamines. Other miscellaneous applications include use in solvents for various extraction and cleaning purposes in organic synthesis, as a dispersant in rubber and plastics, as wetting and penetrating agents, in ore floata tion and as a grain fumigant. Occupational Exposure The 1990 NOES survey by NIOSH shows that 81,227 workers at 2,154 plant sites could be exposed to DCE. The occupational exposure areas include food processing, textile mill and textile products processing, chemical/petroleum/rubber products production, primary metal industry, electrical/mechanical and transportation equipment industry, business and health services, and museum/botanical/zoological gardens. Between 1979 to 1990, OSHA documented 41 screen values and 105 OSHA personal values. Among the personal values, one ceiling value of 160 ppm (at J.T. BAker Chemicals, N.J.) and one TWA value of 59 ppm (at Plastics Products, Texas) exceeded the OSHA recommended standards (ceiling 100 ppm and 8-hr TWA 50 ppm). Environmental Fate DCE is very volatile from all media. The estimated halflife of DCE in air ranges (via photooxidation) from 12.2 to 122 days. DCE is biodegradable by aerobic microorganisms in water and soil with estimated half-life of 100 days to 6 months. The low estimated log BCF value (0.85) indicates that DCE bioconcen trates poorly in aquatic organisms. Since DCE is soluble and adsorbs poorly to the soil and sediments, it can be expected to leach easily into the ground water. Environmental Exposure According to the 1987 TRI data, a total of 6.13 million lbs/yr of DCE was released to atmosphere from 75 facilities. About 3.56 million lbs was released from ponit sources and 2.58 SAL 000066787 3 million lbs was released from nonpoint sources. About 1.47 million lbs/yr were released to water media: 72,775 lbs were directly discharged from 39 facilities to navigable waterways, and 139,428 lbs were indirectly discharged via offsite transfers to 22 POTWs or private waste water treatment facilities. The indirect discharge is based on the assumption of 90% removal of the originally transferred amount (1,394,278 lbs) prior to releases. About 3,173 lbs were directly released to the land and 1.31 million lbs was released to land via underground injection. Using the Industrial Source Complex Long-Term (ISCLT) model in the GEMS Atmospheric Modeling Subsystem (GAMS), EED estimated the air concentrations within a 50 km diameter of all air release sources. Assumptions used in the modeling efforts include 1) 99.9% destruction efficiency for off-site transfers and 2) no decomposition or transformation of DCE in the ambient air after release since the travel time (3 hr) to reach 50 km distance is smaller than the half-life (122 days) of DCE in air. The esti mated air concentrations range from 9.7 x 10'12 ug/m3 to 210 ug/m3 d pending on the specific site release amounts and the distances from the emission sources. EED also estimated surface water and drinking water concen trations downstream from the water release sites using the Routing Graphic Display System. Removal of DCE via absorption and biodegradation in the stream during the travel time was taken into consideration. The ten highest estimated DCE concentrations in surface water are in the range of 13.26 ppb to 798.05 ppb. The highest estimated DCE concentration at a drinking water utility intake is 798 ug/1. Since DCE is expected to leach easily in the ground, the potential for ground water contamination can be high if the land release controls are not effective. Health Aspects Summary Pharmacokinetics DCE is easily absorbed by humans and animals via various exposure routes (inhalation, dermal, ingestion) and rapidly distributed to body tissues. Conjugation of DCE and metabolite 1,2-dichloroethanol with glutathione is the preferred metabolic pathway with S-(2hydroxyethyl)-cysteine and thiodiglycolic acid as the two end metabolic products. Urine excretion is the major route for elimination of DCE and its metabolites. Cases of fatality in humans were reported via inhalation or oral route, in animals, acute inhalation and oral exposure to SAL 000066788 4 DCE in sufficient concentrations result in death. The acute inhalation LC50 in rats is 1,000 ppm (0.1- 8 hr) and oral LD50 rats and mice were 680 and 413 mg/kg/d, respectively. Gross observations at necropsy revealed liver and kidney effects. in Longer term exposure to DCE led to immnunoligical, hematological, hepatic, renal, cardiological and adrenal gland effects in various animals. The inhalation NOAEL and LOAEL for immunological effects (increased susceptibility to bacteria) in mice were 2.5 and 5 ppm (5 d, 3 h/d) which are equivalent to 1.6 and 3.5 mg/kg/d, respectively. The oral LOAEL (via gavage) for immunological effects (reduced antibody forming cell response) in mice was 4.9 mg/kg/d (14 d). The inhalation NOAEL and LOAEL for hematological effects (increased plasma prothrombin clotting time) in monkeys were 100 and 400 ppm (8-12 d, 5 d/wk, 7 h/d, which are equivalent to 33 and 131 mg/kg/d). The oral NOAEL and LOAEL for hematological effects (decrease in leukocytes) in mice were 4.9 and 49 mg/kg/d (14 d). The inhalation NOAEL and LOAEL for hepatic effects in monkey were 100 and 400 ppm (8-12 d, 5 d/wk, 7 hr/d), resepectively. The inhalation LOAELs for liver weight increase and liver degeneration in guinea pigs were 100 and 200 ppm (246 d, 5 d/wk, 7 hr/d, equiv. to 42 and 84 mg/kg/d). The oral NOAEL and LOAEL for hepatic effects in rats were 30 and 80 mg/kg/d (5-7 wk) . The inhalation NOAEL and LOAEL for renal effects in monkeys were 100 ppm and 400 ppm (8-12 d, 5 d/wk, 7 h/d), respectively. The inhalation renal NOAEL in monkeys and rabbits, at longer exposure (25 wk, 5d/wk, 7 h/d) was 200 ppm. In rats, the chronic oral NOAEL for renal effects in rats was 25 mg/kg/d (2 yr). The inhalation LOAEL for cardiological (fatty) and adrenal medulla calcification in monkeys was 200 ppm (25 wk, 5 d/wk, 7 h/d) which is equivalent to 65 mg/kg/d. There were no respiratory effects observed in mice exposed at oral dose of 49 mg/kg/d for 14 d, or 189 mg/kg/d for 90 d, or at air concentration of 100 ppm for 4 wk, 5 d/wk, 7 hr/d (equiv. to 145 mg/kg/d). No respiratory effects were observed in rabbits and monkeys at 200 ppm for 25 wk, 5 d/wk, 7 h/d. No neurological effects were observed in dogs exposed to 400 ppm for 8 months, 5d/wk, 7 h/d (equiv. to 116 mg/kg/d). oooo^89 5 Carcinogenicity EPA classified DCE as a B1 "probable human carcinogen" based on the observation of sufficient evidence of carcinogenicity from animal studies. DCE in corn oil was administered by gavage to rats for 78 weeks at TWA dosages of 47 and 95 mg/kg/d. The incidences of hemangiosarcoma and forestomach squamous-cell carcinomas were significantly increased and were dose-related (NCI, 1978). Based on this study and the assumption of 100% absorption at the low dose, EPA developed oral and inhalation slope factor of 9.1 x 10'2 (mg/kg/d)'1. - Mutagenicity The mutagenic activity of DCE has been tested in bacteria, plants, melanogaster. mammalian cells in vitro and in vivo rodent assays. In gj. typhimurium testing system, DCE has generally elicited marginally positive mutagenesis without metabolic activation and stronger mutagenic responses when exogenous metabolic systems were added. Positive mutagenic responses have been reported for DCE in several assays using melanogaster. These include sex-linked recessive lethal assay (inhalation exposure, 0.07% vapor for 4.6 or 8 hrs and food exposure 50 mM) and somatic cell mutations (larvae food exposure 0.1, 0.5% in food). DCE induced a doserelated increase in mutation at HGPT (hypoxanthine-guanine phosphorilosyl transferase) locus in cultured CHO cells. Doserelated hepatocyte DNA damage was observed in mice treated via various routes (intraperitoneal, oral, inhalation). Developmental and Reproductive Effects No studies were located regarding reproductive effects in humans following oral or inhalation exposure to DCE. No reproductive effects were observed in rats exposed to 150 ppm (1 generation, 7 d/wk, 6 h/d (equiv. to 132 mg/kg/d) or at 25 mg/kg/d (oral administration) for 2 yr. No significantly increased incidence of malformations was observed among the offspring of rats exposed to 100 ppm DCE (NOAEL) during days 6-15 of gestation. Exposure to 300 ppm produced high maternal mortality. Only one dam of six survivors had implantation sites, all of which were resorbed. Administration of 510 mg/kg/d (NOAEL) in drinking water to pregnant mice during days 7-14 of gestation produced no developmental effects and few discernible effects on maternal health; no skeletal or visceral anomalies in the fetus could be attributed to treatment. No dose-dependent effects on fertility, gestation, litter size, viability, or lactation indices were observed in mice exposed to doses of 5-50 (NOAEL) in drinking water for 24-49 weeks. Sal 00() 6 Environmental Effects The 24-, 48-, 72- and 96-hour LC50 values for fathead minnows were 141, 118, 116 and 116 mg/1, respectively. The 7-day LCso in guppies was 106 mg/1. The ECo, ECso, and EC10o were 1.3, 1.8 and 2.4 mg/1 in the golden orfe (duration not specific). In the marine dab, a 96-hr LC50 of 115 mg/1 was reported. The 24- and 48-hr LC50 in fi. maana. have resulted in 24and 48- hour LC50 values of 246 and 218 mg/1 and a NOEC of < 68 mg/1. In the mysid shrimp, 24-, 48-, 72- and 96- hr LCjo values were 108, 110, 112 and 113 mg/1. The 32-day MATC in fathead minnows were 29 and 59 mg/1. The LOELs for reproduction and growth in fi. maana were 21 and 72 mg/1, respectively. In algae, . capricornutum the 7-d toxicity threshold for decreased cell multiplication was 710 mg/1. An EC50 of 130 mg/1 for algae H. pluialis was reported. An average MATC of 41 mg/1 for parental fish weight reduction was identified in a 32-day reproduction bioassAy of DCE using fathead minnows. The NOECs and LOELs for reproduction and growth in maana were 11-42 and 21-72 mg/1, respectively. Based on these two studies, EEB/HERD recommended the chronic concern level of 16 ppm. The OWRS docummented fresh chronic LOEL of 20 ppm. The OWRS also documented LOELs for fresh and marine acute of 118 and 113 ppm, respectively. Preliminary Quantitative Cancer Risk Estimates Ambient Air 6 Drinking Water EED has estimated risk levels and lifetime excess cancer cases due to DCE air emissions. Data used in the estimations included the EPA established cancer risk slope factor of 9.1 x 10'2 (mg/kg/day)model-estimated ambient air concentrations, and population data within GAMS. The estimates are summarized in Table I. The model estimates that there are 65.7 excess life time cancer cases among a total 72 million persons exposed. The annual incidence is 0.939 cases per year. EED has also estimated the lifetime excess cancer risk of populations served by water supplies containing DCE using the model estimates, the EPA established slope factor (oral) of 9.1 x 10'2 (mg/kg/day)' , and census data within RGDS. The ten highest estimates are summarized in Table II. Sfiii 000066791 Table I. 7 Cancer Risk via Air Contamination risk level population at risk life time cases 5.48E-03 .48E-03 - 1.00E-03 .OOE-03 - 1.00E-04 .00E-04 - 1.00E-05 .00E-05 - 1.00E-06 .00E-O6 - 1.00E-07 .00E-07 - 1.00E-08 . 00E-08 - 1.00E-09 .00E-09 -- 1.00E-10 1,165 1,847 50,650 726,412 6,917,031 27,235,465 16,668,806 12,778,101 8,075,259 6.39E+00 2.63E+00 1.09E+01 1.69E+01 1.85E+01 9.59E+00 7.72E-01 4.72E-02 4.35E-03 TOTAL 72,454,736 65.7 Table II. Cancer Risk via Drinking Water Contamination concentration (ug/1) 798.050 0.048 0.046 0.046 0.037 0.029 0.027 0.024 0.024 0.023 risk level 2.07E-3 1.31E-8 1.31E-8 1.31E-8 1.06E-8 8.29E-9 7.71E-9 6.86E-9 6.86E-9 6.57E-9 population at risk 39,800 61,020 8,000 50,000 60,000 3,200 10,488 228,000 600,000 6,000 lifetime cases 8.266E+01 7.623E-03 9.578E-04 5.986E-03 5.778E-03 2.415E-04 7.916E-04 1.424E-02 3.748E-02 3.592E-04 TOTAL 1 ,066,508 82.733 SAL 00006679 SAL 000066793 8 TABLE III highest daily intake via drinking water: highest daily intake via inhalation: 0.022 mg/kg/d 0.06 mg/kg/d oral inhal MOE* NOAEL LOAEL NOAEL LOAEL repro- terato- immuno- heroato- hepatic renal 25 50 4.9 30 25 4.9 49 80 cardio- adrenal neuro65 65 50 1.6 33 32 3.3 131 42 131 116 via DW via inhal H36 416 2272 833 72 27 72 27 1363 500 1136 416 2954 1083 2954 1083 5272 1933 Lowest NOAEL (or LOAEL if no NOAEL available) was used in the mgf r-air. i under each health effect category calculations ** All NOAELs and LOAELs are expressed in rog/kg/d. 9 As indicated in the table II, approximately 40 thousand persons consume drinking water (from one Indiana facility) containing DCE estimate of 798 ppb. This may result in an excess of 82.66 lifetime cancer cases. The lowest NOAELs and LOAELs for various health effects are listed in Table III. The MOEs of the highest daily DCE inhalation intake for various effects are also summarized in Table III. The low MOE values for immunological and hematological effects suggests further testing needs for DCE. Surface Water The ten highest mean flow estimates range from 3.26 to 798.05 ppb. The estimates are lower than the freshwater chronic concern level of 16 ppm recommended by EEB/HERD. EED estimated that the potential of DCE leaching to the ground water from the landfill sites can be high because of the high water solubility and low absorbability of DCE. Recrulatorv Activities DCE has several state and federal environmental regulations and activities including two under SARA Title III (EPCRA, 1986). DCE is listed as a Toxic Release Inventory (TRI) chemical under EPCRA Section 313. Under CERCLA, releases of DCE at or above the reportable quantity (RQ) of 5,000 lb must be reported to the Region or National Response Center. It is also selected for a Toxicological Profile under CERCLA Section 104. DCE is on the RCRA Appendix IX list of chemicals for which ground water at RCRA hazardous waste sites is. required to be.monitored. ------------------ DCE is considered as a toxic pollutant under the Toxic Pollutant Effluent Standard of cwa. In 1980, OWRS recommended LOEL (Lowest Observed Effect Level) of 118 ppm and 20 ppm for acute and chronic freshwater aquatic life. OWRS estimated that ingesting water and organisms containing 0.94 ppb DCE may result in a cancer risk of 1 x 10"6. Consumption of aquatic organisms at 243 ppb concentration may result in a cancer risk of 1 x 10 The Maximum Contaminant Level drinking water standard for DCE is 5 ug/1. The ODW health advisories for 1-day, 10-day, and long term are 0.74, 0.74, and 2.6 ug/1, respectively. Nine states have maximum acceptable concentration standards for drinking water ranging from 0.38 to 5 ppb. Eleven states have maximum acceptable ambient air concentration standards ranging from 0.038 to 1,000 ug/m3. DCE is a pesticide active ingredient under FIFRA. Under OSHA, the Permissible Exposure Limits (PEL) for 8-hr TWA, SAL 00006&794 JMW585 1 SECTION 1. SHORT TITLE AND TABLE OF CONTENTS. 2 This Act may be cited as the ''Pollution Prevention Act 3 of 1990". TABLE OF CONTENTS Sec. 1. Short title and table of contents. Sec. 2. Findings and policy. Sec. 3. Definitions. Sec. 4. EPA activities. Sec. 5. Grants to States for State technical assistance programs. Sec. 6. Source reduction clearinghouse. Sec. 7. Source reduction and recycling data collection. Sec. 8. EPA report. Sec. 9. Savings provisions. Sec. 10. Authorization of appropriations. Sec. 11. Implementation. 4 SEC. 2. FINDINGS AND POLICY. 5 (a) FINDINGS.--The Congress finds that: 6 (1) The United States of America annually produces 7 millions of tons of pollution and spends tens of billions 8 ' of "dollars per year controlling this.pollution. 9 (2) There are significant opportunities for industry 10 to reduce or prevent pollution at the source through 11 cost-effective changes in production, operation, and raw 12 materials use. Such changes offer Industry substantial 13 savings in reduced raw material, pollution control, and 14 liability costs as well as help protect the environment 15 and reduce risks to worker health and safety. 16 (3) The opportunities for source reduction are often SAl. 000066795 JMW585 3 1 not realized b cause existing regulations, and the 2 industrial resources they require for compliance, focus 3 upon treatment and disposal, rather than source 4 reduction; existing regulations do not emphasize 5 multi-media management of pollution; and businesses n ed 6 information and technical assistance to overcome 7 institutional barriers to the adoption of source 8 reduction practices. 9 (4) Source reduction is fundamentally different and 10 more desirable than waste management and pollution 11 control. The Environmental Protection Agency needs to 12 address the historical lack of attention to source 13 reduction. 14 (5) As a first step in preventing pollution through 15 source reduction, the Environmental Protection Agency 16 must establish a source reduction program which collects 17 and disseminates information, provides financial 18 assistance to States, and implements the other activities 19 provided for in this Act. 20 (b) POLICY.--The Congress hereby declares it to be the 21 national policy of the United States that pollution should be 22 prevented or reduced at the source whenever feasible; 23 pollution that cannot be prevented should be recycled in an * 24 environmentally safe manner, whenever feasible; pollution 25 that cannot be prevented or recycled should be treated in an 006O796 00 JMW585 4 1 environmentally safe manner whenever feasible; and disposal 2 or other release into the environment should be employed only 3 as a last resort and should be conducted in an 4 environmentally safe manner. 5 SEC. 3. DEFINITIONS. 6 For purposes of this Act-- 7 (1) The term ''Administrator" means the 8 Administrator of the Environmental Protection Agency. 9 (2) The term "Agency" means the Environmental 10 Protection Agency. 11 (3) The term ''toxic chemical" means any substance 12 on the list described in section 313(c) of the Superfund 13 Amendments and Reauthorization Act of 1986. 14 (4) The term ''release" has the same meaning as 15 provided by section 329(8) of the Superfund Amendments 16 and Reauthorization Act of 1986. 17 - ----- (5) (A)-The term - 'source reduction" means any............... 18 practice which-19 (ij reduces the amount of any hazardous 20 substance, pollutant, or contaminant entering any 21 waste stream or otherwise released into the 22 environment (including fugitive emissions) prior to 23 recycling, treatment, or disposal; and 24 (ii) reduces the hazards to public health and the 25 environment associated with the release of such SAL. 000066797 JHW535 5 1 substances* pollutants* or contaminants. 2 The term includes equipment or technology modifications* 3 process or procedure modifications* reformulation or 4 redesign of products* substitution of raw materials* and 5 improvements in housekeeping* maintenance* training, or 6 inventory control. 7 (B) The term "source reduction" does not include 8 any practice which alters the physical* chemical* or 9 biological characteristics or the volume of a hazardous 10 substance* pollutant* or contaminant through a process or 11 activity which itself is not integral to and necessary 12 for the production of a product or the providing of a 13 service. 14 (6) The term "multi-media" means water* air* and 15 land. 16 (7) The term "SIC codes" refers to the 2-digit code 17 numbers used for classification of economic activity in 18 the Standard Industrial Classification Manual. 19 SEC. 4. EPA ACTIVITIES. 20 (a) AUTHORITIES.--The Administrator shall establish in 21 the Agency an office to carry out the functions of the 22 Administrator under this Act. The office shall be ind pendent 23 of the Agency's single-medium program offices but shall have 24 the authority to review and advise such offices on their 25 activities to promote a multi-media approach to source SAL 000066798 JKT585 6 1 reduction. The office shall be under the directi n of such 2 officer of the Agency as the Administrator shall designate. 3 (b) FUNCTIONS.--'The Administrator shall develop and 4 implement a strategy to promote source reduction. As part of 5 the strategy, the Administrator shall-- 6 (1) establish standard methods of measurement of 7 source reduction; 8 (2) ensure that the Agency considers the effect of 9 its existing and proposed programs on source reduction 10 efforts and shall review regulations of the Agency prior 11 and subsequent to their proposal to determine their 12 effect on source reduction; 13 (3) coordinate source reduction activities in each 14 Agency Office and coordinate with appropriate offices to 15 promote source reduction practices in other Federal 16 agencies, and generic research and development on 17 techniques and processes which have broad applicability; 18 (4) develop improved rethods of coordinating, and' 19 assuring public access to data collected under Federal 20 environmental statutes; 21 (5) facilitate the adoption of source reduction 22 techniques by businesses. This strategy shall include the 23 use of the Source Reduction Clearinghouse and Stat 24 matching grants provided in this Act to foster the 25 exchange of information regarding source reduction SQL 066?99 JMW585 1 techniques, the dissemination of such information to 2 businesses, and the provision of technical assistance to 3 businesses. The strategy shall also consider the 4 capabilities of various businesses to make use of source 5 reduction techniques; 6 (6) identify, where appropriate, measurable goals 7 which reflect the policy of this Act, the tasks necessary 8 to achieve the goals, dates at which the principal tasks 9 are to be accomplished, required resources, 10 organizational responsibilities, and the means by which 11 progress in meeting the goals will be measured; 12 (7) establish an advisory panel of technical experts 13 comprised of representatives from industry, the States, 14 and public interest groups, to advise the Administrator 15 on ways to improve collection and dissemination of data; 16 (8) establish a training program on multimedia source 17 reduction opportunities, including workshops and guidance 18 documents, for State and Federal permit issuance, 19 enforcement, and inspection officials working within all 20 agency program offices. 21 (9) identify and make recommendations to Congress to 22 eliminate barriers to source reduction including the use 23 of incentives and disincentives; 24 (10) identify opportunities to use Federal 25 procurement to encourage source reduction; SAL 000066800 jnnsoD 8 1 (11) develop, test and disseminate model source 2 reduction auditing procedures designed to highlight 3 source reduction opportunities; and 4 (12) establish an annual award program to recognize a 5 company or companies which operate outstanding or 6 innovative source reduction programs. 7 SEC. 5. GRANTS TO STATES FOR STATE TECHNICAL ASSISTANCE 8 PROGRAMS. 9 (a) GENERAL Authority.--The Administrator shall make 10 matching grants to States for programs to promote the use of 11 source reduction techniques by businesses. 12 (b) CRI TER I A. "When evaluating the requests for grants 13 under this section/ the Administrator shall consider, among 14 other things, whether the proposed State program would 15 accomplish the following: 16 (1) Make specific technical assistance available to 17 businesses seeking information about source reduction 18 opportunities, including'funding'for'experts to provide 19 onsite technical advice to business seeking assistance 20 and to assist in the development of source reduction 21 plans. 22 (2) Target assistance to businesses for whom lack of 23 information is an impediment to source reduction, 24 (3) Provide training in source reduction techniques. 25 Such training may be provided through local engineering SAL 0000(66801 JMX503 9 1 schools or any other appropriate means. 2 (c) HATCHtNG Funds*--Federal funds used in any state 3 program under this section shall provide no more than 50 per 4 centum of the funds made available to a State in each year of 5 that State's participation in the program. 6 (d) Effectiveness*--The Administrator shall establish 7 appropriate means for measuring the effectiveness of the 8 State grants made under this section in promoting the use f 9 source reduction techniques by businesses. 10 (e) INFORMATION.--States receiving grants under this 11 section shall make information generated under the grants 12 available to the Administrator. 13 SEC. 6. SOURCE REDUCTION CLEARINGHOUSE. 14 (a) AUTHORITY*--The Administrator shall establish a 15 Source Reduction Clearinghouse to compile information 16 including a computer data base which contains information n 17 managementf technical# and operational approaches to source 13 " r educt ion ."The" Admini s trator shall use" the "Clear inghouse to-- 19 (1) serve as a center for source reduction technol gy 20 transfer; 21 (2) mount active outreach and education programs by 22 the States to further the adoption of source reduction 23 technologies; and 24 (3) collect and compile information reported by 25 States receiving grants under section 5 on the operati n 6b02 0ooo JMW585 10 1 and success of State source reduction programs. 2 (b) Public Availability)--The Administrator shall make 3 available to the public such information on source reduction 4 as is gathered pursuant to this Act and such other p rtin nt 5 information and analysis regarding source reduction as may be 6 available to the Administrator. The data base shall permit 7 entry and retrieval of information to any parson. 8 SEC. 7. SOURCE REDUCTION AND RECYCLING DATA COLLECTION. 9 (a) Reporting Requirements.--Each owner or operat r of a 10 facility required to file an annual toxic chemical release 11 form under section 313 of the Superfund Asandments and 12 Reauthorization Act of 1986 ("SARA") for any toxic chemical 13 shall include with each such annual filing a toxic chemical 14 source reduction and recycling report for the preceeding 15 calendar year. The toxic chemical source reduction and 16 recycling report shall cover each toxic chemical required to 17 be reported in the annual toxic chemical release form filed 18 by the owner or operator under section 313(c) of that Act. 19 This section shall take effect with the annual report filed 20 under section 313 for the first full calendar year beginning 21 after the enactment of this Act. 22 ib) Items Included in Report,--The toxic chemical source 23 reduction and recycling report required under subsection (a) * 24 shall set forth each of the following on a 25 facility-by-facility basis for each toxic chemical: 00066803 SAL 0 wnnvow u 1 (1) The quantity of the chemical entering any waste 2 stream (or otherwise released into the environment) prior 3 to recycling, treatment, or disposal during the calendar 4 year for which the report is filed and the percentage 5 change from the previous year. The quantity reported 6 shall not include any amount reported under paragraph 7 (7). When actual measurements of the quantity of a toxic 8 chemical entering the waste streams are not readily 9 available, reasonable estimates should be made based on 10 best engineering judgment. 11 (2) The amount of the chemical from the facility 12 which is recycled (at the facility or elsewhere) during 13 such calendar year, the percentage change from the 14 previous year, and the process of recycling used. 15 (3) The source reduction practices used with respect 16 to that chemical during such year at the facility. Such 17 practices shall be reported in accordance with the 18.......... following categories unless the Administrator finds other 19 categories to be more appropriate: 20 (A) Equipment, technology, process, or procedure 21 modifications. 22 (B) Reformulation or redesign of products. 23 (C) Substitution of raw materials. 24 (D) Improvement in management, training, 25 inventory control, materials handling, or other SAL 000066804 12 1 general operational phases of industrial facilities. 2 (4) The amount expected to be reported under 3 paragraph (1) and (2) for the two calendar years 4 immediately following the calendar year for which the 5 report is filed. Such amount shall be expressed as a 6 percentage change from the amount reported in paragraphs 7 (1) and (2). 8 (5) A ratio of production in the reporting year to 9 production in the previous year. The ratio should be 10 calculated to most closely reflect all activities 11 involving the toxic chemical. In specific industrial 12 classifications subject to this section, where a 13 feedstock or some variable other than production is the 14 primary influence on waste characteristics or volumes, 15 the report may provide an index based on that primary 16 variable for each toxic chemical. The Administrator is 17 encouraged to develop production indexes to accommodate 18 -------- individual._industri.es._for use on ..a. voluntarybasis._________ 19 (6) The techniques which were used to identify source 20 reduction opportunities. Techniques listed should 21 include, but are not limited to, employee 22 recommendations, external and internal audits, 23 participative team management, and material balance 24 audits. Each type of source reduction listed under 25 paragraph (3) should be associated with the techniques or fc>k' 00 0^ 13 1 multiples of techniques used to identify the source 2 reduction technique. 3 (7) The amount of any toxic chemical released into 4 the environment which resulted from a catastrophic event/ 5 remedial action, or other one-time event and is not 6 associated with production processes during thereporting 7 year. 8 (8) The amount of the chemical from the facility 9 which is treated (at the facility or elsewhere) during 10 such calendar year and the percentage change from the 11 previous year. 12 For the first year of reporting under this subsection, 13 comparison with the previous year is required only to th 14 extent such information is available. is (c) SARA Provisions.--Theprovisions ofsections 322, 16 325(c), and 326 of the Superfund Amendments and 17 Reauthorization Act of 1986 shall apply to the reporting 18 - requirements-of this -section -in- the same manner -as-to-the-- 19 reports required under section 313 of that Act. The 20 Administrator may modify the form required for purposes of 21 reporting information under section 313 of that Act to the 22 extent he deems necessary to include the additional 23 information required under this section. 24 (d) Additional Optional Information,--Any person filing a 25 report under this section for any year may include with the SAL 000066806 JHW585 14 1 report additional information regarding source reduction, 2 recycling, and other pollution control techniques in earlier 3 years. 4 (ej Availability of Data,--subject to section 322 of th 5 Superfund Amendments and Reauthorization Act of 1986, th 6 Administrator shall make data collected under this section 7 publicly available in the same manner as the data collected 8 under section 313 of the Superfund Amendments and 9 Reauthorization Act of 1986. 16 SEC. 8. EPA REPORT. 17 (a) BIENNIAL Reports,--The Administrator shall provide 18 Congress with a report within eighteen months after enactment 19 of this Act an4 biennially thereafter, containing a detailed 20 description of the actions taken to implement the strategy to 21 promote source reduction developed under section 4(b) and of 22 the results of such actions. The report shall include an 23 assessment of the effectiveness of the clearinghouse and 24 grant program established under this Act in promoting the 25 goals of the strategy, and shall evaluate data gaps and data JXW585 15 1 duplication with respect to data collected under Fed ral 2 environmental statutes. 3 (b) Subsequent Reports.--Each biennial report submitted 4 under subsection (a) after the-first report shall contain 5 each of the following: 6 (1) An analysis of the data collected under section 7 7 on an industry-by-industry basis for not less than five 8 SZC codes or other categories as the Administrator deems 9 appropriate. The analysis shall begin with those SIC 10 codes or other categories of facilities which generate 11 the largest quantities of toxic chemical waste. The 12 analysis shall Include an evaluation of trends in source 13 reduction by industry/ firm size/ production/ or other 14 useful means. Each such subsequent report shall cover 15 five SIC codes or other categories which were not c vered 16 in a prior report until all SIC codes or other categories 17 have been covered. 18 (2) An analysis of the usefulness and validity of the 19 data collected under section 7 for measuring trends in 20 source reduction and the adoption of source reduction by 21 business. 22 (3) Identification of regulatory and nonregulatory 23 barriers to source reduction/ and of opportunities for 24 using existing regulatory programs/ and incentives and 25 disincentives to promote and assist source reduction. SAL 000066808 JMW58S 16 1 (4) Identification of industries and pollutants that 2 require priority assistance in multi-media source 3 reduction. 4 (5) Recommendations as to incentives needed t 5 encourage investment and research and development in 6 source reduction. 7 (6) Identification of opportunities and devel pnent 8 of priorities for research and developaent in source 9 reduction methods and techniques. 10 (7) An evaluation of the cost and technical 11 feasibility, by industry and processes, of source 12 reduction opportunities and current activities and an 13 identification of any industries for which there are 14 significant barriers to source reduction with an analysis 15 of the basis of this identification. 16 (8) An evaluation of methods of coordinating, 17 streamlining, and improving public access to data 18 collected under Federal environmental statutes. 19 (9) An evaluation of data gaps and data duplication 20 with respect to data collected under Federal 21 environmental statutes. 22 In thm report following the first biennial report provided 23 for under this subsection, paragraphs (3) through (9) may be 24 included at the discretion of the Administrator. 25 SEC. 9. SAVINGS PROVISIONS. SAL 000066809 \~~ < l vi' (sO~^ DRAFT NTP TECHNICAL REPORT ON THE TOXICITY STUDIES OF 1,2-DICHLOROETHANE IN F344/N RATS, SPRAGUE DAWLEY RATS, OSBORNE-MENDEL RATS, AND B6C3Fi MICE (DRINKING WATER AND GAVAGE STUDIES) D. Morgan, Ph.D., Study Scientist NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park, NC 27709 June 1989 NTP TOX 4 U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service National Institutes of Health These studies were supported in part or whole by funds from the Comprehensive Environmental Response, Compensation, and Liability Act trust fund by interagency agreement with the Agency for Toxic Substances and Disease Registry, U.S. Public Health Service. 0ooo^6i NOT FOR ^1STP:3UTI0N o r a t t r ib u t io n FOREWORD These studies were performed under the direction of the National Institute of Environmental Health Sciences (NIEHS) as a function of the National Toxicology Program (NTP). The NTP, established in 1978, develops and evaluates scientific information about potentially toxic and hazardous chemicals. This knowledge is used for protecting the health of the American people and for the primary preven tion of disease. By bringing together the relevant programs, staff, and resources from the Public Health Service, U.S. Department of Health and Human Services (DHHS), the NTP has centralized and strengthened activities relating to basic and applied research, biological assay development and validation, and the dissemination of toxicological information to the public and to the scientific and regulatory communities. The NTP is made up of four charter DHHS agencies: the National Cancer Institute (NCD, National Institutes of Health; the NIEHS, National Institutes of Health; the National Center for Toxicological Research (NCTR), Food and Drug Administration; and the National Institute for Occupational Safety and Health (NIOSH), Centers for Disease Control. In July 1981, the Carcinogenesis Bioassay Testing Program, NCI, was transferred to the NIEHS. NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTPTOX4, Draft 6/89 OOo6*863 TOXICITY STUDIES 07 1,2-DICHLOROETHANE (ETHYLENE DICHLORIDE) (CAS NO. 107-06-2) IN 7344/N RATS, SPRAGUE DAWLEY RATS, OSBORHE-MENDEL RATS, AND B6C3Fi MICE (DRINKING WATER AND GAVAGE STUDIES) D. Morgan, Ph.D. (Study Scientist) (Tel. No. 919-541-2264) NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park North Carolina 27709 U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service National Institutes of Health 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 SAL 000066863 NOT FOR DISTRIBUTION OR ATTRIBUTION Board Ora:''; CONTRIBUTORS The NTP Report on the Toxicity Studies of 1,2-DichIoroethane is based on the various 13-week stud ies of 1,2-dichloroethane that began in November 1985 and ended in November 1986 at EG&G Mason Research Institute (Worcester, MA). National Toxicology Program (Evaluated Experiment, Interpreted Results, and Reported Findings) D. Morgan, Ph D., Study Scientist John R. Bucher, Ph.D. Michael Elwell, D.V.M., Ph.D. Joel Leininger, D.V.M., Ph.D. B.A. Schwetz, D. V.M., Ph.D. James K. Selkirk, Ph.D. NTP Pathology Working Group (Evaluated Slides and Prepared Pathology Report for Rats and Mice on 7/21/88) John Seely, D.V.M. (Chair) (PATHCO, Inc.) Michael Elwell, D.V.M., Ph.D. (NTP) Jerry Hardisty, D.V.M. (Experimental Pathology Laboratories, Inc.) Margarita McDonald, D.V.M., Ph.D. (NTP) Satoru Motooka, D.V.M. (Eisai Pharmaceutical, Japan) Suzanne Neuenschwander, D.V.M. Experimental Pathology Laboratories, Inc. Brian Short, D.V.M. (Chemical Industry Institute of Toxicology) Principal Contributors at EG&G Mason Research Institute (Conducted Studies and Evaluated Tissues) Herman S. Lilja, Ph.D. A.S. Krishna Murthy, Ph.D. Principal Contributors at Experimental Pathology Laboratories, Inc. (Provided Pathology Quality Assurance) Jerry Hardisty, D.V.M. Suzanne Neuenschwander, D.V.M. Principal Contributors at Analytical Sciences, Inc. (Contractor for Statistical Analysis) Steven Seilkop, M.S. Janet Teague, M.S. Principal Contributors at Carltech Associates, Inc. (Contractor for Technical Report Preparation) William D. Theriault, Ph.D. Abigail C. Jacobs, Ph.D. John Warner, M.S. Naomi Levy, B.A. NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 2 oOOO6606A SA\. un:\ CONTENTS PAGE CONTRIBUTORS .................................................................................................................................................................... ABSTRACT ............................................................................................................................................................................... 2 4 I. INTRODUCTION ........................................................................................................................................................... 7 n. MATERIALS AND METHODS ............................................................................................................................... 23 PROCUREMENT AND CHARACTERIZATION OF U-DICHLOROETHANE ............................... PREPARATION AND CHARACTERIZATION OF DOSE FORMULATIONS IN CORN OIL AND IN DRINKING WATER ........................................................................................................................ THIRTEEN-WEEK STUDY DESIGN ............................................................................................................. STATISTICAL METHODS ............................................................................................................................. DOSE SELECTION ............................................................................................................................................ QUALITY ASSURANCE .............................................................................................. 23 23 28 30 30 30 HI. RESULTS ......................................................................................................................................................................... 31 THIRTEEN-WEEK STUDIES IN RATS ....................................................................................................... DRINKING WATER STUDIES................................................................................................................ GAVAGE STUDIES .................................................................................................................................... 31 31 39 THIRTEEN-WEEK STUDIES IN MICE ........................................................................................................ 41 IV. DISCUSSION AND CONCLUSIONS ...................................................................................................................... 43 V. REFERENCES .............................................................................................................................................................. 51 - APPENDIX APPENDIX A ORGAN WEIGHT. HEMATOLOGIC. AND SERUM CHEMICAL DATA IN THE THIRTEEN-WEEK STUDIES OF U-DICHLOROETHANE ............................... A-l o IDco 5 oc O zo o co u H a oc O O 3 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 SAL 000066865 * Board Dra:"1. HH H--C-- C_H Cl Cl 1,2-Dichloroethane CAS No. 107-06-2 C2H4CI2 Molecular weight 98.97 Synonyms: Ethylene dichloride; 1,2-bichloroethane; a,p-dichloroethane; sy/n-dichloroethane; ethylene chloride; glycol dichloride Trade Names: Freon 150; Brocide; Dutch liquid; Dutch oil ABSTRACT Studies were conducted to compare the toxicity of 1,2-dichloroethane in F344/N rats, Sprague Dawley rats, Osborne-Mendel rats, and B6C3Fi mica. Rats and mice of each sex were exposed to 1,2-dichloroethane in drinking water at 0, 500, 1,000, 2,000, 4,000, or 8,000 ppm for 13 weeks. In addition, F344/N rats of each sex were administered 1,2-dichloroethane in corn oil by gavage to compare toxicity resulting from bolus administration with that of continuous exposure in drinking water. Gavage doses of 1,2dichloroethane were within the range of daily doses resulting from exposure in drinking water. No compound-related deaths occurred in any of the rat strains exposed to 1.2- dichloroethane in drinking water. Weight gain depression was common in each sex of all three rat strains in the 4,000- and 8,000-ppm groups throughout the studies. Water consumption was decreased by 50%-60% with increasing dose for all exposed male and female rats regardless of strain. NOT FOR DISTRIBUTION OR ATTRIBUTION 1.2-Dichloroethane, NTP TOX 4, Draft 6/89 4 ,0 0 c,{A- NOT FOR DISTRIBUTION OR ATTRIBUTION NOT FOR DISTRIBUTION OB a t t r ir i it io m d.-z u/.'i:: Kidney and liver weights were increased in dosed rats of all three strains. No chemical-related lesions were observed except for a dose-related incidence of renal tubular regeneration in female F344/N rats. Nine of 10 female mice exposed to 8,000 ppm 1,2-dichloroethane in drinking water died before the end of the study. Mean body weights of males at 500 ppm or more and females at 1,000 ppm or more were lower than those of controls throughout most of the studies. Kidney weights were' significantly increased for dosed males and females. Renal tubular cell regeneration was seen in males at 8,000 ppm; at 4,000 ppm, minimal regeneration was present in 8/10 male mice. All male F344/N rats that deceived 240 or 480 mg/kg and 9/10 females that received 300 mg/kg 1,2-dichloroethane by gavage died before the end of the studies. Mean body weights of the highest dose males and females were lower than those of vehicle controls throughout the studies. Liver and kidney weights were increased for dosed males and females; however, no compoundrelated lesions were observed. Necrosis of the cerebellum, hyperplasia, inflammation, and mineralisation of the forestomach, and necrosis of the thymus were seen in animals that died or were killed in moribund condition. Rat strain differences in susceptibility to 1,2-dichloroethane toxicity were not apparent at the drinking water concentrations used in these studies; only female F344/M rats exhibited mild chemical-related renal lesions. Male B6C3Fi mice appeared to be more susceptible than rats to toxicity of 1,2dichloroethane administered in drinking water; renal tubule regeneration was 5 1,2-Dichloroethane, NTPTOX4, Draft 6/89 Boars Draft observed in male mice in the 4,000- and 8,000-ppm groups. The higher toxicity in mice was likely due to higher water consumption (milliliters per kilogram body weight), resulting in up to tenfold higher doses to mice than to rats. 1,2-Dichloroethane administered in drinking water resulted in less toxicity to F344/N rats than administration of similar doses (in milligrams per kilogram per day) by gavage. Z o p D ea oe < oe O zo o S9 9 o ac O 1,2-Dichloroethane, NTPTOX4, Draft 6/89 6 HH H-- C -- C -- H II Cl Cl 1,2-Dichloroethane CAS No. 107-06-2 C2H4CI2 Molecular weight 98.97 Synonyms: Ethylene dichloride; 1.2-bichloroethane; a,(3-dichloroethane; sym-dichloroethane; ethylene chloride; glycol dichloride Trade Names: Freon 150; Brocide; Dutch liquid; Dutch oil I. INTRODUCTION Physical and Chemical Properties 1,2-Dichloroethane (ethylene dichloride) is a low molecular weight, chlorinated, aliphatic hydrocarbon. It is a clear, colorless, oily liquid with a chloroform-like odor (Patterson et al., 1976). Other physical and chemical properties are shown in Table 1. Production and Use 1,2-Dichloroethane is produced commercially either by the vapor- or liquidphase reaction of chlorine with ethylene in the presence of 1,2dibromoethane or a metal chloride catalyst or by reaction of ethylene with oxygen and hydrogen chloride in the presence of a copper(II) chloride catalyst (Drury and Hammons, 1979). The annual production of 13 billion 7 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 SAL 000066869 NOT FOR DISTRIBUTION OR ATTRIBUTION TABLE t. SOME CHEMICAL AND PHYSICAL PROPERTIES OF U OICHLOROETHANE (a) Melting point Boiling point Water solubility Log n-octanol/water partition coefficient Relative density Vapor pressure Flash point Flammability limits Conversion factor - 35* C 83* C 8 69 g/liter at 20* C 1.48 1.23 at 20* C 8.53 kPa (64 mm mercury) at 20* C 13*C (closed cup) 0.25-0.64 g/liter, 6%-l6% by volume l ppm in air * 4.05 mg/m1 (at 25* C and 760 mm mercury) (a)IPCS, 1987 pounds (6 billion kg) in 1986 (USITC, 1987) makes 1,2-dichloroethane one of the largest volume synthetic chemicals produced in the United States. World capacity production of 1,2-dichloroethane was estimated to be 51 billion pounds (23 billion kg) in 1980 (Gold, 1980). About 85% of the 1,2-dichloroethane produced in the United States is used in the synthesis of vinyl chloride, and 2%-4% is used in the production of other chemicals, such as 1,1,1-trichloroethane, trichloroethylene, tetrachloroethylene, vinylidine chloride, and ethyleneamines (IARC, 1979). 1,2-Dichloroethane is used as a lead scavenger in gasoline (IARC, 1979); in 1976, about 92 million kg of 1,2-dichloroethane was used in the United States for this purpose. About 0.1% of 1,2-dichloroethane produced in the United States in 1977 was used in fumigants for grain, upholstery, and carpets and as a solvent for metal degreasing (Gold, 1980). Exposure The greatest potential for human exposure to 1,2-dichloroethane occurs in the industrial setting, where an estimated 1.5 million workers could be at NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 8 O risk (Gold, 1980). The primary contact with 1,2-dichloroethane in the workplace results from its use as a solvent. 1,2-Dichloroethane concentrations ranging from 40 to 300 mg/m3 (Cetnarovicz, 1959) have been detected in industrial settings (IPCS, 1987). In a U.S. antiknock-agent blending plant, the maximum exposure concentration measured was 8.9 mg/m3 (Jacobs, 1980). Nonoccupational exposure to 1,2-dichloroethane can occur by inhalation of contaminated air. Singh et al. (1983) estimated the exposure to 1,2dichloroethane from urban air in the United States to be between 8 and 140 pg/day. Near production sites in the United States, an estimated 12.5 million people were exposed to 1,2-dichloroethane at an average annual concentration of up to 40 pg/m3 (Elfers, 1979; Kallam and Dusetzina, 1980). Nonoccupational exposure to 1,2-dichloroethane can also occur by consumption of contaminated water. The National Organics Reconnaissance Survey (Symons et al., 1975) measured 1,2-dichloroethane concentrations of 0-6 pg/liter in finished drinking water in 26 of 80 U.S. cities sampled. Ewing et al. (1977) detected levels of 1,2-dichloroethane greater than 1 pg/liter in surface water from 53 of 204 heavily industrialized U.S. sites. Letkiewicz et al. (1982) estimated that 1,2-dichloroethane levels in all groundwater and surface water systems in the United States are below 10 pg/liter and that most are below 1.0 pg/liter. Daily intake of 1,2-dichloroethane from drinking water containing 10 pg/liter was estimated to be 0.29 pg/kg for a 70-kg adult. 9 1,2-Dichloroethane, NTPTOX4, Draft 6/89 SAL 000066871 NOT FOR DISTRIBUTION OR ATTRIBUTION Symons et al. (1975) observed 1,2-dichloroethane more frequently in finished water than in untreated water, suggesting that contamination may occur during water chlorination (IPCS, 1987). Production of 1,2-dichloroethane by water chlorination has been suggested by others (Versar, 1975; Seufert et al., 1980); however, industrial discharges to surface water and leaching of solid wastes are considered the primary causes of 1,2-dichloroethane contamination in drinking water (Letkiewicz et al., 1982). Absorption and Distribution 1,2-Dichloroethane is rapidly absorbed into the blood of rodents after dermal (Tsuruta, 1975; Jakobson et al., 1982), oral (Sopikov and Gorshunova, 1979; Reitz et al,, 1982), or inhalation (Spreafico et al., 1980; Reitz et al., 1982} exposure. Spreafico et al. (1980) observed that 1,2dichloroethane administered to rats by gavage at doses of 25, 50, or 150 mg/kg was rapidly absorbed, with peak levels in the blood occurring within 20 minutes. Similarly, Reitz et al. (1980, 1982) found that [14C]1,2dichloroethane administered to rats by gavage (150 mg/kg) was completely absorbed. After administration by gavage, 1,2-dichloroethane was found to accumulate most rapidly in the liver, with peak levels attained within 10 minutes of administration (Spreafico et al., 1980). Levels of 1,2-dichloroethane in the lung appeared to be in equilibration with levels in blood. Accumulation in epididymal adipose tissue was slower, with peak levels occurring 45-60 NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 10 Qb b%` minutes after administration; however, these Levels were significantly higher than those in blood. In the same study, Spreafico et al. (1980) compared 1,2-dichloroethane distribution in rats exposed by inhalation (250 ppm for 6 hours) or gavage (50 mg/kg). These doses resulted in comparable peak concentrations of 1,2dichloroethane in blood. After inhalation exposure, peak 1,2-dichloroethane concentrations were higher than after oral exposure in the lung and adipose tissue and lower in the liver. 1,2-Dichloroethane concentrations in the spleen, kidney, and brain were similar to concentrations in blood after administration by either route. During inhalation exposure of rats, equilibrium between blood and tissues (adipose, liver, and lung) was established after 2 hours of exposure at 50 ppm 1,2-dichloroethane and after 3 hours at 250 ppm. In similar studies, Reitx et al. (1980, 1982) investigated the distribution of radioactivity in tissues after oral (150 mg/kg) and inhalation (150 ppm for 6 hours) exposure to [14C]1,2-dichloroethane. During inhalation exposure, equilibration of 1,2-dichloroethane between blood and tissues required 2-3 hours. Target tissues (forestomach, liver, spleen) that developed neoplasms in 1,2-dichloroethane-exposed rats (NCI, 1978), as well as nontarget tissues (kidney, lung, stomach, and remaining carcass homogenate), were surveyed. No striking differences were seen in the distribution of radioactivity in target and nontarget tissues when evaluated 48 hours after oral or inhalation exposure. Levels of radioactivity were U 1,2-Dichloroethane, NTPTOX4, Draft 6/89 SAL 000066873 NOT FOR DISTRIBUTION OR ATTRIBUTION tjodra ora: consistently about two times higher in tissues from animals exposed by gavage than in tissues from animals exposed by inhalation. 1,2-Dichloroethane crosses the placental barrier and has been detected in the fetus. After inhalation exposure of pregnant rats at 1,000 mg/m3 for 4 hours per day, 1,2-dichloroethane was found to accumulate in the placental and fetal tissues over a period of 7 days (Vosovaya, 1977). Withey and Karpinski (1985) also demonstrated that inhalation exposure of pregnant rats resulted in dose-dependent accumulation of 1,2-dichloroethane in the fetus. Urusova (1953) reported that 1,2-dichloroethane accumulated in human breast milk (5.4-6.4 mg/liter) during occupational exposure. Metabolism 1,2-Dichloroethane ham been shown to be metabolized extensively via two principal pathways involving microsomal cytochrome P450 and cytosolic glutathione-5-transferase (GST) with reduced glutathione (GSH) (Figure 1). The cytochrome P450-catalyxed metabolism of 1,2-dichloroethane results in an unstable gem-chlorohydrin intermediate that rapidly eliminates hydrochloric acid to form 2-ehloroacetaldehyde, followed by oxidation to chloroacetic acid or reduction to 2-ehloroethanol (Guengerich et al., 1980; IPCS, 1987). These intermediates may undergo further reaction with GS8 and appear as nontoxic urinary metabolites. The GST-dependent metabolic pathways of 1,2-dichloroethane do not occur to any extent with the other chlorinated ethanes (Anders and Jakobson, 1985) ^ 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 12 v ooo a NOT FOR DISTRIBUTION OR ATTRIBUTION Boar a Dri:': NOT FOR DISTRIBUTION OR ATTRIBUTION Ch2 =CMj ethene * GS5G, HCI GS 5-(2-Eormylmethyl)glutathione Glutathione episulfonium ion A OH - GS V OH S-{2-Hydroxyethyl)glutathione Cellular macromolecular adducts csVSG H/p Y SV" NH, S-Carboxymethyl-c-cysteinylglycine S,S'-Ethene bisglutathione 1 i S,$'-Ethene bis-t-cysteine h2o Ot HO NH, O Acvtyl COA N-Acttyltrantferne S-Carboxymethyl-c-cysteine N-Acetyl-S-carboxymethyl-c-cysteine Thiodlaceticacid (thiodiglyeolic acid) FIGURE 1. PROPOSED PATHWAYS FOR 1,2-DICHLOROETHANE METABOLISM (from IPCS, 1987) 13 1,2- Dichloroethane, NTP TOX 4, Draft 6/89 SAL. 000066875 This pathway involves the direct reaction of 1,2-dichloroethane with GSH to form S-(2-chloroethyl)glutathione, which is nonenzymatically converted to a glutathione episulfonium ion that can undergo several fates (IPCS, 1987), Reaction with water results in the formation of S-(hydroxyethyl)glutathione, and reaction with GSH produces ethene bisglutathione. These reaction products undergo further metabolism to nontoxic urinary metabolites. However, the episulfonium ion is a putative alkylating agent that can also form adducts with protein, RMA, and DNA (Inskeep et al., 1986), This pathway is considered to be the major in vivo route for DMA damage by 1,2dichloroethane (Guengerich et al., 1980; Rannug, 1980; Sundheimer et al., 1982; Inskeep et al., 1986; IPCS, 1987). i ; i < < i , t Excretion NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane is excreted rapidly by rats and mice, regardless of the route of exposure. Approximately 89% or more of 1,2-dichloroethane administered to mice by intraperitoneal injection was excreted within 24 hours (Yllner, 1971) or within 48 hours by mice receiving the chemical orally (Mitoma et al., 1985) and by rats exposed by gavage or inhalation (Reitz et al., 1982; Mitoma et al., 1985). Excretion of 1,2-dichloroethane or its metabolites occurs primarily in exhaled air and in urine in rats and mice exposed by various routes (Davidson et al., 1982; IPCS, 1987). Yllner (1971) found that up to 421 of the 1,2-dichloroethane given to mice by intraperitoneal injection was recovered unchanged in the exhaled air. The percentage of unmetabolized 1,2-dichloroethane exhaled was greater at higher doses than at lower doses, indicating a limited capacity for metabolism. $ 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 14 Similarly/ in rats, 29% of an oral dose of 1,2-dichloroethane (ISO mg/kg) and 1.8% of a lower dose administered by inhalation (ISO ppm for 6 hours) were recovered unchanged in the breath (Reitz et al., 1982). Toxicity in Humans Data on the effects of 1,2-dichloroethane in humans are limited to reports of accidental exposures, and many of these are concerned with mixed chemical exposures. Short-term inhalation exposure to 1,2-dichloroethane at high concentrations initially affects the central nervous system. Signs and symptoms include headache, dizziness, weakness, muscle spasms, cyanosis, hypotonia, vomiting, epigastric pain, and diarrhea. Unconsciousness and death may follow. Irritation and inflammation of the respiratory tract result in symptoms of cough and rales. Bronchial inflammation and respiratory insufficiency due to central nervous system depression may result in cyanosis (Koxik, 1957; Cetnarovicz, 1959; USEPA, 1985; IFCS, 1987). Changes in heart rhythm, probably secondary to cardiac sensitization to catecholamines, were reported (Suveev and Babichenko, 1969). Short-term oral exposure of humans to 1,2-dichloroethane produces effects similar to, but more pronounced than, those after short-term inhalation exposure. In addition, ocular effects such as dilation or constriction of the pupils, impairment of eye reflexes (Weiss, 1957; Troisi and Cavallazzi, 1961), conjunctivitis (Henschick, 1957), and corneal opacity (Weiss, 1957) have been reported after oral exposure to 1,2-dichloroethane. 15 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 000066377 SftL NOT FOR DISTRIBUTION OR ATTRIBUTION 3oarc (Draft Toxicity in Animals The effects of short-term (4-9 months) inhalation exposure to 1,2dichloroethane were investigated in several studies in a number of laboratory animal species (Heppel et al., 1946; Spencer et al., 1951; Hofmann et al., 1971). Of the species studied, rats and mice appear to be the most sensitive to the toxic effects of 1,2-dichloroethane. The noobserved-adverse-effect level for short-term exposure (4-9 months) of rats in three investigations is about 100 ppm (IPCS, 1987). The oral LD50 for 1,2-dichloroethane was 413 (female) or 489 (male) mg/kg body weight in CD*-1 mice (Munson et al., 1982), 680-850 mg/kg in rats (McCollister et al., 1956; Larionov and Kokarovtseva, 1976), and 2,500 mg/kg in dogs (Barsoum and Saad, 1934). Spreafico et al. (1980) investigated the effects of long-term 1,2- dichloroethane inhalation exposure on clinical chemistry indices of Sprague Dawley rats. Three-month-old rats of each sex were exposed at 0, 5, 10, 50, Z o or 250 ppm for 7 hours per day, 5 days per week for 3, 6, or 18 months. The |ne e3g highest exposure concentration was reduced to 150 ppm after several weeks cc <t because of high mortality. An additional group of 14-month-old rats was oc o exposed for 12 months at the same 1,2-dichloroethane concentrations. In the Z o older rats, changes were detected in serum aspartate aminotransferase, serum t- 3 eg alanine aminotransferase, and Y-9lutamyl transpeptidase activity and in serum uric acid, blood urea nitrogen, and serum cholesterol concentrations / 5 after exposure for 12 months. These effects were not observed after the 3- cc o month-old animals were exposed for 3, 6, or 18 months. 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 16 Qb fc#,1s 0o Administration of 1,2-dichlocoethane to rats by gavage, five times per week for 2 weeks at doses of 150 mg/kg or less, had no effect on organ or body weights, histology, clinical chemistry, or hematology (Van Each et al., 1977; Reitz et al., 1982). When rats were administered 30 or 90 mg/kg 1,2dichloroethane by gavage, 5 days per week for 13 weeks, decreased weight gain was observed (Van Esch et al., 1977). Relative kidney weights of rats of each sex and relative brain and liver weights of females receiving 90 mg/kg 1,2-dichloroethane by gavage were increased. Histology and clinical chemistry were normal. Six of six rats died after receiving 300 mg/kg 1,2dichloroethane by gavage for 5 days; fatty degeneration of liver and an increase in liver triglycerides were observed (Van Esch et al., 1977). Alumot et al. (1976) observed increased total liver fat and triglycerides in rats after ingestion of approximately 100 mg/kg 1,2-dichloroethane per day in feed for 7 weeks. In a long-term study, rats were administered feed that had been fumigated with 1,2-dichloroethane, resulting in doses of 0, 11-17, or 23-25 mg/kg per day. After exposure for 2 years, no adverse effects were observed on growth, survival, or serum composition. Immunotoxicity Immunosuppression was observed in rabbits exposed to 1,2-dichloroethane at 100 mg/m3 for 3 hours per day, 6 days per week for 7.5-8 months (Shmuter, 1977). Production of antibodies against typhoid vaccine was reduced by 80% in exposed animals, and a concomitant twofold increase in Forsman sheep erythrocyte antibodies was observed. 17 1,2-Dichloroethane, NTPTOX4, Draft 6/89 NOT FOR DISTRIBUTION OR ATTRIBUTION 3o<arc Dnr. Munson et al. (1982) reported a 30% reduction in leukocyte counts in CD^-1 mice administered 49 mg/kg 1,2-dichloroethane by gavage for 14 days. The number of antibody-forming cells in the spleen was decreased by 25% and 40% in mice receiving 4.9 and 49 mg/kg by gavage, respectively. No effects were observed on cell-mediated immunity in a second group of mice receiving 3, 24, or 189 mg/kg 1,2-dichloroethane in drinking water for 13 weeks. Teratology and Reproductive Toxicology Administration of 1,2-dichloroethane either by inhalation (Rao et al., 1980), in drinking water (Lane et al., 1982), or in formulated diets (Alumot et al., 1976) did not affect fertility, nor did it induce embryotoxic, fetotoxic, or teratogenic effects in several species. Vosovaya (1977) observed a possible adverse effect of 1,2-dichloroethane on reproduction after female rats were exposed to 1,2-dichloroethane by inhalation at 15 mg/m3 for 4 hours per day, 6 days per week for 4 months before mating. During this period, the length of the estrous cycle increased. The rats were then mated and the exposure continued. Total embryonal mortality was increased, and preimplantation losses were about five times greater in exposed rats than in controls. In another study (Vosovaya, 1974), female rats were exposed at 57 10 mg/m3 for 4 hours per day, 6 days per week for 6 or 9 months. The fertility of mated females and the weight of newborn rats were reduced, and perinatal mortality was increased. NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 18 Genetic Toxicology 1,2-Dichloroethane has been shown to be mutagenic in a variety of in vitro tests. It induced DMA damage in Escherichia coll (Brem et al., 1974 ; Rasenkranz, 1977) and gene mutations in Salmonella (McCann et al., 1975; Bignami et al., 1977; Rosenkranz, 1977; Simmon et al., 1977; NTP unpublished data). 1,2-Dichloroethane has also been shown to induce sex-linked recessive lethal mutations in Drosophila (Shakarnis, 1969; King et al., 1979; Kramers and Bissumbhar, 1983) and gene mutations in mammalian lymphoblastoid cells (Crespi et al., 1985). Additional effects observed in mammalian cells in vitro include induction of sister chromatid exchanges and chromosomal aberrations in Chinese hamster ovary cells (NTP unpublished data). Although mutagenic in vitro, 1,2-dichloroethane has demonstrated no genotoxic activity in mammalian cells in vivo, as shown by results from a limited number of studies. Analysis of peripheral blood smears obtained from the 13-week study animals showed no increase in micronucleated erythrocytes (NTP unpublished data), and bone marrow micronucleus studies in mice that received one or two intraperitoneal injections of 1,2dichloroethane were also negative (King et al., 1979; Jenssen and Ramel, 1980). SAL 000066881 19 1,2-DichIoroethane, NTP TOX 4, Draft 6/89 NOT FOR DISTRIBUTION OR ATTRIBUTION 3oard Draft Carcinogenicity The potential carcinogenicity of 1,2-dichloroethane was investigated in a number of studies in which 1,2-dichloroethane was administered to rats and mice by various routes. The results of studies evaluating the carcinogenicity of 1,2-dichloroethane are conflicting. The National Cancer Institute carcinogenesis studies of 1,2-dichloroethane conducted in Osborne-Mendel rats and B6C3Fi mice via gavage in corn oil indicated that 1,2-dichloroethane was carcinogenic and caused neoplasms at multiple sites with metastases (NCI, 1978). However, results of inhalation studies in Sprague Dawley rats and Swiss mice were negative (Maltoni et al., 1980). Attempts to reconcile the results of these two conflicting reports have centered around the purity of the study chemical, species and route differences, contamination of the animal room with known carcinogens, and other technical considerations (Maltoni et al., 1980). Although most confounding factors can be excluded, species and route differences remain the most likely reasons for the contradictory findings. Pharmacokinetic data showing more rapidly attained and sustained levels of 1,2dichloroethane in blood of Osborne-Mendel rats after oral exposure, as opposed to inhalation of 1,2-dichloroethane at comparable doses, correlated with greater DMA alkylation after oral exposure (Reitz et al., 1982). A comparable route-specific genotoxic effect was reported by Storer et al. (1984), who showed significant hepatic DMA damage in mice after short-term oral or intraperitoneal administration but not with comparable inhalation exposure to 1,2-dichloroethane. NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 20 Van Duuren et al. (1979) gave female Swiss mice dermal applications of 42 or 126 mg 1,2-dichloroethane in acetone, three times per week for 440-594 days; an increased incidence of lung papillomas was detected in mice given 126 mg. Another group of female mice received on# application of 1,2-dichloroethane, followed 2 weeks later by application of phorbol myristate acetate in acetone three times per week for 428-576 days. Although 1,2-dichloroethane was found to induce a significant increase in the incidences of benign lung papillomas, it did not initiate skin neoplasms. Klaunig et al. (1986) investigated the effect of 1,2-dichloroethane on the incidences of liver and lung neoplasms in male B6C3Fi mice according to a two-stage initiation/promotion protocol. Mice received 10 mg/liter diethylnitrosamine in drinking water for 4 weeks and then 835 or 2,500 mg/liter 1,2-dichloroethane in drinking water for 52 weeks. Neither the incidences of lung or liver neoplasms nor the number of neoplasms per mouse were affected in mice receiving 1,2-dichloroethane alone or after initiation with diethylnitrosamine. Theiss et al. (1977) conducted a pulmonary tumor bioassay with 1,2dichloroethane administered to A/St mice by intraperitoneal injection. Doses were 20, 40, or 100 mg/kg, three times per week for 24 weeks. The number of lung adenomas per mouse increased with dose; however, the number of adenomas was not significantly greater than that in controls. 21 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 000&6B83 SAl- 0 NOT FOR DISTRIBUTION OR ATTRIBUTION Study Rationale 1.2- Dichloroethane was included in the first group of 24 priority chemicals for toxicologic evaluation by the National Toxicology Program (NTP) as part of an interagency agreement between the NTP and the Agency for Toxic Substances and Disease Registry. Drinking water may be an important source of human exposure to 1,2-dichloroethane because of contamination from industrial discharge and because of leaching from dump sites' into surface water and groundwater. An adequate study of 1,2-dichloroethane toxicity and carcinogenicity using oral, nonbolus (i.e., formulated drinking water mixtures or feed) administration has not been conducted. Conflicting results in earlier studies of 1,2-dichloroethane may have been due to differences in routes of administration and/or rat strains (Hooper et al., 1980). Potential differences in toxicity resulting from bolus or continuous administration were investigated by administering 1,2dichloroethane to F344/N rats by gavage or in drinking water; potential differences in rat strain susceptibility to 1,2-dichloroethane toxicity were investigated in F344/M, Osborne-Mendel, and Sprague Dawley rats administered 1.2- dichloroethane in drinking water. NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 22 SAL 000066884 i II. MATERIALS AMD METHODS Procurement and Characterization of 1,2-Dichloroethane 1,2-Dichloroethane was obtained in one lot from 8.F, Goodrich Chemicals Group (Cleveland, OH). Purity and identity analyses were conducted at Midwest Research Institute (MRI) (Kansas City, MO). MRI reports on the analyses performed in support of the 1,2-dichloroethane studies are on file at the National Institute of Environmental Health Sciences. The study material was identified as 1,2-dichloroethane by infrared, ultraviolet/visible, and nuclear magnetic resonance spectroscopy; the purity was determined to be greater than 99 by elemental analysis, Karl Fischer water analysis, potentiometric titration in methanol with 0.01 N aqueous sodium hydroxide to determine free acid content, and gas chromatography. The stability of the chemical during the toxicology studies was monitored by gas chromatography. No deterioration of the 1,2-dichloroethane was seen over the course of the studies. Preparation and Characterization of Dose Formulations in Corn Oil and in Drinking Water The appropriate amounts of 1,2-dichloroethane and corn oil were mixed (w/v) to give the desired concentrations for the gavage studies. Stability studies of 1,2-dichloroethane in corn oil (approximately 10 mg/ml), using gas chromatography, established that the solutions were stable for at least 23 1,2-DichIoroethane, NTPTOX 4, Draft 6/89 SAL 000066885 NOT FOR DISTRIBUTION OR ATTRIBUTION Board Drar. 3 weeks when stored in the dark at room temperature. Solutions maintained under simulated animal-room conditions (open to air and light for 3 hours) had a chemical loss of approximately 4%. During the studies, dose formulations were stored for no longer than 3 weeks at approximately 4s C in serum vials. Three complete sets of corn oil formulations were analysed over the course of the 13-week studies, and all were within specifications (10% of the target concentration) (Table 2). The analysis of the formulations remaining after dosing was completed gave results that were in reasonable agreement with those from samples taken immediately after mixing, indicating no loss of chemical during dose administration. Tvo referee analyses confirmed the results obtained by the study laboratory. TABLE 2. RESULTS OF ANALYSIS OF CORN OIL FORMULATIONS IN THE THIRTEENWEEK GAVAGE STUDIES OF 1J-OICHLOROETHANE Target Concentration (mg/gj Determined Concentration (a) (mg/g) oZ 3.9 6.5 h- 8.1 13.3 0Q 16.1 OC 26.5 t 32.0 < 52.3 63.5 OC 103.4 O 3.8 0.05 6.5 0.23 7.8 0.19 12.9 0.31 15.6 0.55 25.2 0.43 31.1 1.14 (b) 51.3 0.49 62.3 0.59 (0 103.2 z o (a) Mean standard deviation for three determinations unlesa otherwise specified; for each determination, all samplea analysed in duplicate. (b) Results for two dsterminations 00 (c) Results for s single determination OC (/I Q OC O Ik O 1,2-Dichloroethane, NTP TOX 4. Draft 6/89 24 NOT FOR DISTRIBUTION OR ATTRIBUTION M A T FO P n iC T D im it id m For the drinking water formulations, the appropriate amounts of 1,2dichloroethane and deionized water were mixed (v/v) to give the desired concentrations. Stability studies of 1,2-dichloroethane in water (approximately 5 rag/ml), using gas chromatographic analysis of methylene chloride extracts of the water solutions, established that the solutions were stable for at least 3 weeks in the dark at 59 C in sealed bottles. 1,2-Dichloroethane solutions maintained under simulated animal-room conditions (clear glass drinking water bottles under normal room light) had losses of 1,2-dichloroethane of 13%, 22%, and 27% after 1, 2, and 3 days, respectively. Because of concerns about the stability of dose formulations during the toxicology studies, drinking water formulations were stored in sealed bottles for no longer than 3 weeks and drinking water bottles were changed at the end of each day. Three complete sets of drinking water formulations were analyzed over the course of the 13-week studies. Four of the 16 formulations were out of specifications (varied by more than 10% from the target concentration), with values ranging from -12% to -33% of target (Table 3). Samples that TABLE 3. RESULTS OF ANALYSIS OF DRINKING WATER FORMULATIONS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 14-DICHL0R0ETHANE Tarfet Concentration (ppm) Detartnlnad Concentration (a) (ppm) 500 1,000 2,000 4,000 8,000 lb) 462 897 1,767 3,640 7,190 10 153 338 546 146 (a) Mean standard deviation for the determination of three formulation* unless otherwise specified; for each determination, all analyses performed in triplicate, (b) Four formulations were analyied. 4 c V e c t 25 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 SAL 000066887 3 ra Or if; were out of specifications were restirred and reanalyzed and were then found to be within specifications. Two referee analyses confirmed the results obtained by the study laboratory. The analysis of formulations remaining in the drinking water bottles after 24 hours in the animal cages showed that the concentrations of the formulations had decreased an average of 29% (with values ranging from --13% to --53%) of target concentrations. Fresh drinking water mixtures were placed in the cages at the end of each day; thus, animals were exposed at concentrations ranging between the initial concentration and the concentration found at the end of 24 hours. Thirteen-Week Study Design Groups of 20 male rats and 10 female rats of each strain and 10 mice of each sex were exposed to drinking water containing 0, 500, 1,000, 2,000, 4,000, or 8,000 ppm 1,2-dichloroethane for 13 weeks. Groups of 10 or 20 male F344/N rats were administered 0, 30, 60, 120, 240, or 480 mg/kg 1,2dichloroethane in corn oil by gavage 5 days per week. Groups of 10 female F344/N rats were administered 0, 18, 37, 75, 150, or 300 mg/kg in corn oil by gavage on the same schedule. The male and female F344/R rats, Sprague Dawley rats, Osborne-Mendel rats, and B6C3Fi (C57BL/6M, female X C3H/HeM MTV-, male) mice used in these studies were produced under barrier conditions at Taconic Farms (Sprague Dawley rats), Frederick Cancer Research Facility (B6C3Fi mice and F344/N rats), or CAMM Research Institute (Osborne-Mendel rats). Animals were progeny of defined microflora-associated parents that were transferred fr i NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 26 isolators to barrier-maintained rooms. Animals were shipped to the study laboratory at 4 weeks of age. The rats were quarantined at the study laboratory for 11-14 days and mice for 12-14 days. All animals were placed on study at approximately 6 weeks of age. Hematologic and serum chemical analyses were performed on days 3, 7, 14, and 45 and at the terminal kill on groups of 10 male rats of each strain that received 0, 2,000, 4,000, or 8,000 ppm 1,2-dichloroethane in drinking water and on groups of 10 male F344/N rats that were administered 0, 120, 240, or 480 mg/kg 1,2-dichloroethane in corn oil by gavage. Blood (1.2 ml) was drawn from the tail of each animal and analyzed for erythrocyte and leukocyte counts, hemoglobin, hematocrit, mean cell volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration; a qualitative evaluation of number and morphology of platelets, leukocytes, number of reticulocytes, and erythrocyte morphology was performed. Serum samples were analyzed for sorbitol dehydrogenase, creatine kinase, alanine aminotransferase, alkaline phosphatase, and blood urea nitrogen. Rats used for clinical pathology evaluations ware killed without necropsy, and their tissues were not saved. Animals found moribund and those surviving to the end of the studies were humanely killed. A necropsy was performed on all animals not used in hematologic and serum chemical studies. In some instances, a particular organ was autolyzed or lost; thus, the number of animals from which particular organs or tissues were examined microscopically varies and is not 27 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 Sfi> L 00066(3Q9 NOT FOR DISTRIBUTION OR ATTRIBUTION Boirc JriS\ necessarily equal to the number of animals that were placed on study. Tissues examined are listed in Table 4. Organs and tissues were examined for gross lesions. Tissues were preserved in 10% neutral buffered formalin and routinely processed for preparation of histologic sections for microscopic examination. Tissues and groups examined are listed in Table 4. The liver, right kidney, brain, heart, thymus, lung, and right testis were weighed. Upon completion of the histologic evaluation by the laboratory pathologist, slides, paraffin blocks, and residual wet tissues were sent to the NTP Archives for inventory, slide/block match, and wet tissue audit. The slides, individual animal data records, and pathology tables were sent to an independent pathology laboratory where quality assessment was performed, and the results were reviewed and evaluated by the NTP Pathology Working Group (PWG). The target organs reviewed by the PWG were the forestomach, brain, kidney, and thymus for F344/N rats dosed by gavage and the kidney for all rats strains and B6C3Fi mice receiving formulated drinking water. The final diagnoses represent a consensus of contractor pathologists and the PWG. Details of these review procedures have been described by Maronpot and Boorman (1982) and Boorman et al. (1985). NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, N'TPTOX 4, Draft 6/89 28 TABLE 4. EXPERIMENTAL DESIGN AND MATERIALS AND METHODS IN THE THIRTEEN-WEEK STUDIES OF IJ DICHLOROETHANE Drinking Water Studies Gavage Studies Strain and Species F34VN rata, Osborne-Mendel rata, Sprague Dawley rata. and B6C3FJ mice F34^N rata Study Laboratory EG&G Mason Research Institute EG&G Mason Research Institute Size of Study Groups 10 or 20 males and 10 females of each strain and species 10 or 20 males and 10 females Doses 0. 500,1,000. 2.000, 4,000. or 8,000 ppm 1,2-dichloroethane in drinking water Male-0, 30,60,120. 240, or 480 mg/kg 1,2-dichloroethane in corn oil by gavage; female-0,18,37,75,150, or 300 mg/kg; dose vol-5 ml/kg Method of Animal Distribution Animals distributed to weight classes and then assigned to cages by one table of random numbers and to groupa by another table of random numbers Same as drinking water studies Diet NTH 07 Rat and Mouse Ration (Zeigler Bros., Inc., Gardners. PA): available ad libitum Same aa drinking water studies Animal Room Environment F344/N rata-temp: 68*.72* F: hum: 38%-56%; Sprague Dawley rata-temp: 66*-73* F; hum: 37%*5356; Osborne-Mendel rata-temp: 68*-73" F; hum: 3536-53%; B6C3F[ mice-temp: 68*-77* F; hum: 38%-56%; fluores cent light 12 h/d for sll animals Temp-70*-74* F; hum-2456-64%; fluorescent light 12 h/d Age When Placed on Study 6 wk 6 wk Duration of Dosing 13 wk, dosed until necropsy 5 d/wk for 13 wk, dosed st least 2 consecutive days before necropsy Type and Frequency of Observation Observed 2 x d: weighed initially and 1 X wk thereafter Observed 2 X d; weighed initially and 1 X wk thereafter Necropsy. Histologic Examinations, and Supplemental Studies Necropsy performed on all mica and on all rats not used in Necropsy performed on all rats not used in the serial hematologic the serial hematologic and serum chemical studies: the and serum chemical studies; the following tissues examined following tissues examined histologically for all control histologically for all vehicle control and high dose animals, and high dose animals and for female mice receiving 4,000 males receiving 120 or 240 mg/kg, and female* receiving 150 ppm: adrenal glands, brain, esophagus, eyea (if groesiy mg/kg: adrenal glands, brain, esophagus, eyes (if grossly ab abnormal), gallbladder (mice), groee lesions and tissue normal), gross lesions and tissue masses and regional lymph masses and regional lymph nodes, heart, kidneys, large node*, heart, kidney*, larg* intestine, liver, lungs and mainatem intestine, liver, lungs and mainatem bronchi, mammary bronchi, mammary gland, mandibular and mesenteric lymph gland, mandibular and mesenteric lymph nodes, nasal node*, nasal cavity and turbinate*, ovaries, pancreas, parathy cavity and turbinates, ovaries, pancreas, parathyroids, roids, pharynx (if grossly abnormal), pituitary gland, preputial pharynx (if grossly abnormal), pituitary gland, preputial or clitorai glands (rata), prostate, salivary glands, skin, small or clitorai glands (rats), prostate, salivary glands, skin, inteatine, spinal cord and sciatic nerve (if neurologic signs small intestine, spinal cord and sciatic nerve (if neuro present), spleen, sternebrae or femur or vertebrae including logic signs present), spleen, sternebrae or femur or verte marrow, stomach, teste^epididymis/seminal vesicles, thymus, brae including marrow, stomach, testeVepididymia/ thyroid gland, trachea, urinary bladder, and uterus. Hematolo seminal vesicles, thymus, thyroid gland, trachea, urinary gic and serum chemical analyses performed on groupa oflO male bladder, and uterus. Hematologic and serum chemical rata at d 3,7,14, and 46 and at terminal kill. Organ weights analyses performed on groups of 10 male rata of each obtained at necropsy strain at d 3,7,14, and 45 and at terminal kill. Organ weights obtained at necropsy NOT FOR DISTRIBUTION OR ATTRIBUTION 29 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 SAL 0000668'?! Statistical Methods The analysis of organ weight, hematologic, and serum chemistry data was carried out by using the nonparametric multiple comparison procedures of Dunn (1964) or Shirley (1977). Jonckheere's test (Jonckheere, 1954) was used to evaluate the significance of dose-response trends and to determine whether Dunn's or Shirley's test was more appropriate for pairwise comparisons. The incidences of nonneoplastic lesions were assessed by the Fisher exact test. Dose Selection The solubility of 1,2-dichloroethane in water was the limiting factor in setting the high concentration for drinking water studies. The maximum solubility of 1,2-dichloroethane in water is about 9,000 ppm. Gavage doses were selected to be within the range of doses (in milligrams per kilogram per day) ingested by rats exposed to formulated drinking water. Quality Assurance The studies of 1,2-dichloroethane were performed in compliance with Good Laboratory Practices and regulations (21 CPU 58). The Quality Assurance Unit of EG&G Mason Research Institute performed audits and inspections of protocols, procedures, data, and reports throughout the conduct of the studies. The operations of the Quality Assurance Unit were monitored by the MTP, including a site visit during the period of study performance. NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 30 SAL 0000668V2 III. RESULTS THIRTEEN-WEEK STUDIES IN RATS Drinking Water Studies F344/N Rats: No deaths of F344/N rats occurred during the studies (Table 5). Mean body weights of males exposed at 4,000 ppm or more and of females exposed at 8,000 ppm were lower than those of controls throughout the studies (Figure 2). Water consumption at the higher concentrations was about 60% that by controls. The increase in erythrocyte counts, mild decreases in mean cell volume, and the mild increases in blood urea nitrogen TABLE 3. SURVIVAL. MEAN BODY WEIGHTS. AND WATER CONSUMPTION OF F344/N RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1,2-DICHLOROETHANE Concentration Survival (ppm) (a) Mean Body Weights (grams) Final Weight Water Initial Final Change Relative to Consumption (b) (c) Controls (percent) (d) Estimated Intake Dose (e) MALE 0 500 1,000 2,000 4,000 8,000 FEMALE 0 500 1.000 2.000 4,000 8,000 10/10 10/10 10/10 10/10 10/10 10/10 134 2 133 2 133 2 132 2 134 1 133 2 358 4 359 7 358 5 358 3 329 3 302 4 + 223 3 + 226 6 + 225 5 + 226 3 + 195 3 +168 4 10/10 10/10 10/10 10/10 10/10 10/10 109 2 108 1 108 1 108 2 105 3 106 1 202 2 204 3 207 2 199 t 3 195 1 187 t 2 + 93 2 + 96 2 + 99 t L + 92 1 + 90 3 + 81 2 100 100 100 92 84 101 102 99 97 93 25 24 49 21 86 18 147 15 259 14 515 19 18 58 16 102 14 182 12 320 11 601 lat Number surviving/numbr initially in group (b) Initial group mean body weight standard error of the mean. (c) Mean body weight change of the group standard error of the mean Id) Grams per animal per day; not corrected for spillage. (e) Milligrams per kilogram per day based on the mean of the initial and final body weights NOT FOR DISTRIBUTION OR ATTRIBUTION 31 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 SAL 000066893 Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 400 350 FEMALE RATS 0 PPM O MOW A 1000 PPM 2000 PPM 4000 PPM O 8000 PPM WEEKS ON STUDY 200 i.i..H..hI..i 150100 I 8759 WEEKS ON STUDY 10 12 FIGURE 2. GROWTH CURVES FOR F344/N RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1,2-DICHLOROETHANE 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 32 -sW- Boarc ra:\ in the high dose male rats are all indicative of animal dehydration (Table A3). The decrease in mean cell volume (hemoglobin/erythrocytes) may be related to dehydration resulting in an increase in serum osmolarity, with a subsequent loss of water from and shrinkage of the erythrocytes. The absolute and relative kidney weights and relative liver weights were increased for dosed males and females (Tables 6 and 7). No compound-related TABLE 6. ORGAN WEIGHT DATA FOR MALE RATS IN THE THIRTEEN-WEEK STUDIES OF IJ-DICHLOROETHANE (a) Study/Strain/Organ Dost or Concentration OrlnklB| witar studlea F344/N Control 3ody weight (gram**) * * 383 12.0 Kidney Absolute Relative Liver Abeolute Relative 1.232 48 34 0.1S 15.400 600 42.9 2.17 Spragme Dawi*y Body weight (grams) 449 U.O Kidney Absolute Relative Liver Abeolute Relative 1.871 74 42 0 14 18,480 t 790 41.1 1 03 500 ppm 354 6 9 1.343 * 36 3.8 t 0 08 16.500 340 46.3 * 0.93 446 79 1.943 59 4.4 0.11 20.080 590 *45 0 1.15 1,000 ppn 355 4 5 *1.433 28 * *4 0 0.09 16.980 570 47 7 * 1.37 431 t 7.0 1.954 * 58 M 5 0.08 18.810 4- 570 43.6 0.75 O* borne-Meadei Body weight igrmmii 421 25 3 Kidney Abeolute Relative Liver Abeolute Relative b) 1,500 36 b3 7 t 0.28 b 16,230 + 810 <b i39.2 2 01 Gevege ttedka 477 V3.1 1.600 41 3.4 * 0.09 17,830 * 610 37,4 t 0.85 465 t 17 2 **1,751 V 40 38 0.14 **21.060 840 '*5.4 090 F344/N Body weight (great) Kidney Abeolute Relative Liver Absolut* Relative Vefcldk Com0*04 339 4.6 1,324 29 19 0.08 17.000 440 30.2 t 0 87 30 mg/kg 353 8.7 1.441 26 4 1 0.10 :b) 17.080 r 510 bi 10 S t 0,97 2.000 ppm 355 2.8 **1,523 13 **43 0.04 *17,840 250 **50.2 0.49 432 11.3 1.858 74 43 0.11 20.100 790 *48.5 1.11 433 14.0 1.658 59 3 a 0.09 19.310 800 *44.6 1.24 so *fh( 354 9,0 "1.600 54 "4.5 0.08 18.270 540 5L7 0.92 -1,000 ppa `*327 2.8 1.000 ppm *300 4.3 "1.451 +. 18 "4.4 0.08 16.050 4. 330 49,1 0.79 "1,377 * 22 "4.6 Q.07 14,760 340 *49.2 t 0.65 436 79 414 9.2 2,000 52 *4.6 0 11 19.970 "45.9 490 0 82 2.008 55 "4.9 0.11 19.230 560 "46.3 1.20 393 z 11.8 *380 LI.3 1.613 44 *4.l * o 13 15.190 * 310 38-8 - 1 45 1.507 *4,0 15.900 41.9 68 0 18 800 1.59 120 Kg/k| 341 8.1 **1,653 t 47 4'4 9 0 07 (b) 19.400 860 mt 'b) 57.4 083 :a) 5le*a standard error m ajtlliframs i absolute) or milligram* per gram (relative for group* of lOeaiaula ualeu other**** specified: P value* v*. the control* by Dunn's left t Duao, 1004) or Shirley's us* (Shirley, 19771. <b) Niae animal* wen weighed. P<0 OS **P<0.01 NOT FOR DISTRIBUTION OR ATTRIBUTION 33 1,2-Oichloroethane, NTP TOX 4, Draft 6/89 g^L 000066395 Board Draft TABLE 7. ORGAN WEIGHT DATA FOR FEMALE RATS IN THE THIRTEEN-WEEK STUDIES OF 1.2-DICHLOROETHANE (a) Study/Strain/Organ Dose or Concentration Driitkiaf viur itudJftf F344/N Control ; :94 - 2 4 K.dusy Absolut* ilauQva L;vr Abmiut* R* licit* 739 _ 26 3.9 I 0 13 3.829 134 35.3 - 0 83 Sprafn* D*wl#y Kidaay Absolut* S*l*tiv* L:vr Absolut* Relative ) 271 5.3 1.030 36 3.8 - 0 u 11.140 t 350 41.2 1.07 Of 199 * 2 9 l .000 ppm 213 = 10 1 *914 * IB 4,1 0 07 7 268 179 38.6 - 0 60 "985 4 2 16 0 17 "7 627 - 177 363 L 57 2,000 ppm 196 - 2 4 "945 17 "4 3 0 07 7.278 165 37 2 4. 0.75 283 t 7,8 287 c 6.4 271 t 4 3 1.160 27 M l 0.09 11.890 330 42 0 1.49 *1.221 28 4.3 0.13 12.200 880 42.7 * 2.60 *1.211 * 33 "4 3 0 11 10,990 310 40.6 l 32 4.000 ppm 193 l 3 "932 *. 15 "4 8 0 09 7,551 171 "392 * 094 263 * '6.6 *1.208 50 4,6 0.16 11.500 370 43 5 1 37 8.000 ppa .85 - 2.3 "923 15 "50 0 04 7,134 t 147 "385 - 0 81 256 t 4 6 "1,342 t 18 "5 2 t 0.10 ib) 11.950 450 (b) 46.6 t l 41 ) 274 9 9 279 36 271 t 4,7 Kidnsy Absolut* R* Licit* Liver Abeolut* RilA&Tt 394 26 3.3 O.U 10.390 430 37.9 1.04 **1.017 t 13 `3.7 0 08 11.380 380 41.3 0.98 1.041 * 22 "3.9 * 0.08 10.810 230 40.0 0.81 G*v*fe itadl** F1UM Vihkls Control 18 mf/kf 37 236 6.5 *1,020 24 *4.0 t 0.16 10,390 t 430 41.0 2.39 75 270 6.6 "1.098 37 **4 1 0.14 L0.75O 300 39.6 0.73 266 t 11.2 "1.094 33 "4.3 t 0.28 10,100 t 410 .38.6 2.49 150 mf/kf Body weifngr*m)) 190 1.9 190 2.3 194 3.3 197 2.7 192 1.9 Kidney Absolut* RsietiTe Livsr Absolut* RjUuv* 900 * 16 4.2 0.08 7.343 L20 138 7 0.34 717 70 3.8 0.37 9.000 201 12,1 0.87 798 20 4.1 0.09 '7.920 191 40 6 0.61 998 23 *4.6 008 ">.577 197 *43.6 0.69 984 = 9 `3.1 0.08 "9.773 151 "51.0 1.08 a) Msaa standard srror ia aiUlijritni i abeolut*) or nmhframi p*r jmm i relative) for freup* of 10 animait unless otherwise specified; P veluat ts. the controls oy Dunn's test (Dunn. 1904) or Shirley's t*st < Shinty, L977). bi Nins animal* won wetghtd. *P<0.O5 "P<001 clinical signs war# obsarvad. Ranal tubular raganaration was observed in all dosed and control mala rats and consisted of one or more foci of basophilic-staining tubules lined by closely packed tubular epithelium in the cortex or outer medulla of the kidney. The lesion was minimal to mild and occurred in 9/10 rats in each group. Mo difference in severity was sean between groups. The incidence of renal tubular regeneration in females, NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, XTPTOX 4, Draft 6/89 34 aV> however, was dose related and was observed in 9/10 at 8,000 ppm, 3/10 at 4,000 ppm, 2/10 St 2,000 ppm, i/10 at 1,000 ppm, 0/10 at 500 ppm, and in 0/10 controls. This lesion was of minimal severity in all affected rats. Ho lesions attributable to 1,2-dichloroethane were observed in the liver. Sprague Dawley Rats: All Sprague Dawley rats lived to the end of the studies (Table 8). Mean body weights of males and females exposed at 4,000 ppm or more were lower than those of controls throughout the studies (Figure 3). Water consumption by the three highest dosed groups was about half that by controls for males and was less than half that by controls for females. Mild increases in erythrocyte counts, hemoglobin, hematocrit, and blood urea nitrogen at days 3 and 7 in dosed male rats are evidence of mild animal TABLE 8. SURVIVAL, MEAN BODY WEIGHTS, AND WATER CONSUMPTION Or SPRAGUE DAWLEY RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1J-DICHLOROETHANE Concentration Survival (ppm) (a) Mean Body Weights (grams) Final Weight Water Initial Final Change Relative to Consumption (b) (c) Controls (percent) (d) Estimated Intake Dose (e) MALE 0 500 1.000 2.000 4.000 8,000 FEMALE 0 500 1.000 2.000 4.000 8.000 10/10 10/10 10/10 10/10 10/10 10/10 170 2 169 2 169 2 169 2 168 2 169 3 457 a 452 + 7 439 6 436 12 440 8 418 3 + 288 + 10 + 283 7 + 270 6 + 267 12 + 272 7 + 248 7 10/10 10/10 10/10 10/10 10/10 10/10 139 2 144 2 143 2 143 2 141 2 135 2 281 291 + 290 276 270 257 6 8 5 5 7 5 + 141 t + 147 + 147 + 133 + 128 + 123 t 5 3 4 4 6 4 99 96 95 96 91 104 103 98 96 91 43 37 60 30 99 25 165 21 276 19 518 44 33 76 23 106 18 172 16 311 13 531 (a) Number surviving/number initially in group lb) Initial group mean body weight standard error of the mean I c) Mean body weight change of the group t standard error of the mean id) Grama per animal per day; not corrected for spillage. (e) Milligrams per kilogram per day baaed on the mean of the initial and final body weight* NOT FOR DISTRIBUTION OR ATTRIBUTION 35 1,2-Dichlorodthane, NTPTOX 4, Draft 6/89 0000668^7 SAL Board Draft mean body weight in grams 500 130....... FEMALE RATS O 500 ppm A 1000 ppm 2000 ppm # 4000 ppm O 9000 ppm mean body weight in grams 300 .1..!..S ! i250 ,....|....j W ----- A V O > <? >: j g a ! * !! t > 1CC O> f2 150- ...... ,....... a .......' O <> : H 3 too --------- i--------- i --------- 1--------- i--------- i--------- 1--------- 1--------- i--------- CO WEEKS ON STUDY F- a oat FIGURE 3. GROWTH CURVES FOR SPRAGUE DAWLEY RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1,2-DICHLOROETHANE 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 36 00 $&v- Board Draft dehydration (Table A6). The absolute and relative kidney weights for dosed females, relative kidney weights for dosed males, and the relative liver weights for dosed males and females were significantly increased (see Tables 6 and 7). No compound-related clinical signs were observed. Tubular regeneration occurred in the kidney of males and females in all dosed and control groups; the severity and incidence did not differ between groups. No lesions in the liver were attributed to 1,2-dichloroethane administration. Osborne-Mendel Rats.* No compound-related deaths occurred in Osborne-Mendel rats (Table 9). Mean body weights of males exposed at 2,000 ppm or more and of females exposed at 1,000 ppm or more were lower than those of controls throughout the studies (Figure 4). water consumption by the three highest TABLE 9. SURVIVAL. MEAN BODY WEIGHTS, AND WATER CONSUMPTION OF OSBORNE-MENDEL RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF U-DICHLOROETHANE Concentration Survival (ppm) (a) Mean Body Weights (grams) Final Weight Water Initial Final Change Relative to Consumption (b) (c) Controls (percent) (d) Estimated Intake Dose (e> MALE 0 500 1.000 2.000 4.000 8.000 FEMALE 0 500 1.000 2,000 4.000 8,000 (f) 9/10 10/10 10/10 10/10 10/10 L0/10 172 3 171 4 170 3 169 3 172 3 171 3 452 + 15 482 13 468 17 435 14 399 12 382 11 + 281 16 + 311 14 + 298 18 + 266 14 + 227 14 + 211 12 10/10 10/10 10/10 10/10 10/10 10/10 138 3 139 3 138 3 137 3 136 2 138 2 278 12 277 6 275 5 261 4 275 7 258 5 + 140 12 + 137 5 + 138 t 3 + 124 t 3 + 139 5 + 121 4 107 104 96 88 85 100 99 94 99 93 42 35 54 28 88 22 146 19 266 17 492 43 34 32 26 126 23 213 22 428 18 727 (at Number surviving/number imtiaiJy in group i b) Initial group mean body weight standard error of the mean. Subsequent calculations are based on animals surviving to the end of the study. (c) Mean body weight change of the survivors standard error of the mean (d) Grams per animal per day; not corrected for spillage. (e) Milligrams per kilogram per day based on the mean of the initial and final body weights (f) Week of death; 7 NOT FOR DISTRIBUTION OR ATTRIBUTION 37 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 SAL 000066899 Board Draft O MEAN BODY WEIGHT IN GRAMS 500 *30400350300 250 200- MALE RATS Control O 300 ppm A 1000 ppm Q 2000 ppm 4000 ppm O 8000 ppm o "O' O A A o A Q A A,. 0" o . <? o 9 A 0 A a. a 0 4at O A a. - , A... ? 6 4 'o A' s 6* A 150- 100- 6789 WEEKS ON STUDY --r- 10 11 12 --I-- 13 14 500 450 400350- FEMALE RATS Centre! O 300 ppm A 1000 ppm 2000 ppm 4000 ppm O 8000 ppm MEAN BODY WEIGHT IN GRAMS 300- i 250 i..s i..I.*..? i Q .9 5 1! 1 200- i I0oc 150- 3 100CQ b1/1 6 7 89 WEEKS ON STUDY 10 12 13 14 oae FIGURE 4. GROWTH CURVES FOR OSBORNE-MENDEL RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1,2-DICHLOROETHANE 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 38 .o dosed groups was half or Less than half that by controls. The increases in erythrocyte counts, hematocrit, and hemoglobin (day 3) and the decrease in mean cell volume in dosed male rats are evidence of animal dehydration (Table A9). The absolute and relative kidney weights were increased for dosed females, and the relative liver weights were increased for males receiving 1,000 or 2,000 ppm (see Tables 6 and 7). No compound-related clinical signs were observed. Renal tubular regeneration was seen in all dosed and control groups of each sex. Although the incidences were increased in rats administered the higher doses of 1,2-dichloroethane, the increases were not clearly dose related and the severity was not different between groups. Gavage Studies All male F344/N rats that received 240 or 480 mg/kg and 9/10 females that received 300 mg/kg died before the end of the studies (Table 10). Mean body weights of males at 480 mg/kg and of females at 300 mg/kg were lower than those of vehicle controls throughout the studies (Figure 5), The mean body weight for one cage of female vehicle controls was decreased at week 9, possibly due to not receiving water. Compound-related clinical signs included tremors, salivation, emaciation, abnormal postures, ruffled fur, and dyspnea in males at 240 mg/kg and in females at 300 mg/kg. The absolute and relative kidney and liver weights were increased for dosed males and females (see Tables 6 and 7). Hyperplasia, inflammation, and mineralization were seen in the mucosa of the forestomach in animals that died or were killed in a moribund condition (Table 11). Foci of epithelial necrosis were 39 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft TABLE 10, SURVIVAL AND MEAN BODY WEIGHTS OF F344/N RATS IN THE THIRTEEN-WEEK GAVAGE STUDIES OF IJ-DICHLOROETHANE Dose (mg/kg) Survival (a) Mean Bodv Weights i grams) Initial Ib) Final Change (c) Final Weight Relative to Vehicle Controls 'percent) MALE 0 30 60 120 240 480 FEMALE 0 18 37 75 150 300 10/10 10/10 10/10 10/10 Id) 0/10 if) 0/10 10/10 10/10 10/10 10/10 10/10 (g) 1/10 118 4 119 + 5 120 4 120 4 118 4 117 4 104 2 102 2 102 2 104 2 104 + 2 101 2 333 + 4 346 5 349 9 338 + 9 (fl) (e) 193 2 193 2 197 3 199 3 194 3 177 + 215 6 + 226 4 + 229 + 9 + 218 7 (e) <e> + 89 3 + 91 2 +95 + 3 + 95 2 + 90 + 3 + 76 104 105 102 (e) (e) 100 102 103 101 92 (a> Number surviving/number initially in group ib) Initial group mean body weight standard error of the mean. Subsequent calculations are based on animals surviving to the end of the study. (c) Mean body weight change of the survivors standard error of the mean id) Week of death: U.5,5.6.7,8.8,9.l 1 le) No data are reported due to 100% mortality in this group. lf) Weekofdeath: all l <g) Week of death: 1,1,2,2,2,3,5.11.13* * ** TABLE 11. NUMBERS OF F344/N RATS WITH SELECTED LESIONS IN THE THIRTEEN-WEEK GAVAGE STUDIES OF M-DICHLOROETHANE (a) Site/Lesion Group MALE Forestomach Hyperplasia Mineralization Inflammation Cerebellum Necrosis Thymus Necrosis FEMALE Forestomach Hyperplasia Mineralization Inflammation Cerebellum Necrosis Thymus Necrosis Vehicle Control 0 0 0 0 0 Vehicle Control 0 0 0 0 0 120 mg/kg 1 0 1 0 0 75 mg/kg - - 240 mg/kg 5 3 5 3 4 150 mg/kg 0 0 0 0 0 480 mg/kg 2 2 3 0 to 300 mg/kg 3 l 1 3 5 < a) Ten animals were examined microscopically in each group. *P<0.05 vs. vehicle controls **P <0.01 vs. vehicle controls NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 40 SAL 000066902 MEAN BODY WEIGHT IN GRAMS 350 MALE RATS :--3-- ' ' : '6 \8 S Control O 30 mg/kg 300 --H & 61 r*g/*g .................................................................... * A i2O O -3................................... 122 mg/kfl Aa 243 mg/kg O 467 m^/kf aa 1 . :.........- 1- :.....................* *....................................!................ 250 ; .............. a : 4* r; a : 200 H 1* 150 H............i .. t .. ................-........ `............:................................................... 1.......... :........ .. 100'- - --j - T-1 " 1--" --i------- - i i--- i - - i i " i" 8 7 8 9 10 WEEKS ON STUDY " i-------- r -- ---------- i---------- 13 14 350 300H 250 H FEMALE RATS VoM1o Control O IS mgA A 37 mg/tg 74 mg/Vg 148 mg/V 0 294 og/lg MEAN BODY WEIGHT IN GRAMS 200 H e i j i i..i..f' 0 150 H !O oz fi-- ioo- <5 --------- 1---------- 1---------- 1---------- 1---------- 1---------- p---------- 1---------- 1---------- 1---------- 1---------- 1---------- 1---------- r CO 01 2345878910 1112 13 14 15 s WEEKS ON STUDY H JA 5 cfi FIGURE 5. GROWTH CURVES FOR F344/N RATS ADMINISTERED Ou. 1,2-DICHLOROETHANE IN CORN OIL BY GAVAGE FOR THIRTEEN WEEKS Hoz 41 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 SAL 000066903 Board Draft sometimes seen with hyperplasia and inflammation. Necrosis of the cerebellum and of the thymus was also observed. Necrosis in the cerebellum was mainly in the granular layer of the lateral folia, and mineralization was also present in the areas of necrosis in a few animals. Renal tubular regeneration in vehicle control and dosed groups of males or females did not differ in incidence or severity. THIRTEEN-WEEK STUDIES IN MICE Drinking Water Studies: Mine of 10 female mice exposed at 8,000 ppm died before the end of the studies (Table 12). Mean body weights of males exposed at 500 ppm or more and of females exposed at 1,000 ppm or more were lower than those of controls throughout most of the studies (figure 6). TABLE 12. SURVIVAL. MEAN BODY WEIGHTS, AND WATER CONSUMPTION Or MICE IN THE THIRTEEN-WEEK DRINKING WATER STUDIES Of U-DICHLOROETHANE Concentration Survival (ppm) (a) Mean Body Weights fgrams) Final Weight Water Initial Final Change Relative to Consumption (b) (c) Control* (percent) (d) Estimated Intake Dose (e) MALE 0 500 1.000 2.000 4,000 8,000 FEMALE 0 500 1,000 2,000 4,000 8,000 10/10 10/10 10/10 10/10 10/10 10/10 21.2 0.2 20.5 0.4 21.1 0.4 20.8 0.4 20.3 0.2 20.5 0.3 31.4 0.6 28.9 0.6 29.3 0.5 29.4 0.8 28.6 0.7 25.9 0.7 + 10.2 0.4 + 8.4 0.4 + 8.2 0.6 + 8.6 0.7 + 8.3 0.6 + 5.4 0.8 10/10 10/10 10/10 10/10 10/10 If) 1/10 17.1 0.2 17.8 0.3 16.9 0.2 16.9 0.3 17.1 0.3 17.2 0.4 25.9 0.6 24.7 0.5 23.2 0.6 23.7 0.5 23.8 0.6 23.4 + 8.8 0.5 + 6.9 0.4 + 6.3 0.5 +6.8 0.4 + 6.7 0.5 + 4.7 92.0 93.3 93.6 91.1 82.5 95.4 89.6 91.5 91.9 90.3 13.1 12.3 249 11.3 448 9.8 781 16.6 2,710 12.2 4,207 8.1 10.4 244 13.0 647 12.0 1,182 12.7 2,478 12.5 4.926 la) Number surviving/number initially in group ib) Initial group mean body weight standard error of the mean. Subsequent calculations are based on animals surviving to the end of the study. (c) Mean body weight change of the survivors standard error of the mean id) Grams per animal per day: average of determinations from week 2 to week 13: not corrected for spillage. ^ let Milligrams per kilogram per day based on the mean of the initial and final body weight* if) Week of death: 1,1,5,5,9.10.10.11,13 opr 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 42 NOT FOR DISTRIBUTION OR ATTRIBUTION * Board Draft NOT FOR DISTRIBUTION OR ATTRIBUTION 35 3 so < oc o I-- X o C 25-) >O o CD H FEMALE MICE Contra 0 500 ppm A >000 ppm 2000 ppm 4000 ppm 0 8000 ppm iI a ..I.. $ 9 "r o f...f kI i8 8*. 9o 15- 6 7 8 9 10 13 u WEEKS ON STUDY FIGURE 8. GROWTH CURVES FOR MICE IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1,2-DICHLOROETHANE 43 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 SAL. 000066905 Board Draft Water consumption varied greatly from week to week, but overall water consumption by dosed and control groups appeared to be similar. The absolute and relative kidney and liver weights were significantly increased for dosed males and females (Table 13). No compound-related clinical signs were observed. Compound-related lesions were seen in the kidney of male mice and were most prominent at the highest concentration (Table 14). At 8,000 ppm, a minimal-to-moderate tubular cell regeneration consisting of foci of basophilic-staining tubular epithelium was seen in the cortex of the kidney. Karyomegaly in the tubular epithelium, particularly in areas of regeneration, was characterized by nuclei that were slightly enlarged and TABLE 13. ORGAN WEIGHT DATA FOR MICE IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1.2-DICHLOROETHANE (a) Organ Control 500 ppm 1,000 ppm 2.000 ppm 4,000 ppm 8,000 ppm MALE Number weighed 10 Body weight (grams) 30.0 0.73 9 28.0 0.81 10 28.4 0.47 10 29.0 0.79 9 10 28.3 0.68 "25.4 0.65 Kidney Absolute Relative Liver Absolute Relative FEMALE 305 7 10.2 0.22 301 8 *323 7 **358 8 **385 + 9 **379 12 10.8 0.12 11.4 + 0.12 *12.4 0.33 **13,8 0.40 "15.0 0.54 1.455 55 1.490 42 1.519 55 1,571 56 *1.628 54 *1,598 78 48.5 1.06 **53.6 0.91 **53.4 t 1.18 **54.3 1.46 **57.6 1.10 "62.8 2.13 Number weighed 10 8 10 9 to (b) 1 Body weight (grains) 24.0 0.59 23.7 0.52 22.5 t 0.54 22.8 0.57 23.2 0.57 23.0 Kidney Absolute Relative Liver Absolute Relative 191 4 **225 6 **211 5 *212 7 **215 7 8.0 0.23 **9.4 0,21 **9.4 0.17 *9.3 0.24 **9.3 0.22 1.258 39 1,258 52 1.263 34 1.314 56 *1,383 29 52.5 t 0.85 51.5 0.95 *56.0 0.67 *56.1 1.18 **59.7 1.01 217 9.4 1.391 60.5 (a) Mean standard error in milligrams (absolutelor milligrams per gram (p the controls by Dunn's test (Dunn, 1964) or Shirley's test (Shirley, 1977). (b) Not included in statistical analysis *P<0,05 "PcO.Ol /e) unless otherwise specified; P values vs. 1,2-Dichloroethane, NTPTOX4, Draft 6/89 44 cr <o& NOT FOR DISTRIBUTION OR ATTRIBUTION TABLE 14. NUMBERS OF MICE WITH RENAL LESIONS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1.2-DICHLOR0ETHANE (a) Lesion Control 300 ppm 1.000 ppm 2,000 ppm 4.000 ppm MALE Tubular regeneration 0 I 2 2 "8 Karyomegaly 0000 0 Dilatation 0000 0 Protein casta 00 0 0 0 Mineralization 00 0 0 0 FEMALE Tubular regeneration 0 0 0 0 I (a) Ten mice were examined microacopically in each group, *P<0.05 vs.control* **P<0.01 vs. control* 8,000 ppm **9 *I0 *5 *8 5 0 more variable in size chan in controls. Protein casts were present in the lumen of a few tubules and were sometimes associated with tubular dilatation. In addition, foci of mineralization were present in the renal papilla at the highest dose. At 4,000 ppm, minimal tubular cell regeneration was present in 8/10 male mice; a similar change was present in only one or two mice per group at the lower doses. 1 i t ( NOT FOR DISTRIBUTION OR ATTRIBUTION IV. DISCUSSION AND CONCLUSIONS 1,2-Dichloroethane administered at up to 8,000 ppm in drinking water for 13 weeks appeared to be relatively nontoaic for F344/N, Sprague Davley, and Osborne-Mendel rats. No deaths occurred in exposed rats, and body weight changes were similar for all three rat strains of each sex. The high dos level of 1,2-dichloroethane (8,000 ppm) was selected based on limitations in the solubility and palatability of the chemical in drinking water. The maximum solubility of 1,2-dichloroethane in water is about 9,000 ppm (Torkelson and Rowe, 1981). 45 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 SAL 000066907 Weight gain depression was common in males and females in the two higher dosed groups throughout the studies and was likely caused by dehydration due to poor palatability of the formulated drinking water. Water consumption decreased substantially with increasing dose for all exposed male and female rats, regardless of strain. The decrease in water intake, which was as much as 604 at the highest dose level in male and female Qsborne-Mendel rats, indicates that the dose received by all exposed animals was less than the target dose; however, because water intake was reduced at most exposure levels, equivalent exposure did not occur at different dose levels within a strain. The estimated daily intake of 1,2-dichloroethane was similar for each rat strain at each dose level. Rats administered drinking water containing 8,000 ppm 1,2-dichloroethane received an estimated intake of about 500-725 mg/kg per day. This estimated daily intake is close to the reported oral LD50 for 1,2-dichloroethane administered by gavage (680-850 mg/kg) (McCollister et al., 1956); however, intake of this dose over 24 hours rather than as a bolus resulted in little toxicity. 1,2-Dichloroethane toxicity administered by gavage or in formulated drinking water was compared in F344/M rats. Gavage doses were calculated to be approximately equivalent (in milligrams per kilogram) to the range of exposures resulting from the formulated water mixtures. The F344/N rats were more sensitive to 1,2-dichloroethane administered by gavage than in drinking water, as evidenced by the fact that all males receiving 240 and NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTPTOX4, Draft 6/89 46 SAL. 000066908 480 mg/kg and 9/10 females receiving 300 mg/kg died before the end of the s tud ies. Necrosis of the cerebellum, observed in the brains of three males receiving 240 mg/kg and three females receiving 300 mg/kg, appeared to be related to 1,2-dichloroethane administration. Morphologic alterations in cells of the cerebellum, parenchymous changes in the brain and spinal cord, and hyperemia and hemorrhage of the brain have been observed in humans who' died of acute oral poisoning by 1,2-dichloroethane (Hueper and Smith, 1935; Lochhead and Close, 1951). Hyperplasia, inflammation, and mineralisation of the forestomach were observed in eight male and three female F344/W rats dosed by gavage which died or were killed in a moribund condition. Although forestomach lesions were chemical related, they were not considered life threatening. However, hyperplasia of the forestomach epithelium after 13 weeks of exposure may be of significance, since long-term administration of 1,2-dichloroethane by gavage has been shown to cause neoplasms of the forestomach in OsborneMendel rats (NCI, 1978). Thymic necrosis in four mid dose and all high dose males and in five high dose females was attributed to stress in animals that died or were killed in a moribund condition. Administration of bolus doses of 1,2-dichloroethane by gavage may result in saturation of 1,2-dichloroethane elimination and increased levels of 47 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 SAL 000066909 i NOT FOR DISTRIBUTION OR ATTRIBUTION 3oara Draf1. 1,2-dichloroethane in the blood (Reitz et al., 1982). Exposure at lower concentrations of 1,2-dichloroethane over the course of the day (in drinking water or by inhalation) would result in lower peak blood levels and a lower area under the curve (the integral of the 1,2-dichloroethane concentration in blood as a function of time) and the chemical could be rapidly eliminated, even when the total daily dose was equal to the amount administered by gavage (Reitz et al., 1982). This mechanism may explain the greater toxicity for P344/N rats of 1,2-dichloroethane administered by gavage compared with that after drinking water exposure. Based on the significant organ weight changes in rats receiving the chemical by either the drinking water or gavage routes, liver and kidney appear to be target organs for 1,2-dichloroethane. Liver weights were usually increased in rats of all strain, sex, and dose level combinations. The kidney was also increased in weight and was significantly increased more frequently than the liver. Despite increases of 10%-20% in kidney and liver weights, no histologic changes could be clearly attributed to 1,2-dichloroethane, except perhaps for renal tubular epithelium regeneration in female F344/N rats. Serum chemistry data were not indicative of liver or kidney injury. Increased blood urea nitrogen was attributed to dehydration. Regenerative lesions of the rat kidney are commonly seen and are associated with chronic progressive nephropathy, which occurs in most strains of albino rats. The incidence and severity of progressive nephropathy are sex dependent; in general, mala rats are more susceptible than females, with the NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 48 &4/ 0. 0. 'Jo earliest lesions appearing at about 3 months of age (Goldstein et al., 1988). Rats were 4.5 months old at the end of the current studies. Renal tubular epithelial regeneration was present in many dosed and control animals of all strains; however, only female F344/N rats exposed to 1,2-dichloroethane in drinking water had a higher incidence of kidney lesions than controls. The degree of severity was not increased, however, and was minimal even in the highest dose group. Administration of up to 8,000 ppm 1,2-dichloroethane in drinking water resulted in greater toxicity to B6C3Fi mice than to rats. Nine of 10 female mice exposed to 8,000 ppm 1,2-dichloroethane died before the end of the study. The estimated daily intake of 1,2-dichloroethane in mice (with no corrections made for spillage) administered 8,000 ppm 1,2-dichloroethane was approximately 4,200 mg/kg in males and 4,900 mg/kg in females. These intake levels are approximately tenfold greater than the reported LD50 of 1,2dichloroethane administered by gavage (489 mg/kg for male mice and 413 mg/kg for female mice) (Munson at al.( 1982). The estimated daily intake of 1,2dichloroethane was considerably higher for mice than for rats receiving the same concentrations in drinking water. Mice typically consume more water than rats on a milligram per kilogram body weight basis, and palatability did not reduce water consumption by mice. Based on organ weight changes, the target organs for male and female B6C3Fi mice exposed to 1,2-dichloroethane in drinking water were the liver and 49 1,2- Dichloroethane, NTP TOX 4, Draft 6/89 SAL 000066911 NOT FOR DISTRIBUTION OR ATTRIBUTION 3oara Draft kidney. However, histopathologic changes were limited to protein casts, mineralisation, karyomegaly, and regeneration in the renal tubiiles of male mice. The regenerative lesions were similar to those observed in rats; however, such lesions are generally less common in mice than in rats. Although significant increases were observed in kidney weights of most exposed female mice, regeneration was detected in only one mouse. Long-term studies have shown that 1,2-dichloroethane administered by gavage causes neoplasms in the mammary gland, endometrium, and lungs (but not in the kidney) in B6C3?i mice (MCI, 1978); inhalation exposure of Swiss mice resulted in no carcinogenic effects (Haltoni et al., 1980). The differing results of the two long-term studies have been attributed to a difference in responsiveness in the test strains and to the different routes of administration (Hooper et al., 1980). The results from an short-term study on B8C3Fi mice indicated that 1,2dichloroethane is capable of inducing single-strand breaks and/or alkalilabile lesions in hepatic DMA when administered by intraperitoneal injection or by gavage, but not after inhalation exposure to comparable doses (Storer et al., 1984); this suggests that the liver is more likely to be a target organ when 1,2-dichloroethane is administered orally or parenterally than when administered by inhalation. The current drinking water studies in B6C3Fi mice demonstrated increases in liver weights in mice receiving drinking water containing 1,2-dichloroethane, although histologic lesions were not observed. In addition, lesions were observed in the kidney, which had not previously been identified as a target organ in mice. NOT FOR DISTRIBUTION OR ATTRIBUTION 1.2-Dichloroethane, NTPTOX 4, Draft 6/89 50 o00066t?^-2 (5Ai- Board Dn;'* 1.2- Dichloroethane administered at up to 8,000 ppm in drinking water for 13 weeks was relatively nontoxic for F344/N, Sprague Dawley, and Osborne-Mendel rats. Administration of the same drinking water concentrations of 1,2dichloroethane to B6C3Fi mice resulted in greater toxicity; 9/10 female mice exposed to 8,000 ppm 1,2-dichloroethane died before the end of the study. The estimated daily intake (milligram per kilogram per day) of 1,2dichloroethane in mice was about tenfold greater than in rats. Based on organ weight increases, the liver and kidney appeared to be target organs in both rats and mice, although histologic evidence of toxicity was found only in the kidney of female F344/N rats (minimal) and male B6C3Fi mice. Because of limitations in the solubility and palatab ility of 1,2dichloroethane, it was not possible to obtain a high enough dose in drinking water to see biologically significant toxic effects in rats. Based on mortality and chemical-related lesions, the no-effeet levels for 1,2dichloroethane administered by gavage to F344/N rats were 120 mg/kg for males and 150 mg/kg for females. For B6C3Fi mice, the no-effect levels for 1.2- dichloroethane in drinking water were 2,000 ppm for males, based on kidney lesions, and 4,000 ppm for females, based on mortality. V. REFERENCES Alumot, E.; Hachtomi, E.; Handel, E.; Holstein, P.; Bondi, A.; Heriberg, M. (1976) Tolerance and acceptable daily intake of chlorinated fumigants in the rat diet. Food Cosmet. Toxicol. 14:105-110. Anders, M.w.; Jakobson, I. (1985) Biotransformation of halogenated solvents. Scand. J. Work Environ. Health ll(Suppl. l):23-32. 51 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 SA/~ 000066913 NOT FOR DISTRIBUTION OR ATTRIBUTION Board Drift Barsoum, G.S.; Saad, K. (1934) Relative toxicity of certain chlorine derivations of the aliphatic series. Q. J. Pharm. Pharmacol. 7:205-214. Bignami, M.; Cardamone, G.; Comba, P.; Ortall, V.A.; Morpurgo,'G.; Carere, A. (1977) Relationship between chemical structure and mutagenic activity in some pesticides: The use of Salmonella, typhimurium and Aspergillus nidulans. Hutat. Res. 46:243-244. Boorman, G.A.; Montgomery, C.A., Jr.; Eustis, S.L.; Wolfe, H.J.; McConnell, E.E.; Hardisty, J.7. (1985) Quality assurance in pathology for rodent carcinogenicity studies. Milman, B.; Weisburger, E., Eds.: Handbook of Carcinogen Testing. Park Ridge, NJ: Noyes Publications, pp. 345-357. Brem, H.; Stein, A.B.; Rosenkranz, H.S. (1974) The mutagenicity and DNAmodifying effect of haloalkanes. Cancer Res. 34:2576-2579. . Cetnarovicz, J. (1959) Experimental and clinical investigations on the action of dichloroethane. Folia Med. Cracov. 1:169-192. Crespi, C.L.; Seixas, G.M.; Turner, T.R.; Ryan, C.G.; Penman, B.W. (1985) Mutagenicity of 1,2-dichloroethane and 1,2-dibromoethane in two human lymphoblastoid cell lines. Mutat. Res. 142:133-140. Davidson, I.W.F.; Sumner, D.O.; Parker, J.C. (1982) Ethylene dichloride: A review of its metabolism, mutagenic and carcinogenic potential. Drug Chem. Toxicol. 5:319-388. Drury, J.S.; Hammons, A.S. (1979) Investigations of Selected Environmental Pollutants: 1,2-Dichloroethane. Contract No. EPA 560/2-78-006. Oak Ridge, TN: Environmental Protection Agency Oak Ridge National Laboratory. Dunn, O.J. (1964) Multiple comparisons using rank sums. Technometrics 6:241-252. Elfers, L.A. (1979) Monitoring of Ambient Levels of EDC in the Vicinity of EDC Production and User Facilities. Contract No. EPA 600/4-79-029. Research Triangle Park, NC: U.S. Environmental Protection Agency. Ewing, B.B.; Chian, E.S.K.; Cook, J.C.; Evans, C.A.; Hopke, P.K.; Perkins, E.G. (1977) Monitoring to Detect Previously Unrecognized Pollutants in Surface Waters. Contract No. EPA 560/6-77-015. Washington, DC: U.S. Environmental Protection Agency, pp. 63-64, 73. Gold, L.S. (1980) Human exposures to ethylene dichloride. Ames, B,; infante, P.; Reitz, R., Eds.: Ethylene Dichloride: A Potential Health Risk? Banbury Report 5. Cold Spring Harbor, NT: Cold Spring Harbor Laboratory, pp. 209-225. Goldstein, R.S.; Tarloff, J.B.; Hook, J.B, (1988) Age-related nephropathy in laboratory rats. Fed. Am. Soc. Exp. Biol. J. 2:2241-2251. NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 52 sal 000669l4 Boara Drift Guengerich, F.P.; Crawford, W.M., Jr.; Domoradzki, J.Y.; MacDonald, T.L.; Watanabe, P.G. (1980) In vitro activation of 1,2-dichloroethane by microsomal and cytosolic enzymes. Toxicol. Appl. Pharmacol. 55:303-317. Heppel, L.A.; Heal, P.A.; Perrin, T.L.; Endicott, K.M.; Porterfield, V.T. (1946) The toxicology of 1,2-dichloroethane (ethylene dichloride). V. The effects of daily inhalations. J. Ind. Hyg. Toxicol. 28:113-120. Hofmann, H.T.; Birnstiel, H.; Jobst, P. (1971) Zur Inhalationstoxicitat von 1,1- und l,2-Dichlorathan. Arch, Toxikol. 27:248-265. Hooper, K.; Gold, I.; Ames, B. (1980) The carcinogenic potency of ethylene dichloride in two animal bioassays: A comparison of inhalation and gavage studies. Ames, B.; Infante, P.; Reitz, R., Eds.: Ethylene Dichloride: A Potential Health Risk? Banbury Report 5. Cold Spring Harbor, MY: Cold Spring Harbor Laboratory, pp. 65-81. Hueper, W.C.; Smith, C. (1935) Fatal ethylene dichloride poisoning. Am. J. Med. Sci. 189:778-784. Inskeep, P.B.; Koga, H.; Cmarik, J.L.; Guengerich, F.P. 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(1980) The micronucleus test as a part of a short term mutagenicity test program for the prediction of carcinogenicity evaluated by 143 agents tested. Mutat. Res. 75:191-202. Jonckheere, A. (1954) A distribution-free k-sample test against ordered alternatives. Biometrika 41:133-145. Jakobson, I.; Wahlberg, J.I.; Holmberg, B.; Johansson, G. (1982) Uptake via the blood and elimination of 10 organic solvents following epicutaneous exposure of anesthetized guinea pigs. Toxicol. Appl. Pharmacol. 63:181-187. 53 1,2-Dichloroethane, NTPTOX4, Draft 6/89 ,0*6<n5 5^- ooo NOT FOR DISTRIBUTION OR ATTRIBUTION Board Drift Kellam, R.G.; Dusetzina, M.G. (1980) Human exposure to ethylene dichloride: Potential for regulation via EPA's proposed airborne carcinogen policy. Ames, a.; Infante, P.; Reitz, R., Eds.: Ethylene Dichloride: A Potential Health Risk? Banbury Report 5. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory, pp. 265-274. King, M.-T.; Beikirch, H.; Eckhardt, K.; Gocke, E.; Wild, . (1979) Mutagenicity studies with X-ray contrast media, analgesics, antipyretics, antirheumatics and some other pharmaceutical drugs in bacterial, Drosophila and mammalian test systems. Mutat. Res. 66:33-43. Klaunig, J.E.; Ruch, R.J.; Pereira, M.A. (1986) Carcinogenicity of chlorinated methane and ethane compounds administered in drinking water to mice. Environ. Health Perspect. 69:89-95. Kozik, I.V. (1957) Some problems of occupational hygiene in the use of dichloroethane in the aircraft industry. Gig. Tr. Prof. Zabol. 1:31-38. Kramers, P.G.; Bissumbhar, B. (1983) Role of exposure period in applying gaseous mutagens to Drosophila, as exemplified by 1,2-dichloroethane and methylbromide. Mutat. Res. 113:272. Lane, R.W.; Riddle, B.L.; Borzelleca, J.F. (1982) Effects of 1,2dichloroethane and 1,1,1-trichloroethane in drinking water on reproduction and development in mice. Toxicol. Appl. Pharmacol. 63:409-421. Larionov, V.G.; Kokarovtseva, M.G. (1976) Morphological constitution of peripheral blood in intoxication with dichloroethane and its metabolites. Actual Problems of Pesticide Application in Different Climatographic Zones. Yerevan: Aiastan Publishers, pp. 131-133. Letkiewicz, ?.; Johnson, P.; Colman, J.; et al. (1982) Occurrence of 1,2Dichloroethane in Drinking Water, Food and Air. EPA Contract No. 68-016185, Task 11. Lochhead, H.B.; Close, H.P. (1951) Ethylene dichloride plastic cement: A case of fatal poisoning. J. Am. Med. Assoc. 146:1323. Maltoni, C.; Valgimigli, L.; Scamato, C. (1980) Long-term carcinogenic bioassays on ethylene dichloride administered by inhalation to rats and mice. Ames, B.; Infante, P.; Reitz, R., Eds.: Ethylene Dichloride: A Potential Health Risk? Banbury Report 5. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory, pp. 3-29. Maronpot, R.R.; Boorman, G.A. (1982) Interpretation of rodent hepatocellular proliferative alterations and hepatocellular tumors in chemical safety assessment. Toxicol. Pathol. 10:71-80. McCann, J.; Simmon, V.; Streitwieser, D.; Ames, B.N. (1975) Mutagen icity of chloroacetaldehyde, a possible metabolic product of 1,2-dichloroethane (ethylene dichloride), chloroethanol (ethylene chlorohydrin), vinyl chloride and cyclophosphamide. Proc. Natl. Acad. Sci. (USA) 72:3190-3193. o 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 54 "G NOT FOR DISTRIBUTION OR ATTRIBUTION OOo, Board Draft McCollister, D.D.; Hollingsworth, R.L.; Oyen, F.; Rowe, V.K. (1956) Comparative inhalation toxicity of fumigant mixtures. Arch. Ind. Health 13:1-7. Menschick, H. (1957) Acute inhalation intoxications by symmetric dichloroethane. Arch. Gewerbepathol. Gewerbehyg. 15:241-252. Hitoma, C.; Steeger, T.; Jackson, S.E.; Wheeler, K.P.; Rogers, J.H.; Milman, H.A. (1985) Metabolic disposition study of chlorinated hydrocarbons in rats and mice. Drug Chem. Toxicol. 8:183-194. 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(1980) Teratogenicity and reproduction studies in animals inhaling ethylene dichloride. Ames, B.; Infante, P,; Reitx, R., Eds.: Ethylene Dichloride: A Potential Health Risk? Banbury Report 5. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory, pp. 149-161. Reitz, R.H.; Fox, T.R.; Domoradzki, J.Y.; Quast, J.F.; Langvardt, P.; Watanabe, P.G. (1980) Pharmacokinetics and macromolecular interactions of ethylene dichloride: Comparison of oral and inhalation exposures. Ames, B.; Infante, P.; Reitz, R., Eds.: Ethylene Dichloride: A Potential Health Risk? Banbury Report 5. Cold Spring Harbor, MY: Cold Spring Harbor Laboratory, pp. 135-144. Reitz, R.H.; Fox, T.R.; Ramsey, J.C.; Quast, J.F.; Langvardt, P.W.; Watanabe, P.G. (1982) Pharmacokinetics and macromolecular interactions of ethylene dichloride in rats after inhalation or gavage. Toxicol. Appl. Pharmacol. 62:190-204. Rosenkranz, H.s. (1977) Mutagenicity of halogenated alkanes and their derivatives. Environ. Health Perspect. 21:79-84. 55 1,2-Dichloroethane, NTP TOX 4, Draft 6/89 ,00' NOT FOR DISTRIBUTION OR ATTRIBUTION Board Draft Seufert, F.3.; Brown, P. Oatway, J.A.; Bornstein, M. ; Ostrowski, W. ; Horne, R. (1980) L.J-Oichloroethane Technical Control Options Analysis. EPA Contract Ho. 68-01-5960. Bedford, MA: CGA Corporation. Shakarnis, V.F. (1969) induction of X chromosome nondisjunction and recessive sex-linked lethal mutations in females of Drosophila melanogaster by 1,2-dichloroethane. Genetika 5:89-95. Shirley, E. (1977) A non-parametric equivalent of Williams' test for contrasting increasing dose levels of a treatment. Biometrics 33:386-389. Shmuter, L.M. (1977) Effect of chronic exposure to low concentrations of chlorinated hydrocarbons of the ethane series on specific and non-specific reactivity of animals in vivo. Gig. Tr. Prof. Zabol. 8:38-42. Simmon, V.F.; Kauhanen, K.; Tardiff, R.G. (1977) Mutagenic activity of chemicals identified in drinking water. Scott, D.; Bridges, B.A.; Sobels, F.H., Eds.: Progress in Genetic Toxicology. Amsterdam: Elsevier, pp. 249258. Singh, H.B.; Salas, L.J.; Stiles, R.E. (1983) Selected man-made halogenated chemicals in the air and oceanic environment. J. Geophys. Res. 88:36753683. Sopikov, H.G.; Gorshunova, A.I. (1979) Investigation of the intake, distribution and excretion of ethylene dichloride in rats. Gig. Tr. Prof. Zabol. 4:36-40. Spencer, H.C.; Rowe, V.K.; Adams, E.M.; McCollister, D.D.; Irish, D.D. (1951) Vapor toxicity of ethylene dichloride determined by experiments on laboratory animals. Arch. Ind. Byg. Occup. Med. 4:482-493. Spreafico, 7.; Zuccato, E.; Marcucei, 7.; Sironi, M.; Paglialunga, S.; Madonna, M.; Mussini, E. (1980) Pharmacokinetics of ethylene dichloride in rats treated by different routes and its long-term inhalatory toxicity. Ames, B.; Infante, P.; Reits, R., Eds.: Ethylene Dichloride: A Potential Health Risk? Banbury Report 5. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory, pp. 107-129. Storer, R.D.; Jackson, H.M.; Conolly, R.B. (1984) In vivo genotoxicity and acuta hepatotoxieity of 1,2-dichloroethane in mice: Comparison of oral, intraperitoneal, and inhalation routes of exposure. Cancer Res. 44:42674271. Sundheimer, D.W.; White, R.D.; Brendel, K.; Sipes, I.G. (1982) The bioactivation of 1,2-dibromoethane in rat hepatocytes: Covalent binding to nucleic acids. Carcinogenesis 3:1129-1133. Suveev, I.M.; Babichenko, M.E. (1969) On the clinic and cure of acute intoxication with dichloroethane vapours. Gig. Tr. Prof. Zabol. 13:50-51. CD NOT FOR DISTRIBUTION OR ATTRIBUTION 000669 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 56 <z CD Board Dra:: Symons, J.M.; Bellar, T.A.; Carswell, J.K.; DeMarco, J.; Kropp, K.L.; Robeck, G.G.; Seeger, D.R.; Slocum, C.J.; Smith, B.L.; Stevens, A.A. ( 1975) National Organics Reconnaissance Survey for halogenated organics. J. Am. Water Works Assoc. 67:634-647. Theiss, J.C.; Stoner, G.D.; Shimkin, M.B.; Weisburger, E.K. (1977) Test for carcinogenicity of organic contaminants of United States drinking waters by pulmonary tumor response in strain A mice. Cancer Res. 37:2717-2720. Torkelson, T.R.; Rowe, V.K. (1981) Halogenated aliphatic hydrocarbons containing chlorine, bromine, and iodine. Clayton, G.D.; Clayton, F.E., Eds: Patty's Industrial Hygiene and Toxicology, 3rd rev. ed., Vol. 2B. York: John Wiley i Sons, Inc., p. 3491. New Troisi, F.M.; Cavallazzi, D. (1961) A fatal case of poisoning from inhalation of dichloroethane vapours. Med. Lav. 52:612-618. Tsuruta, H. (1975) Percutaneous absortion of organic solvents. I. Comparative study of the in vivo percutaneous absorption of chlorinated solvents in mice. Ind. Health 13:227-236. Urusova, T.P. (1953) The possible presence of dichloroethane in human milk with exposure in industrial conditions. Gig. Sanit. 18:36-37. U.S. Environmental Protection Agency (USEPA) (1985) Health Assessment Document for l,2-Dichloroethane (Ethylene Dichloride). Contract No. EPA/600/8-84/006F. Washington, DC: USEPA, Office of Health and Environmental Assessment. U.S. International Trade Commission (USITC) (1987) Synthetic Organic Chemicals 1986. Publication No. 2009. Washington, DC: U.S. Government Printing Office, p. 212. Van Duuren, 8.L.; Goldschmidt, B.M.; Loewengart, G.; Smith, A.C.; Melchionne, S.; Seldman, I.; Roth, D. (1979) Carcinogenicity of halogenated olefinic and aliphatic hydrocarbons in mice. J. Natl. Cancer Inst. 63:14331439. Van Esch, G.J.; Kroes, R.; Van Logten, M.J.; Den Tonkelaar, E.M. (1977) Ninety-Day Toxicity Study with 1,2-Dichloroethane (DCE) in Rats. Report No. 195/77 Al. Tox. Bilthoven, Netherlands: National Institute of Public Health and Environmental Hygiene. Versar, Inc. (1975) Identification of Organic Compounds in Effluents Industrial Sources. Final Report to EPA Office of Toxic Substances. Contract No. EPA 560/3-75-002. Washington, DC: U.S. Environmental Protection Agency. from Vosovaya, M.A. (1974) Development of posterity of two generations obtained from females subjected to the action of dichloroethane. Gig. Sanit. 7:2528. NOT FOR DISTRIBUTION OR ATTRIBUTION 57 1.2-Dichloroethane, NTPTOX 4, Draft 6/89 6691? SAL oo i Board Ora:' Vosovaya, M.A. (1977) Effect of dichloroethane on the reproductive cycle and embryogenesis in experimental animals. Akush. Ginekol. 2:57-59. Weiss, p. (1957) Lethal oral intoxications by dichloroethane. Geverbehygiene 15:253-264. Withey, J.R.; Kaxpinski, K. (1985) The fetal distribution of some aliphatic chlorinated hydrocarbons in the rat after vapor phase exposure. Biol. Res. Pregnancy 6:79-88. Yllner, S. (1971) Metabolism of 1,2-dichloroethane-1<C in the mouse. Acta Pharmacol. Toxicol. 30:257-265. NOT FOR DISTRIBUTION OR ATTRIBUTION 00066 1,2-Dichloroethane, NTP TOX4, Draft 6/89 58 O c-j >. < CO APPENDIX A ORGAN WEIGHT, HEMATOLOGIC, AND SERUM CHEMICAL DATA IN THE THIRTEEN-WEEK STUDIES OF 1,2-DICHLOROETHANE TABLE A1 TABLE A2 TABLE A3 TABLE A4 TABLE A5 TABLE A6 TABLE A7 TABLE A8 TABLE A9 TABLE A10 TABLE All TABLE A12 TABLE A13 TABLE A14 PACE ABSOLUTE ORGAN WEIGHTS FOR F344/N RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1,2-DICHLOROETHANE A-2 ORGAN WEIGHT TO BODY WEIGHT RATIOS FOR F344/N RATS IN THE THIRTEENWEEK DRINKING WATER STUDIES OF 1,2-DICHLOROETHANE A-2 HEMATOLOGIC AND SERUM CHEMICAL DATA FOR MALE F344/N RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF U-DICHLOROETHANE A-3 ABSOLUTE ORGAN WEIGHTS FOR SPRAGUE DAWLEY RATS IN THE THIRTEENWEEK DRINKING WATER STUDIES OF U-DICHLOROETHANE A-5 ORGAN WEIGHT TO BODY WEIGHT RATIOS FOR SPRAGUE DAWLEY RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF U-DICHLOROETHANE A-5 HEMATOLOGIC AND SERUM CHEMICAL DATA FOR MALE SPRAGUE DAWLEY RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1,2-DICHLOROETHANE A-6 ABSOLUTE ORGAN WEIGHTS FOR OSBORNE-MENDEL RATS IN THE THIRTEENWEEK DRINKING WATER STUDIES OF U-DICHLOROETHANE A-8 ORGAN WEIGHT TO BODY WEIGHT RATIOS FOR OSBORNE-MENDEL RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF U-DICHLOROETHANE A-9 HEMATOLOGIC AND SERUM CHEMICAL DATA FOR MALE OSBORNE-MENDEL RATS IN THE THIRTEEN.WEEK DRINKING WATER STUDIES OF U-DICHLOROETHANE A-10 ABSOLUTE ORGAN WEIGHTS FOR F344/N RATS IN THE THIRTEEN-WEEK GAVAGE STUDIES OF U-DICHLOROETHANE A-12 ORGAN WEIGHT TO BODY WEIGHT RATIOS FOR F344/N RATS IN THE THIRTEENWEEK GAVAGE STUDIES OF 1,2-DICHLOROETHANE A-12 HEMATOLOGIC AND SERUM CHEMICAL DATA FOR MALE F344/N RATS IN THE THIRTEEN-WEEK GAVAGE STUDIES OF 1,2-DICHLOROETHANE A-13 ABSOLUTE ORGAN WEIGHTS FOR B6C3Ft MICE IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1,2-DICHLOROETHANE A-15 ORGAN WEIGHT TO BODY WEIGHT RATIOS FOR B6C3Ft MICE IN THE THIRTEENWEEK DRINKING WATER STUDIES OF 1,2-DICHLOROETHANE A-16 NOT FOR DISTRIBUTION OR ATTRIBUTION A-l 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 S/9/. 0()06,$oo. TABLE Al. ABSOLUTE ORGAN WEIGHTS FOR F344/N RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1JJ-DICHLOROETHANE (a) Control 300 ppm 1,000 ppm 2,000 ppm 4,000 ppm 8,000 ppm MALE Brain 1.959 Heart 1,044 Right kidney 1.232 Liver 15,450 Lung 1.731 Right testis 1.462 Thymus 285 23 24 4- 48 660 41 15 14 1.927 1,077 1,345 16.500 1.864 1.460 304 30 23 38 540 74 33 17 FEMALE Brain 1,795 16 Heart 633 17 Right kidney 739 26 Liver 6.829 154 Lung 1,203 35 Thymus 242 9 1,817 20 654 12 814 16 7,268 179 1.488 169 247 7 1.958 1,062 "1,433 16.960 (b) 1,824 1,467 287 24 1.954 13 1.078 28 *1,523 570 *17.840 97 1.770 19 1.462 3 302 21 1.930 24 *991 15 **1,451 250 16,050 61 1.634 24 1,476 15 307 30 1,908 9 **927 18 "1,377 330 14.760 72 1,632 19 1,422 21 258 t 28 12 t 22 340 47 30 13 1.786 665 "885 **7,627 1,353 242 28 9 16 177 126 13 1,772 17 667 13 **845 17 7,278 165 1,175 61 221 16 1.801 648 "932 7,551 1.243 236 , 16 3 15 171 35 13 1,773 37 643 12 "923 15 7,134 147 1.224 50 234 12 (a) Mean standard error in milligrams for groupa of 10 animals unless otherwise specified; P values vs. the controls by Dunn's test < Dunn, 1964) or Shirley's test (Shirley, 1977). (b) Lungs of nine animals were weighed. *P<0.05 **P<0.01 TABLE A3. ORGAN WEIGHT TO BODY WEIGHT RATIOS TOR F344/N RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF U-DICHLOROETHANE (a) Control 500 ppm 1,000 ppm 2.000 ppm 4,000 ppm 8,000 ppm MALE Body weight (grams) Brain Heart Right kidney Liver Lung Right testis Thymus FEMALE Body weight (grams) Brain Heart Right kidney Liver Lung Thymus 363 12.0 5.4 0.15 2.9 0.08 3.4 0.16 42.9 2.17 4.8 0.18 4.1 0.11 0.8 0.05 354 6.9 355 4.5 355 2.8 *327 2.8 **300 4.3 5.5 0.11 3.0 0.06 3.8 0.08 46.5 0.95 5.3 0.23 4.1 t 0.05 0.9 0.04 5.5 0.07 3.0 0.04 **4.0 0.09 47.7 1.37 (b>5.2 0.29 4.1 0.07 0.8 0.02 5,5 0.08 3.0 0.07 "4.3 0.04 **50.2 0.49 5.0 t 0.17 4.1 0.05 0.9 0.05 "5.9 0.06 3.0 0.03 *4.4 0.06 *49.1 0.79 5.0 0.21 "4.5 0.05 0.9 0.07 *6.4 0.11 3.1 0.03 "4,6 0.07 *49.2 0.85 5.5 0.18 **4.7 0.08 0.9 0.04 194 2.4 9.3 0.15 3.3 0.09 3.8 t 0.13 35.3 0.85 6.2 0.15 1.3 0.04 199 2.9 9.2 0.13 3.3 0.05 4.1 0.07 36.6 0.60 7.5 0.82 1.2 0.04 213 t 10.1 8.5 0.30 3.2 0.12 4.2 0.17 36.3 1.57 6.4 0.60 1.2 0.09 196 2.4 193 1.3 185 2.3 9.0 0.07 3.4 0.05 "4.3 0.07 37,2 0.75 6.0 0.27 1.1 0.08 9.4 0.09 3.4 0.05 "4.8 0.09 "39.2 0.94 6.5 0.17 1.2 0.07 9.6 0.14 *3.5 0.05 *5.0 0.04 *38.5 0.61 6.6 0.23 1.3 0.06 (a) Mean standard error in milligrams per gram for groups of 10 animals unless otherwise specified; P values vs. the controls by Dunn's test (Dunn, 1964) or Shirley's test (Shirley, 1977) < b) Lungs of nine animals were weighed. *P<0.05 **P<0.01 I ( ( j i i Ii ( ( ( i 1 ( NOT FOR DISTRIBUTION OR ATTRIBUTION ^00(569 1,2-Dichloroethane, NTPTOX4, Draft 6/89 A-2 o -j <r TABLE A3. HEMATOLOGIC AND SEBUM CHEMICAL DATA FOR MALE F344/N RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1J-DICHLOROETHANE (a) Analysis Day Control 2,000 Pptn 4,000 ppm 8,000 ppm Number etaminedlb) Leukocytes (1,000/pJ) 3 7 14 45 90 Hematocrit (percent) 3 7 14 45 90 Hemoglobin (g/dl) 3 7 14 45 90 Mean corpuscular hemoglobin (pg) 3 7 14 45 90 Mean cell hemoglobin concentration I g/dl) 3 7 14 45 90 Mean cell volume (p3) 3 7 14 45 90 Platelets 11,000/pl) 3 7 14 45 90 Erythrocytes (10/yil> 3 7 14 45 90 Alkaline phosphatase (lU/liter) 3 7 14 45 90 Alanine aminotransferase UU/liter) 3 7 14 45 90 3 20.7 + 4.28 (e) 7.4 0.96 6 9 0.61 7 7 0.48 7.2 0.35 40.7 1.18 (e) 41.2 0.71 43.5 0.50 45 4 1.03 46.5 0.37 13.7 1 13 (e) 14.8 0.17 15.0 + 0.12 17.0 + 0.15 16.8 0.07 21.5 1.24 le) 22.5 0.29 22.2 0.18 20.0 0.41 18.9 0.16 33.4 2.28 (e) 36.0 0.43 34.4 0.24 37.6 0.85 36.2 t 0.33 65.4 + 2.26 (e)62.6 0.87 64.5 0.53 53.1 0.64 52.0 0.57 (e)979 + 58.3 (e) 861 34.8 836 33.6 (e)540 25.7 471 24.9 6.3 0.36 (e)6.6 0.11 6.8 0.08 8.6 0.17 8.9 0.10 638 20.2 690 44.2 631 21.5 330 12.5 290 5.7 (f) 50.0 4.40 (e) 38.7 + 1.74 (e) 37.6 2.00 (e) 48.3 3.24 (e) 73.1 6.07 10 fc) 15.1 5.64 8.9 1.18 6.6 0.34 8.1 0.21 8.8 0.82 ic) 40.1 0.56 41.8 0.52 43.8 0.38 45.5 1.04 46.3 + 0.43 (c) 14.3 0.20 14.9 0.12 15.2 0.10 17.1 0.13 16.7 0.08 (0 22.6 + 0.68 22.0 0.45 21.5 0.35 19.3 0.58 18.4 0.24 (0 35.7 0.49 35.7 0.27 34.7 0.28 37.9 + 0.93 36.0 + 0.37 (0 63.3 1.89 62.0 1.00 *62.1 0.85 52.5 0.58 50.9 0.23 (0 943 53.0 831 27.4 775 25.6 550 17.8 488 24.2 (06.4 0.23 6.8 0.14 7.1 0.12 8.7 0.22 9.1 0.10 <e>614 *(0 590 561 316 263 41.7 10.7 19.8 .11.9 8.0 (g)41.6 + 1.08 (e)37.6 1.29 (g> 35.8 2.31 47.8 3.40 (c) 61.0 + 3.87 10 10 id) 24.9 6.89 (o 7.9 0.54 6.4 0.30 7 6 0.19 7.6 0.24 id) 40.5 0.61 *(c)44.1 0.87 44.3 0.33 47.1 0.29 46.5 044 Id) 14.0 0.23 (c) 15.5 0.28 15.3 0.14 17.4 0.07 16.7 0.12 6.9 0.47 7.2 0.39 6.9 0.27 7.0 0.36 6.9 0.27 "44.7 + 0.66 41.5 0.38 43.5 0.51 46.8 0.70 46.9 0.51 *15.4 0.17 15.1 + 0.19 15.2 0.19 16.8 0.07 16.8 0.07 (d) 22.4 0.54 (c) 22.0 0.25 21.5 0.26 19.6 0.19 *18.2 0.10 (d) 34.6 0.43 lc)35.1 0,25 34.5 + 0.29 36.8 0.25 36.0 0.27 *20,7 0.25 21.2 0.12 "20.7 0.14 *18.8 0.27 *18.1 0.12 34.5 0.32 36.5 0.27 35.0 > 0.24 36.0 0.46 35.8 0.31 (d) 65.0 2.06 (0 62.7 0.97 62.4 0.92 53.3 0.30 50.6 0.34 60.0 0.33 "57.9 + 0.35 *59.4 0.31 52.3 0.33 50.3 0.37 (e) 1,019 57.2 (0 821 27.5 738 37.1 543 7.6 488 17.2 (c) 872 18.7 738 20.8 706 24.3 544 6.6 508 19.2 (d)6.3 + 0.24 (c)7.0 0.16 7.1 0.11 8.9 0.07 9.2 0.08 7.4 0.13 *7,2 0.09 7.3 0.07 *8.9 0,14 "9.3 0.07 !d)614 24.3 (0 553 21.5 594 30.5 329 7.4 278 9.0 609 19.5 *(0 562 32.1 (c) 557 19.1 338 17.1 285 9.6 (h) 36.8 2.43 (f) 40.0 3.29 (d) 36.9 + 2,86 (e) 32.0 1,36 <d) 35.5 1.35 (d) 36.8 1.62 (043.1 2.40 48.7 + 3.32 *<e)54.0 3.73 "(e) 54.3 2.20 NOT FOR DISTRIBUTION OR ATTRIBUTION A-3 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 SAL 000066923 TABLE A3. HEMATOLOGIC AND SERUM CHEMICAL DATA FOR MALE F344/N RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF 1JJ-DICHLOROETHANE (Continued) Analysis Day Blood urea nitrogen img/dl) 3 7 14 45 90 Creatine kinase i lU/liter) 3 7 14 45 90 Sorbitol dehydrogenase i IU/liter) 3 7 14 45 90 Control 2.000 ppm 4.000 ppm 8,000 ppm <i) 14.0 0.00 ih) 17.3 + 2.17 (e) 19.4 * 1.46 (0 24.7 1.76 >) 16.4 + 0.90 "(d) 23.4 1.51 id) 25.1 2.16 27.9 1.54 id) 20.8 0.88 20.5 + 0.67 <c) 986 225 587 125 381 + 53 562 49 341 t 33 (0 605 + 96 (0 598 42 400 28 478 49 351 t 39 (e) 6.6 + 0.65 (e) 8.6 0.48 Ig) 9.2 0.97 10.3 + 0.82 le) 22.0 9.75 (e)7.4 1.19 109.9 0.77 (c) 8.7 0.24 <d) 11.9 1.61 12.0 1.14 *(f) 26.8 2.00 (g) 19.2 1.85 (0 25.1 1.40 1"(e) 30.9 4.23 *(0 24.7 1.44 *(f) 21.8 1 97 (el 25.4 2.08 25.7 4- 1 37 21.3 0.60 21,3 + 0.90 695 + 86 803 t 118 351 34 424 33 315 21 718 129 Ic) 504 39 374 48 441 > 48 320 12 if) 11.7 '4.94 le) 9.1 0.60 (d) 9.4 0.32 (e) 11.3 0.84 10.0 0.56 (e) 8.7 0.52 (d) 12.3 2.84 *10.2 0.33 13.5 1.15 10.4 1.16 (a) Mean standard error; P values vs, the controls by Dunn's test! Dunn, 1964) or Shirley's test (Shirley, 1977). (b) Unless otherwise specified Ic) Nine animals were examined. (d) Eight animals were examined. (e) Seven animals were examined. ID Six animals were examined. Ig) Five animals were examined. (h) Four animals were examined. (i) Three animals were examined. P<0.05 *P<0.01 NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2-Dichloroethane, NTPTOX4, Draft 6/89 A-4 T n: Os "0 -a o o o <E CO TABLE A4. ABSOLUTE ORGAN WEIGHTS FOR SPRAGUE DAWLEY RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF U-DICHLOROETHANE (a) Control 300 ppm 1,000 ppm 2,000 ppm 4,000 ppm 8.000 ppm male Brain 2.089 37 2.139 41 2,125 23 2.117 22 2.105 25 Heart 1.847 85 1.729 41 *1,623 t 54 *1.597 54 * *1,579 55 Right kidney 1,871 74 1,943 59 1,954 58 1.856 74 2.000 52 Liver Lung Right testis 18.480 790 20,080 590 18.810 570 20,100 790 19.970 490 2.468 83 2.728 161 2,407 127 2,558 95 2.342 68 1.821 48 1,728 + 53 1.843 54 1,756 53 1,704 35 Thymus 493 32 477 25 448 32 474 39 468 33 2,103 t 19 **1.566 55 2.008 55 19,230 560 2,220 96 1.825 34 485 34 FEMALE Brain 1,975 29 1.975 36 2.005 31 1,963 19 1,913 29 1,956 34 Heart 1,069 26 1.072 30 1,084 24 1.061 32 1.041 '27 1,085 36 Right kidney 1.030 36 *1,160 27 **1,221 28 **1.211 33 **1,208 50 **1.342 16 Liver 11,140 350 11.890 530 12.200 680 10,990 t 310 11,500 370 (b) 11,950 450 Lung 1.929 89 1.988 114 1,861 65 1.993 109 1.915 99 1,941 1135 Thymus 364 23 395 36 337 17 365 23 359 30 326 23 (a) Mean standard error in milligrams for group* of 10 animals unless otherwise specified; P values vs. the controls by Dunn's test (Dunn. 1964) or Shirley's test (Shirley, 1977). (b) Nine livers were weighed. *P<0.QS "PcO.Ol TABLE AS. ORGAN WEIGHT TO BODY WEIGHT RATIOS FOR SPRAGUE DAWLEY RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES Or 14-DICHLOROETHANE (a) Control 500 ppm 1,000 ppm 2,000 ppm 4,000 ppm 8.000 ppm male Body weight (grams) Brain Heart Right kidney Liver Lung Right testis Thymus FEMALE Body weight (grams) Brain Heart Right kidney Liver Lung Thymus 449 t 11.0 446 7.9 4.7 0.16 4.1 0.20 4.2 0.14 41.1 1.03 5.5 0.18 4.1 0.14 1.1 0.07 4.8 0.10 3.9 0.08 4.4 0.11 *45.0 1.15 6.1 0.33 3.9 0.11 1.1 0.06 431 7.0 432 11.3 436 7.9 4.9 0.09 3.8 0.11 *4.5 0.08 *43.6 t 0.75 5.6 0.29 4.3 0.12 1.1 0.08 4.9 0.17 3.7 0.09 4.3 0.11 **46.5 l.ll 5.9 0.19 4.1 0.17 1.1 0.07 4.9 t 0.12 *3.6 0.13 *4.6 0.11 "45.9 0.82 5.4 t 0.18 3.9 t 0.10 1.1 0.07 *414 9.2 *5.1 0.10 3.8 0.11 "4.9 0.11 **46.5 1.20 5.4 0.20 4.4 0.15 1.2 0.08 271 5.5 7.3 0.13 4.0 0.08 3.8 0.11 412 1.07 7.1 0.29 1.4 0.09 283 7.8 7.0 0.16 3.8 0.11 *4.1 0.09 42.0 1.49 7.1 t 0.42 1.4 0.13 287 6.4 7.0 0.16 3.8 0.11 *4.3 0.13 42,7 2.60 6.5 0.24 1.2 0.07 271 4.5 7.3 0.10 3.9 0.12 4.5 0.11 40.6 1.32 7.3 0.34 1.4 0.09 265 6.6 256 t 4.8 7,2 0.17 7.7 0.19 3.9 0.09 4.2 0.09 *4.6 0.16 **5.2 t 0.10 43.5 1.37 *(b)46.6 1.41 7.2 0.36 7.6 0.53 1.4 0.10 1.3 0.08 (a) Mean standard error in milligrams per gram for group* of 10 animals unless otherwise specified; P values vs. the controls by Dunn's test (Dunn, 1964) or Shirley's test (Shirley, 1977). (b) Nine livers were weighed. *P<0.05 **P<0.01 NOT FOR DISTRIBUTION OR ATTRIBUTION A-5 l,2-Dichlorothane, NTPTOX4, Draft 6/89 SAL 000066925 TABLE A8. HEMATOLOGIC A.VD SERUM CHEMICAL DATA FOR MALE SPRAGL'E DAWLEY RATS IN THE THIRTEEN.WEEK DRINKING WATER STUDIES OF 1,2-DlCHLORO ETHANE la) Analysis Day Control 2,000 ppm -4.000 ppm ' 8,000 ppm Leukocytes 11.000/pl) 3 7 14 45 90 Hematocrit 1 percent) ' 3 7 14 45 90 Hemoglobin I g/dl) 3 7 14 45 90 Mean corpuscular hemoglobin (pg) 3 7 14 45 90 Mean corpuscular hemoglobin 3 concentration (g/dl) 7 14 45 90 Mean cell volume (p3) 3 7 14 45 90 Platelets (1,000/pl) 3 7 14 45 90 Erythrocytes (10/pl) 3 7 14 45 90 Alkaline phosphates* ilU/liter) 3 7 14 45 90 Alanine aminotransferase (IU/liter) 3 7 14 45 90 10 6 0.71 (c) 15.0 1.31 11.7 0.86 9.8 > 0.59 9.1 0.24 38.1 + 0.76 (041.5 + 0.39 47.0 t 0.55 47 '. 0.68 48.0 0.73 13.6 0.18 (c) 14.2 0.16 15.3 0.11 17.0 0.13 17.0 0.24 24.5 0.57 (c)23.3 0.28 22.9 0.21 20.8 0.22 19.4 0.19 35.7 0.63 (0 34.1 0.21 32.6 0.40 36.0 0.38 35.5 0.32 68.5 t 0.65 (0 68.4 1.09 70.6 1.28 57.7 0.88 54.7 0.62 976 47.0 (b) 1,183 40.1 (d)990 46.6 755 27.1 758 16.7 5.6 0.13 (06.1 0.11 6.7 0.07 8.2 0.12 8.8 0.18 477 13.3 420 17.9 406 29.9 245 13.9 291 15.3 (c) 47.4 4.09 42.2 3.21 (d)39.7 2.22 50.4 3.50 50.7 3.11 10.8 0.74 *(d) 12.2 0.68 (b) 11.3 0.80 9.7 0.28 9.2 0.39 18.5 4.16 (c) 12.6 0.92 lb) 10.8 0.46 10.2 0.61 9.3 0.68 ib) 10.1 0.27 ib) 12.9 1.60 (e) 12.1 1.00 *8.3 4- 0.35 8.6 0.51 *'42.3 0.74 *40.7 0.80 **(b) -44.2 0.70 id) 43.2 0.93 (c)43.6 0.34 lb) 43.3 0.62 (b) 47.7 1.20 (b) 46.4 + 0.70 le) 47.0 0.80 47 3 0.88 46.9 0.37 47,5 0.42 49.2 0.29 48.8 0.51 47.9 0.49 "14.3 0.15 13.8 0,17 **lb) 15.1 0.21 (d) 14.6 t 0.23 (c) 14.8 0.09 (b) 14.6 0.21 (b) 15.6 t 0.16 (b) 15.1 4* 0.10 le)15.5 0.24 17.0 0.26 16.9 0.14 16.8 0.13 17.3 t 0.12 17.0 0.12 16.8 0.15 22.7 0.32 (d) 23.0 0.28 (b) 22.0 0.39 20.8 0.23 18.8 0.23 23.2 0.35 (c)23.0 0.33 (b) 22.8 0.18 20.8 0.12 18.9 0.17 *(b) 22.8 0.26 (b)22.4 0.42 (e) 22.7 0.64 20.2 0.17 18,7 0.24 33.9 0.43 (d>33.8 0.27 (b) 32.7 0.76 35.9 0.23 35.1 0.26 34.0 0.31 (c)34.0 0.25 lb) 32.7 0.39 36.1 0.28 35.0 0.27 (b)34.0 0.12 (b> 33.7 0.20 (e)33.0 0.66 35.3 0.22 35.1 0.24 67.0 0.70 (d)68.2 0.97 (b> 67.4 1.51 58.1 0.59 53.6 0.76 68.2 0.93 (O 68.0 1.13 (b) 70.0 1.22 57,8 0.25 54.2 0.47 (b) 66.9 0.85 (b)66.8 1.29 ie)69.3 2.59 57.1 0.57 53.3 0.83 1,060 89.0 949 49.7 838 36.4 775 28.4 699 27.4 1,080 37.6 *(0 957 44.3 894 31.7 (d) 751 17.6 723 21.8 (b) 1,031 47.9 **(d) 946 65.3 (b)904 58.4 (d)758 21.3 742 23.7 *6.3 0.12 id) 6.3 0.10 (b) 7.1 0.12 8.2 0.14 9.2 t 0.13 6.0 0.16 *(0 6.4 0.08 lb) 6.6 0.07 8.1 0.06 9.0 0.11 **(b> 6.6 0.15 (b) 6.5 0.19 (e) 6.8 0.25 8.3 0.07 9.0 0.14 430 14.5 428 t 24.5 416 12.1 228 8.0 *235 17.7 441 19.5 (d)403 35.3 456 22.8 241 20.5 233 11.7 (d) 390 34.7 Id)439 20.7 (d)423 20.0 258 19.8 *253 30.8 (b)49.0 2.46 (b) 42.0 2.52 (d) 41.1 1.59 (O39.0 1.68 45.7 2.13 (041.9 2.01 (d) 37,6 1.84 (d)40.3 2.12 (d) 43,3 1.76 lb) 47.5 3.48 *(c)38,6 2.03 Id) 37.2 4.24 (d) 37.9 2.45 51.0 4.94 (d) 45.3 3.63 NOT FOR DISTRIBUTION OR ATTRIBUTION SAL 000066926 1,2-Dichloroethane, NTPTOX 4, Draft 6/89 A-6 NOT FOR DISTRIBUTION OR ATTRIBUTION airtT c n o m c T o io iiT m ti TABLE AS. HEMATOLOGIC AND SERUM CHEMICAL DATA FOR MALE SPRAGUE DAWLEY RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF lJDICHLOROETHANE (Continued) Analysis Day Control 2.000 ppcn 4,000 ppm 8,000 ppm Blood urea nitrogen (mg/dl) Creatine kinase ilU/liter) Sorbitol dehydrogenase UU/liter) 3 7 14 45 90 3 7 14 45 90 3 7 14 45 90 (d) 21.7 0.75 18.6 1.74 lb) 23.4 1.07 L6.6 1.33 22.4 * 1.21 808 69 891 115 (d)742 80 829 t 45 818 73 9.9 (b)7.1 ' (d) 9.8 11.5 5.8 1.37 0.44 0.49 1.52 1.04 ib) 25.6 1.85 *<d) 28.4 3.56 (d> 26.3 1.22 "(d) 22.3 0.53 20.6 1.06 id) 21.2 1.74 (d) 26.3 2.82 (0 25.4 3.14 Id) 19.3 1.63 22.4 1.51 (d) 27.2 2.18 (b) 26.4 3.61 (b) 28.6 2.82 22.1 1.69 23.9 1.77 1,035 119 1,079 101 (d)889 72 1,220 234 1.098 98 1,008 > 82 989 112 1.057 127 1.026 100 959 98 <d) 1.186 163 id)898 69 Id)822 4* 49 863 98 739 46 (d) 10.1 1.09 (d) 7.6 0.24 10.9 1.04 9.2 i 0.36 7.0 0.78 8.7 0.40 (08.6 0.84 10.5 0.34 10.6 0.45 7,4 1.27 if) 11.0 1.35 id) 8.3 0.47 lb) 11.8 0.75 13.3 2.16 7.9 1.05 (a) Mean standard error for groups of 10 animals unless otherwise specified; P values vs. the controls by Dunn's test (Dunn, 1964) or Shirley's test (Shirley, 1977). (b) Eight animals were examined. (c) Seven animals were examined. (d) Nine animals were examined. (e) Six animals were examined. (f) Four animals were examined. *P<0.05 "PC0.01 < F c c t c C A-7 l, 2-Dichloroethane, NTP TOX 4, Draft 6/89 SAL 000066927 i TABLE A7. ABSOLUTE ORGAN WEIGHTS FOR OSBORNE-MENDEL RATS IN THE THIRTEEN-WEEK DRINKING WATER STUDIES OF l ,2-DlCHLORO ETHANE (a) Control 500 ppm 1,000 ppm 2,000 ppm -4.000 ppm 8,000 ppm MALE Number weighed (b) 9 10 10 10 10 Brain 2,056 33 2.106 25 2.089 36 1.995 37 1,991 57 Heart 1.498 74 1.526 48 1.605 70 1,386 62 1,289 53 Right kidney 1.506 36 1.600 41 *1.751 40 1,656 59 1.613 44 Liver 16.230 810 17.830 610 "21.080 840 19.310 800 15,190 510 Lung 1.821 80 (c) 1,946 113 2,075 61 2.074 123 1.960 83 Right testis 1,725 59 1.655 64 1,747 40 1.725 82 1.635 53 Thymus 314 19 305 28 323 19 314 22 326 22 10 1.982 so 1,295 59 1,507 68 15.900 800 1,717 53 1.631 38 333 27 FEMALE Number weighed 10 10 10 10 10 10 Brain 1.936 37 Heart 1,012 40 Right kidney 894 28 Liver 10.390 450 Lung 1,532 68 Thymus 304 14 1,996 26 1,051 28 *1.017 15 11.580 360 1,612 69 319 25 1.956 22 1,907 23 1,965 + 30 1,022 62 939 22 980 34 "1.041 22 "1,020 24 "1,096 37 10,810 230 10,390 430 10,750 300 1.629 81 1,497 79 1,565 57 278 40 309 34 341 25 1,933 33 909 22 "1,094 33 10,100 410 1,571 51 258 16 la) Mean standard error in milligram* unleu otherwise specified; P values vs. the control* by Dunn's test (Dunn, 1964) or Shirley's test (Shirley, 1977). (b) Unleu otherwise specified (c) Lungs of nine animal* were weighed. "P<0.01 NOT FOR DISTRIBUTION OR ATTRIBUTION 1,2- Dichloroethane, NTP TOX 4, Draft 6/89 A-8 CD C-i O' O <j O o o o -> <L CD $