Document NE9MRegzyKymv5Mn7vr0Z9XDD

untmg fi' Gfj=*-. -rTLL-A li s*c S r ip nKiiaca*fHai t a lVV'-gMTffV`r."TtSf?i- ~\ \ r,ia> m S C A C M Co3s notco' COiTPan^ San*** . ;.j . 3-*'Vii :; U t' CONFIDENTIAL DPT439/984974 ACUTE TOXICITY TO FISH Sponsor DuPont Speciality Chemicals, Jackson Laboratory, Chambers Works, Deepwater, NJ 08023, USA. Research Laboratory Huntingdon Life Sciences Ltd., Eye, Suffolk IP23 7PX, ENGLAND. Draft Report Issued 3 February 1999 Final Report Issued 22 March 1999 Page 1 o f 27 Company Sanitized. Doss r.ct coni, TSCA CBf CONTENTS COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS QUALITY ASSURANCE STATEMENT........................................................ CONTRIBUTING SCIENTISTS...................................................................... SUMMARY.............................................. ........................................................ INTRODUCTION......................................................... .................................... TEST SUBSTANCE.............................-............................................................ EXPERIMENTAL PROCEDURE..................................................................... MAINTENANCE OF RECORDS...................................................................... RESULTS.......................... ................................................................................. CONCLUSIONS................................................................................................. REFERENCE...................................................................................................... DPT439/984974 Page 3 4 5 6 7 8 9 12 13 14 14 FIGURE 1. T yp ical sam ple ch rom atogram ............................................................................. 15 TABLES 1. M easured concentrations........................................................................................ 2. Sub-lethal effects..................................................................................... 3. Environmental parameters ...................................................................... 16 17 18 APPENDICES 1. Typical water quality characteristics of the diluent supply..................... 2. The determination a9ueous niedia......................... :2 : 19 20 Company Sanitized. Doss not contain TSG CEE DPT439/984974 COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS The study described in this report was conducted in compliance with the following Good Laboratory Practice Standards and I consider the data generated to be valid. The UK Good Laboratoiy Practice Regulations 1997 (Statutory Instrument No. 654). EC Council Directive 87/18/EEC of 18 December 1986 (Official Journal No. L 15/29). OECD Principles o f Good Laboratoiy Practice (as revised in 1997), ENV/MC/CHEM(98)17. Study Director, Huntingdon Life Sciences Ltd. Date :3 : Company Sanitized. Dass net contain T3CA CBI QUALITY ASSURANCE STATEMENT The following have been inspected or audited in relation to this study DPT439/984974 Study Phases Inspected Protocol Audit Date of Inspection 04 August 1998 Date of Reporting 04 August 1998 Process Based Inspections Fish observations Test medium renewal Formulation of test medium Sampling of test medium Experimental set-up Report Audit 28 August 1998 09 September 1998 02 October 1998 2 October 1998 26 October 1998 24 February 1999 02 September 1998 09 September 1998 02_October 1998 02 October 1998 26 October 1998 24 February 1999 Protocol Audit: An audit of the protocol for this study was conducted and reported to the Study Director and Company Management as indicated above. Process based inspections: At or about the time this study was in progress inspections o f routine and repetitive procedures employed on this type o f study were carried out. These were conducted and reported to appropriate Company Management as indicated above. Report Audit: This report has been audited by the Quality Assurance Department. This audit was conducted and reported to the Study Director and Company Management as indicated above. The methods, procedures and observations were found to be accurately described and the reported results to reflect the raw data. Helen Comb, B.Sc.(Hons.), Principal Auditor, Department of Quality Assurance, Huntingdon Life Sciences Ltd. Date 1 ; Company Sanitized. Ocas n! contain TSCCB1 Company SanithmA co n tai TP''* W CONTRIBUTING SCIENTISTS STUDY MANAGEMENT Eileen C. Daly, Nat. Diploma, NCEA Eire Study Director Rosalyn Mazey, B.Sc.(Hons.) Study Scientist Philip S. Manson, B.Sc.(Hons.), M.Sc. Study Scientist Ben Smith, B.Sc.(Hons.), M.Sc., C.Chem., M.R.S.C. Chief Chemist Andrew Robertson, B.Sc.(Hons.) Senior Chemist Richard Cubberley, B.Sc.(Hons.) Study Analyst I Martin Nash, B.Sc.(Hons.) Scientific Officer DPT439/984974 :5 : Company ;[n TSCAOB! SUMMARY DPT439/984974 A study was performed to assess the acute toxicity rainbow trout (Oncorhynchus mykiss) under semi-static exposure conditions with 24-Eourly renewal o f the test medium. Throughout the report, the exposure concentration and test results have been expressed in terms of the active ingredient (a.i/ The study was conducted in accordance with EEC Methods for Determination o f Ecotoxicity Annex to Directive 92/69/EEC (O.J. No. L383A, 29.12.92) Part C, Method 1 "Acute Toxicity for Fish" and the OECD Guideline for Testing of Chemicals No. 203 "Fish, Acute Toxicity Test". A group of ten juvenile fish was exposed t o | p B H ( l dissolved in water at a nominal concentration of 100 mg a.iJl; to aid dissolution, ultrasound treatment was employed. The measured concentrations o f { J ^ 0 H H P raned between 91 and 94% o f the nominal value in unfiltered samples o f freshly prepared medium and between 101 and 109% ^of its starting value in samples of expired medium. The overall mean measured level o n H H H H H Q w a s 94.1 mg a.i./I. At 96 hours, one fish exposed t o & m B H n at 94.1 mg a.i./l had died. Sub-lethal effects comprising hyperventilation and darkened pigmentaon were also observed. The 96-hour LC50was not identified but must be >94.1 mg a.iA Because sub-lethal effects were exhibited by fish exposed t q ^ H W H j t h e "no-observed effect concentration" (NOEC) was not identified in the limit test; however, based on the results of an earlier rangefmding test, the NOEC was considered to be 1 mg a.i/1 (nominal concentration). Under the EC General Classification and Labelling Requirements for Dangerous Substances and P r e p a r a tio n s A f l^ I B H H is not considered to require classification as the 96-hour LCS0is considered to be greater than the highest nominal concentration, 100 mg a.i./l (mean measured level of 94.1 mg a.i./l). :6 : Company Sanitizad. Doss not contain TSCA CB INTRODUCTION DPT439/984974 This study was designed to assess the acute toxicity o i j ^ H H I ^ j j t o rainbow trout (Oncorhynchus mykiss) under semi-static exposure conditions. The study was conducted in accordance with EEC Methods for Determination of Ecotoxicity Annex to Directive 92/69/EEC (O J. No. L383A, 29.12.92) Part C, Method 1 "Acute Toxicity for Fish" and the OECD Guideline for Testing of Chemicals No. 203 "Fish, Acute Toxicity Test". The protocol was approved by Huntingdon Life Sciences Management on 7 July 1998, by the Sponsor on 17 July 1998, and by the Study Director on 3 August 1998. The experimental phase of the study was conducted between 28 September and 9 October 1998 and the results o f chemical analysis were issued by 12 October 1998. Information provided by the Sponsor indicated that the solubility water w a s |p f ^ by weight at 35 - 40C and its purity was 25%. Throughout this report, the exposure concentration and test results have been expressed in terms of the active ingredient. Also, the Sponsor indicated that at room temperature the test substance was a suspension in water and upon standing it would separate out into its component phases; accordingly, at the recommendation of the Sponsor, the suspension was warmed to 35 - 40C (in a water bath) with gentle stirring to obtain a homogenous composition before use. :7 Company Sanitized. Do'.s not contain TSCA CS5 Identity: Chemical name: Appearance: Storage conditions: Lot number: Expiry date: Purity: Sample received: TEST SUBSTANCE DPT439/984974 Room temperature 2 years from date of receipt 23 June 1998 :8 : Company Sanitized. Do-s not contain TSCACBl EXPERIMENTAL PROCEDURE DPT439/984974 TEST SPECIES Name Rainbow trout (Oncorhynchus mykiss). Source - The fish were supplied by Parkwood Trout Farm, Kent, UK. They were reared at Bowerchalk Trout Farm, Salisbury, UK from South African eggs which hatched in June 1998. A cclim atisation The stock o f fish was obtained from the supplier on 28 July 1998 and they were held in an aerated supply o f diluent water under flow-through conditions until use. During the 14-day period immediately before the definitive test, temperatures remained within the range 13.6 to 13.9C, pH values within the range 7.8 to 7.9, dissolved oxygen concentrations within the range 98 to 100% air saturation value (ASV) and total hardness within the range 194 to 200 mg/1 as CaC03. Hie fish were fed daily with commercial fish food (TROUW (UK) Ltd; Nutra Fry 02) an amount equivalent to 2% o f the total wet-weight of fish in the holding tank. No food was given during the 27hour period immediately before exposure or during the exposure period itself. No medication was given during the holding period, and mortalities were recorded as 3% in the 14 days before the definitive test. The size of the fish used in the definitive study was determined by weighing and measuring a sample of ten fish taken at random from the holding tank on 29 September 1998; their mean fork length was 5.4 cm and their mean wet weight was 1.6 g. DILUENT WATER The water used to hold the fish and for the study was laboratory tap water, dechlorinated and softened by passage through an Elga water purification system. It was passed through a high grade activated carbon filter to remove chlorine and any organic contaminants. A proportion of the supply then passed through a water softener before final reverse osmosis treatment to produce a highly purified water supply. The two grades of dechlorinated water were then remixed to give a supply with the desired water hardness. This water was then held in an intermediate tank where it was equilibrated to the test temperature and gently aerated before being supplied to the holding and test areas. Typical water quality characteristics of the diluent supply are given in Appendix 1. :9 : Company Sanitized. Doss not contain TSCA CBl TEST SUBSTANCE PREPARATION DPT439/984974 Method o f preparation Based on information provided by the Sponsor, the test substance was wanned (to c.38C ) in a water bath and gently swirled to ensure a homgeneous mixture before weighing. The test substance (6.4 g) was mixed with diluent water (250 ml) before being poured into a volumetric flask (2 1) containing diluent water (1.5 1). The contents of the flask were treated by ultrasound for twenty minutes before being poured into a test vessel (glass aquarium). The volume was then adjusted to 16 litres with diluent water. Stability o f the test concentration The test concentration o f j j l H m ^ w a s measured using an HPLC method o f chemical analysis (Appendix 2). The stability of the test substance in dechlorinated tap water was determined under storage conditions o f 4C and room temperature before the start o f the study (Table 2, Appendix 2). Four, mid-vessel samples (100 ml) o f medium were taken from the control and test vessels at 0 and 72 hours (fresh media) and at 24 and 96 hours (expired media) and reserved for analysis. They were stored in a refrigerator before two o f the samples from each set were transferred to the Huntingdon Research Centre o f Huntingdon Life Sciences Ltd, Cambridgeshire, for analysis; the other two samples remained in storage at the Eye Research Centre in case further analysis was required. EXPOSURE CONDITIONS Experimental design A rangefinding test was followed by a definitive (limit) test with one test concentration plus one diluent water control. In the definitive test, ten fish were placed at random into each glass aquarium containing the prepared test or control media. Each vessel contained 16 litres of medium to a depth of 16 cm. This provided an initial static loading o f 0.99 g bodyweight/litre. Test concentrations The rangefinding study was conducted with test concentrations o f 1, 10 and 100 mg/1. Based on the results of this test, the definitive (limit) test employed a nominal concentration o f 100 mg/1. Medium renewal The fish were exposed to the control or test conditions for a period of 96 hours with daily batchwise renewal of the media to ensure the maintenance of satisfactory environmental conditions and to maintain a stable exposure level. 10 Company Sanitised. Do-3 contain TSCA CSt DPT439/984974 Environmental conditions Treatment and control groups were maintained at 15 2C throughout the exposure period and constant to within 1C during the study. The temperature of the water in the control vessel was continuously monitored during the study. Supplementary aeration was provided via narrow bore glass tubes. A photoperiod o f 16 hours lig h t: 8 hours dark was maintained, with periods of subdued lighting at the beginning and end of each light phase. Daily records o f temperature, pH and dissolved oxygen were kept for each control and test vessel together with measurements of total hardness for selected vessels at 0 hours. The fish were not fed during the 96 hour exposure period. CRITERIA OF EFFECT The criteria o f death employed in this study were (i) absence of respiratory movement and (ii) absence o f response to physical stimulation o f the caudal peduncle. In addition to observations on mortality at 15 minutes, 2, 4, 24, 48, 72 and 96 hours, subjective assessments were also made on the incidence and type o f any sub-lethal effects compared with control fish. EVALUATION OF DATA The "no-observed-effect concentration" (NOEC) was derived by direct inspection of the data on the treatment-related-effects. An incidence rate of more than one affected fish out of ten is considered to be significant. PROTOCOL DEVIATIONS None. : 11 : Company Sanitized. Does not contain TSCA CE DPT439/984974 MAINTENANCE OF RECORDS All specimens, raw data and study related documents generated during the course o f the study at Huntingdon Life Sciences, together with a copy of the final report will be lodged in the Huntingdon Life Sciences Archive. Such specimens and records will be retained for a minimum period of five years from the date of issue o f the final report. At the end of the five year retention period the Sponsor will be contacted and advice sought on the future requirements. Under no circumstances will any item be discarded without the Sponsor's knowledge. \ i l: I 1 : 12 : Hzed. Does not contain TSCA C&> DPT439/984974 RESULTS Chemical analysis The results of chemical analysis are given in Table 1 and an example chromatogram is illustrated in Figure 1. Concentrations are expressed in terms of the measured levels of the active ingredient, Results for unfiltered samples o fJ ^ m ^ ^ n d i c a t e d that the intended exposure concentration was adequately achieved (between 91 and 100% of the nominal value) and maintained duringJ h e test (between 101 and 109% of the starting value). The overall mean measured concentration o | ^ P ^ v a s 94.1 mg a.i./l. M ortality and observations One fish had died (10% mortality) at 94.1 mg a.i./l at 96 hours. _____ The 96-hour LC50could not be calculated but must be >94.1 mg a.i./l. A chronological record o f sub-lethal effects is given in Table 2. They comprised hyperventilation, darkened pigmentation, aggression and aggregation. At 24 and 48 hours, nine fish and one fish were affected respectively, and at 72 hours all of the fish were affected. At 96 hours, when one fish had died, the remaining fish were adversely affected although their symptoms may have been caused by a fish which showed aggressive behaviour. Aggregation was also exhibited by nine fish in the control group at 24 and 72 hours and is also attributed to the presence of an aggressive fish. The "no-observed-effect" concentration (NOEC) could not be identified in the limit test; based on the rangefinding test, the NOEC was considered to be 1 mg a.i./l. (nominal concentration). Under the EC General Classification and Labelling Requirements for Dangerous Substances and Preparations,flH B H H H pns not considered to require classification as the 96-hour LCS0is considered to be greater tm!n the highest nominal concentration, 100 mg a.i./l (mean measured level of 94.1 mg a.i./I). Environmental parameters The measurements o f water quality (temperature, pH, concentrations o f dissolved oxygen and total hardness) are summarised in Table 3; they remained within acceptable limits throughout the study. The test medium was clear and colourless. : 13 : Company iized. Does not contain T5CA CB. CONCLUSIONS DPT439/984974 was not found to be toxic to rainbow trout when dissolved in water at 94.1 mg active ingredient/1. The "no-observed-effect concentration", based on the results o f the rangefinding test, was considered to be 1 mg a.i./l (nominal concentration). REFERENCE Official Journal o f the European Communities Commission Directive (1 March 1991). Annex VI General classification and labelling requirements for dangerous substances and preparations. Part II "Classification on the Basis o fEnvironmental Effect" p.62 - 64. w : 14 Company Sasnttized. Doss not contain TSCA CBi DPT439/984974 FIGURE 1 Typical sample chromatogram - 400 mg/1 (100 mg a.i./I) taken on Day 1 of the test CHANNEL A INJECT 09-10-98 18:25:47 STORED TO BIN # 116 : 15 : Company Sanitized. Does not contain TSCA CBS TABLE 1 Measured concentrations DPT439/984974 Nominal cone.,* mg/1 0 hours 0 nd nd 100 91.2 91.7 [92] Measured 24 hours nd nd 92.5 99.8 [96] Rponcentrations, i ng/1 %ti 72 hours 96 hours - nd nd 105 90.8 93.7 [92] nd nd 98.4 94.4 [96] Overall % ti mean -104 94.1 (94) s in terms o f die active ingredient, nd none detected (< 2.5 mg a.i./l). % ti mean measured concentration after 24 or 96 hours expressed as a percentage o f the mean starting concentration (0 and 72 hours). [ ] mean measured concentration expressed as a percentage o f die nominal concentration, ( ) overall mean measured concentration expressed as a percentage o f the nominal concentration. : 16 : Company Sanitized. Does not contain TSCA CBl TABLE 2 Sab-lethal effects DPT439/984974 (mg/1) nominal* measured* ' Abnormality Control nd Aggression Aggregation Initial population = 1 0 fish / concentration 0.25h 2 h 4 h 24 h 48 h 72 h 96 h . 1/10 . 1/10 - - - 9/10 - 9/10 - 100 94.1 Aggression Aggregation . Darkened pigmentation (eye orbit only) Darkened pigmentation (eye and body) Hyperventilation _ _ _ 1/10 - - - 8/10 - - - - - 1/10 - 3/10 1/9 - - - - 1/10 3/10 3/9 - - - - - 10/10 9/9 x/y number of fish affected / number of fish surviving. nd none detected (< 2.5 mg a.i./l). s as active in g re d ie n t,||B H B H I^ H H V l : 17 : Company Sanitized. Soei not contain TSC C'Bt TABLE 3 Environmental parameters temperature, pH, dissolved oxygen and total hardness DPT439/984974 m m n r TemperatureC (nig/i) nominal1 measured1 min m ax. Control nd 14.0 15.1 100 94.1 14.1 15.2 pH min max 7.6 7.9 7.3 7.8 Dissolved oxygen ( % /lSV) min max Total hardness mg/1 as CaC03 0 hours 81 103 202 79 103 204 * ASV nd as active-ingredien air saturation value. none detected (<2.5 mg a.i71). Continuous monitoring o f control vessel media temperature = 13.8 to 15.3 C. : 18 : Company Sanitized- uoes not r.tain go TSCA CBl DPT439/984974 APPENDIX 1 TYPICAL WATER QUALITY CHARACTERISTICS OF THE DILUENT SUPPLY Diluent water comprised filtered dechlorinated tap water blended with tap water that had been softened and subsequently treated by reverse osmosis to a hardness of approx. 200 mg/1 as CaC03. Analysis in March 1998 gave the following results: Colony count 37C Coliform organisms MPN per 100 ml E. coli type 1 MPN per 100 ml Organochlorine pesticides Polychlorinated biphenyls Organophosphorus pesticides Aluminium Ammonia Arsenic Boron Cadmium Calcium Carbon (total organic) Chloride Chlorine (free) Chlorine (total) Chromium C.O.D. Cohalt Copper Fluoride Iron Lead Magnesium Mercury Nitrate (as NO3) Nitrite (as NO2) Nickel Phosphorus Potassium Silver Sodium Tin Total suspended solids Zinc conductivity turbidity 0 nil nil ixg/1 <0.02 <0.02 <0.03 mg/1 <0.01 <0.03 <0.0015 <0.05 <0.0004 76 1 25 0.06 0.08 <0.001 <15 <0.001 0.02 0.2 <0.01 <0.0019 4.1 <0.0001 6.46 <0.01 0.003 <0.2 1.78 <0.001 13.4 nd <2 0.0125 419 pS/cm <0.16 FTU MPN most probable number nd not determined. : 19 : Company Sanitized. Does not contain TSCA CBS APPENDIX 2 THE DETERM INATION OF DPT439/984974 AQUEOUS M EDIA SAMPLE ANALYSIS i T_h_e__ ueous samples were diluted with sodium hydroxide and acetonitrile to ^ jjH iA c o n c e n tr a tio n s within the calibration range. Determination igh performance liquid chromatography (HPLC) using a conductivity detector ;ted Lwas by CHROMATOGRAPHY INSTRUMENTATION AND CONDITIONS A high performance liquid chromatography system comprising autosampler, pump, conductivity detector, anion suppressor and data collection system was used. Column Type: i Dimensions (1 x id): PLRP-S supplied by Polymer Laboratories 250 x 4.6 mm Temperature: Ambient Mobile phase Composition: Acetonitrile : aqueous buffer solution (25 : 75% v/v) Flow rate: 1.0 ml/min Suppresser 1ype: Regenerant composition: 50 mN sulphuric acid Flow rate: 2.5 ml/min * Injection volume: 100 p.1 Aqueous buffer solution: 2mM ammonium hydroxide/lmM sodium carbonate (made up in ultra high purity water) and filtered through 0.2 micron cellulose nitrate filter paper. Under the above condition hromatographed as a single peak (see Figure 3). I ' A method contained in a fax dated 23 July 1998 from Kavsy D Dastur, Dupont Specialty Chemicals was modified to comply with Huntingdon Life Sciences standard operating procedures and instrumentation. : 20 : Company Sanifod. Dess not contain TSCA CB5 DPT439/984974 CALIBRATION SOLUTIONS Calibration solutions were prepared with the same batch o f | ^ m ^ J u s e d in the preparation of the toxicity test solutions. Trout and Algae Studies Working calibration solutions in the nominal range 40 to 500 mg/1 (equal to 10 to 125 mg a.i./l) were prepared by volumetric dilution with acetonitrile: 100 mM sodium hydroxide (25:75% v/v) of a primary standard prepared in ultra high purity water. Daphnia Study Working calibration solutions in the nominal range 4.0 to 50.0 mg/1 (equal to 1.0 to 12.5 mg a.i./l) were prepared by volumetric dilution with acetonitrile: 100 mM sodium hydroxide (25: 75% v/v) o f a primary standard prepared in ultra high purity water. CALCULATIONS ncentrations were determined using mean bracketing standards. The mean peak heij chromatograms. The following equation: responses were calculated for in bracketing standard boncentration in eaci sample was then calculated using the Concentration .mg/1MV) = --Y x- --A x F,, Where Y Z A F Detector response to Mean detector response to bracketing standard. Concentration o f bracketing standard (mg/1). Factor to take into account sample processing. The purity (active ingredient) of the test substance is ;iven in the Test Substance Data Sheet Nominal and fortified concentrations are reported asi las supplied and in terms active ingredient. VALIDATION OF THE ANALYTICAL PROCEDURE The analytical procedure was validated by determining the linearity of response of the analytical system, specificity o f chromatographic analysis, the limit of detection, and the method's accuracy and precision. During the course of the study the performance o f the method was monitored by the analysis of quality control samples. : 21 : Company Sanitised. uOeS ut contain foCA CBI Validation recoveries o: DPT439/984974 TABLE 1 from fortified samples o f dilution media Medium Dechlorinated tap water Algal Elendt M4 Overall mean (RSD) Fortification level (mg/1) As supplied Control 387.9 387.9 Control 535.5 53. 214.2 212.0 Control 9.93 9.93 496.5 496.5 As active ingredient Control 96.98 96.98 Control 133.9 132.5 53.55 53.00 Control 2.483 2.483 124.1 124.1 Recovery as a % o f fortification level ND 95.8 104 ND 92.0 101 92.6 95.5 ND 98.5 95.9 103 103 98.1 (4.2) Active ingredient content of RSD: relative standard deviation. ND: none detected; less than the limit o f detection (trout and algal studies : 2.5 mg a.i./l; D aphnia study: 0.5 mg a.i./l). The limit o f detection is defined as the analyte concentration in a processed sample which would give a peak equal to 3 x local base-line noise. : 22 : Company Sanitized. Does not contain TSCA CBs DPT439/984974 TABLE 2 Stability o: in dechlorinated tap water Storage conditions Procedural recovery1 Procedural recovery1 Light, sealed, room temperature' Light, sealed, room temperature 1 Dark, sealed, 4 C 1 Dark, sealed, 4C 1 Procedural recovery2 Procedural recovery2 Time-point 0 hours 20 hours 110 112 - - 103 - 106 - 109 - 93.1 - 109 - 98.9 1Fortification level: 11.20 mg/1 2Fortification level: 11.65 mg/1 Results are given as percentage recoveries oi j^ifte:r storage for the indicated time. : 23 : Company Sanitized. Does not contain TSCA CBt FIGURE 1 Standard Calibration for (cfl 40 - 500 mg/1) DPT439/984974 Concentration (m Run no.:DPT/439/013 Standard concentration (mg/1) 0.0 43.57 108.9 217.9 435.7 544.7 NOP: no observable peak. l) Peak height NOP 4075 10668 20734 38644 47864 Company Sanitized. Does not contain TSCA CBI FIGURE 2 Standard Calibration for (ica 4 to 50 mg/I) 0 DPT439/984974 Run no.:DPT/44Q/002 Standard concentration (mg/1) 0.0 4.720 11.80 23.60 47.20 59.00 NOP: no observable peak. Peak height NOP 3295 8657 18669 38224 48699 : 25 : Company Sanitised. Does not contain TSCA CB1 DPT439/984974 FIGURE 3 Typical calibration chromatography - 217.9 mg/1 (= 54.47 mg/1 as a.i.) CHANNEL A INJECT 09-09-98 14:45:47 STORED TO BIN ft 84 DATA SAVED TO BIN ft 84 FIGURE 4 Typical chromatography - Unfortified algal medium CHANNEL A " TNJECT 09-09-98' 14:55 : r r STORED TtrBTTr f t 85 { DATA SAVED TO BIN ft '85 26 Gonisin TSCA C il Company S e i z e d - Does : DPT439/984974 FIGURE 5 Typical chromatography Sample of algal medium fortified at 535.5 mg/1 (= 133.9 mg/1 as a.i.) CHANNEL A rINJECT 09-09-9813:19:59 STORED TO BIN # 75 DATA SAVED TO BIN # 75 : 27 : Company Sanitized. Does not contain TSCA Ciii