Document zzV5qLVOJrNnzwan4jnvvM42B

AR226-3118 " | Li:iffenSecin, egncte}ns EE a ee | D Ce a e a onA Dosa e y L cL oe l l laad ii na oane RR 1 He asne 0 R d a ch o | i l FCaSel Sn Sr | E D aa Ge pie i Hh da i 1 PUR ' P fEEP He Boa dey Bil Hr i SCACB! mani o.T . |j )) haSnns YEdni el aii| : bnne alaR in 5coSh eoE a tells E Aa SAE EE E CONFIDENTIAL DPT439/984974 ACUTE TOXICTTY TO FISH Sponsor DuPont Speciality Chemicals, Jackson Laboratory, Chambers Works, Deepwater, NJ 08023, USA. Research Laboratory Huntingdon Life Sciences Ltd., Eye, Suffolk IP237PX, ENGLAND. Draft Report Issued 3 February 1999 Final Report Issued 22 March 1999 Page 1 of 27 Company Sanitized. Doss net contain TSCA OB) DPT439/984974 CONTENTS Page COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS.................... 3 QUALITY ASSURANCE STATEMENT............................................................................ 4 CONTRIBUTING SCIENTISTS.......................................................................................... 5 6 SUMMARY........................................................................................................................... INTRODUCTION............................................................................................................ 7 TEST SUBSTANCE............................................................................................................. 8 EXPERIMENTAL PROCEDURE........................................................................................ 9 MAINTENANCE OF RECORDS......................................................................................... 12 13 RESULTS.............................................................................................................................. CONCLUSIONS................................................................................................................... 14 REFERENCE...........................--.........--....--...--...........................--.............--........ 14 FIGURE 1. Typical sample chromatogram ...--..........--...--..--.............................;................... 15 TABLES 1. Measured concentrations...............-.............-..............--.....--.................................. 16 2. Sub-lethal effects .................--.............--....................--..-- --....--.................... 17 3. Environmental parameters ..........................................................--..--.................... 18 APPENDICES 1. Typical water quality characteristics of the diluent supply ........................................ 19 2. The determination o^ulHMIHRi^ aqueous media ............................................. 20 :2 : Company Sanitized. Doss not contain TSCA 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 Laboratory 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 of Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM(98)17. '-"..'.i Eileen C. Daly, Nat. bipfomaN, CEA Eire, Study Director, Huntingdon Life Sciences Ltd. 2 2 " ' ^ W i Mvr*~*f ^. Date ; :3 : Company Sa-'siiized. Doss net contain TSCA CB! 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 Fonnujatipnoftest medium Sampling of test medium Experimental set-up Report Audit 28 August 1998 09 September 1998 02 October 1998 02 October 1998 26 October 1998 24 February 1999 02 September 1998 09 September 1998 02.0ctober 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 of routine and repetitive procedures employed on this type of 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. t ^Mc^rc^jwoi Date 4 Company SsnEtized. Cess nb6 contain TSCACBE Company Sanil-??^ ^-c "'* c""^'" T?;^ cs; 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 Martin Nash, B.Sc.(Hons.) Scientific Officer Company Sanitized. Does nci contain TSCA CBl SUMMARY DPT439/984974 A study was performed to assess the acute toxicity ^(IHUHB10 "iobow trout (Oncorhynchus mykiss) under semi-static exposure conditions with 24-Eburly renewal of the test medium. Throughout the report, the exposure concentration and test results have been expressed in terms of the active The study was conducted in accordance with EEC Methods for Determination ofEcotoxicity 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". to|fUHBj A group of ten juvenile fish was exposed dissolved in water at a nominal concentration of 100 mg a.i./l; to aid dissolution, ultrasound treatment was employed. The measured concentrations of^D^^^fH^^H^J^H^rAanged between 91 and 94% of the nominal value in unfiltered samples of freshly prepared medium and between 101 and 109% of its starting value in samples of expired medium. The overall mean measured level olJUlUUMwas 94.1 mg a.i./l. At 96 hours, one fish exposed toIf^^^B^^^^H----^^^RmiAat 94.1 mg a.i./l had died. Sub-lethal effects comprising hyperventilation and ^^^^^^^^^^^^^y darkened pigmentation were also observed. The 96-hour LC,o was not identified but must be >94.1 mg a.i./l. taMBMBf^the Because sub-lethal effects were exhibited by fish exposed "no-observed effect concentration" (NOEC) was not identified in the limit test; however, based on the results of an earlier rangefinding 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 PreparationsgM^UUHis not considered to require classification as the 96-hour LCso is 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 Ssnitasd. Does r.ot contain TSCA GB? INTRODUCTION DPT439/984974 This study was designed to assess the acute toxicity ofjteBBH^|jto rainbow trout (Oncorhynchus mykiss) under semi-static exposure conditions. The study was conducted in accordance with EEC Methods for Determination ofEcotoxicity 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 of chemical analysis were issued by 12 October 1998. ofBHHfjui Information provided by the Sponsor indicated that the solubility water wasH^I 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 wanned to 35 - 40C (in a water bath) with gentle stirring to obtain a homogenous composition before use. :7 : Comply SanitEzed. Do.s rot co^;n TSCA CSt Identity: Chemical name: Appearance: Storage conditions: Lot number: Expiry date: Purity: Sample received: TEST SUBSTANCE 23 June 1998 DPT439/984974 :8 : Company S^ anitize^ d. nDo-s rro.jt^contain TSCA CBl 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. Acclimatisation The stock of fish was obtained from the supplier on 28 July 1998 and they were held in an aerated supply of 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 CaCO,. The fish were fed daily with commercial fish food (TROUW (UK) Ltd; Nutra Fry 02) an amount equivalent to 2% of the total wet-weight offish 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 EIga 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 Sanitised. Does r:c-t contain TSCA CBl DPT439/984974 TEST SUBSTANCE PREPARATION Method of 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 of the test concentration o^uiimH^as The test concentration measured using an HPLC method of chemical analysis (Appendix 2). The stability of the test substance hi dechlorinated tap water was determined under storage conditions of4C and room temperature before the start of the study (Table 2, Appendix 2). Four, mid-vessel samples (100 ml) of 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 of the samples from each set were transferred to the Huntingdon Research Centre of 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 of 0.99 g bodyweight/litre. Test concentrations The rangefinding study was conducted with test concentrations of 1, 10 and 100 mg/1. Based on the results of this test, the definitive (limit) test employed a nominal concentration of 100 mg/1. Medium renewal The fish were exposed to the control or test conditions for a period of 96 hours with daily batahwise renewal of the media to ensure the maintenance of satisfactory environmental conditions and to maintain a stable exposure level. 10 : Company Sait^d. Do-3 notc^n TSCAC31 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 of 16 hours light: 8 hours dark was maintained, with periods of subdued lighting at the beginning and end of each light phase. Daily records of 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 of death employed in this study were (i) absence of respiratory movement and (ii) absence of response to physical stimulation of 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 of 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 Compsny Senitized. Doss not contain TSCA CB's' DPT439/984974 MAINTENANCE OF RECORDS All specimens, raw data and study related documents generated during the course of 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 of 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. : 12 : Company S3..i-. ^ nct cont31n T5C& CB5 DPT439/984974 RESULTS Chemical analysis The results of chemical analysis are given in Table 1 and an example chromatogram is illustrated in ~ ' Concentrations are expressed in terms of the measured levels of the active ingredie Results for unfiltered samples offfffffffyndicated that the intended exposure concentration was adequately achieved (between 91 and 100% of the nominal value) and maintained during the test (between 101 and 109% of the starting value). The overall mean measured concentration o^H----| ----Kvas 94.1 mg a.i./l. Mortality and observations One fish had died (10% mortality) at 94.1 mg a.i./l at 96 hours. The 96-hour U^o could not be calculated but must be >94.1 mg a.i./l. A chronological record of 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,!------l^s not considered to require classification as the 96-hour LCso is considered to be greater than the highest nominal concentration, 100 mg a.i./l (mean measured level of 94.1 mg a.i./l). Environmental parameters The measurements of water quality (temperature, pH, concentrations of 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 : Compan;; Sanitized. Does not conta'n TSCA C3; CONCLUSIONS DPT439/984974 Iwas not found to be toxic to rainbow treat when dissolved in water at 94.1 mg active | "ingredient/I. The "no-observed-effect concentration", based on the results of the rangefinding test, was considered to be 1 mg a.L/1 (nominal concentration). REFERENCE ./ Official Journal of the European Communities Commission Directive (1 March 1991). Annex VI | General classification and labelling requirements for dangerous substances and preparations. Part n "Classification on the Basis a/Environmental Effect" p.62 - 64. : 14 : Company Sanitized. Dozs 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 c INJECT 09-10-98 18:2S:47 STORED TO BIN 116 Company Sanitized. Does not coritaiii TSGA CBS TABLE 1 Measured concentrations DPT439/984974 Nominal cone.,' mg/1 0 hours 0 nd nd 100 91.2 91.7 [92] Measuredj|JMlB|Qponcentratioinngs/,1 24 hours '^^itiT 72 hours 96 hours nd nd - nd nd nd nd 92.5 99.8 105 90.8 93.7 98.4 94.4 [96] [92] [96] Overall %ti mean - - 104 94.1 (94) nd % ti [ ] ( ) none detected (< 2.5 mg a.i./l). mean measured concentration after 24 or 96 hours expressed as a percentage of the mean starting concentration (0 and 72 hours). mean measured concentration expressed as a percentage of me nominal concentration. overall mean measured concentration expressed as a percentage of the nominal concentration. : 16 : Company Ssr.Etized. Does not cor.Sain TSCA CB1 TABLE 2 Sub-lethal effects DPT439/984974 (n g/1) nominal1 measured1 ' Abnormality Control nd Aggression Aggregation 100 94.1 Aggression Aggregation Darkened pigmentation (eye orbit only) Darkened pigmentation (eye and body) Hyperventilation Initial populatiori=10fis h/conoentrarion 025h 2h 4h 24 h 48 h 72 h 96 h - - - 1/10 - 1/10 - 9/10 9/10 1/10 - - 8/10 - 1/10 3/10 1/9 1/10 3/10 3/9 10/10 9/9 x/y number offish affected / number offish surviving. nd none detected (< 2.5 mg a.L/1). i : 17 : Company Sanitized. 3oes not contain TSCA CBi TABLE 3 Environmental parameters temperature, pH, dissolved oxygen and total hardness DPT439/984974 T T (ing/l) nominal* measured1 TemperatureC mm max. Control nd 14.0 15.1 100 94.1 14.1 15.2 P H min max 7.6 7.9 7.3 7.8 Dissolve doxygen (%AISV) min max 81 103 79 103 Total hardness mg/I as CaC03 0 hours 202 204 s ASV nd as.active-ingredie air saturation value.' none detected (<2.5 mg a.i71). Continuous monitoring of control vessel media temperature = 13.8 to 15.3 C. : 18 : Company Sanitized, uoes noS contain TSCA CBt 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 CaCO>3. Analysis in March 1998 gave the following results: Colony count 37C Colifonn organisms MPN per 100 ml E. coli type 1 MPN per 100 ml Organochlorine pesticides Polychlorinated biphenyls Qrganpjihosphorus pesticides Aluminium Ammonia Arsenic Boron Cadmium Calcium Carbon (tota! organic) Chloride Chlorine (free) Chlorine (total) Chromium C.O.D. Cobalt Copper Fluoride Iron Lead Magnesium Mercury Nitrate (as N03) Nitrite (as NOz) Nickel Phosphorus Potassium Silver Sodium Tin Total suspended solids Zinc conductivity turbidity 0 nil nil us/I 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 |iS/cm 0.16FTU MPN most probable number nd not determined. 19 : Company Sam^zed. Dees not contain TSCA CBl APPENDIX 2 THE DETERMINATION 0 DPT439/984974 AQUEOUS MEDIA SAMPLE ANALYSIS The aqueous samples were diluted with sodium hydroxide and acetonitrile to bring the ex ofnimURwas Hl^lHBRconcentrations within the calibration range. Determination by ugh performance liquid chromatography (HPLC) using a conductivity detector1. 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: Dimensions (1 x id): Temperature: Mobile phase Composition: Flow rate: Suppressor type: Regenerant composition: Flow rate: Injection volume: PLRP-S supplied by Polymer Laboratories 250 x 4.6 mm Ambient Acetonitrile : aqueous buffer solution (25 : 75% v/v) l.Oml/min 50 mN sulphuric acid 2.5 ml/min 100 pi Aqueous buffer solution: 2mM ammonium hydroxide/ImM 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). ' 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 : Compeny San^sac'. Dcss nat contain TSCA CBE DPT439/984974 CALIBRATION SOLUTIONS Calibration solutions were prepared with the same batch ofJii^lMBlused 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./I) 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./I) 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. CALCULATIONS UBHlHB|poncentrations were determined using mean bracketing standards. ^nHI^^Utt The mean peak height responses were calculated m bracketing standard chromatograms. ThenlBH^U^oncentration in each sample was then calculated using the following equation: ^ . Where Y = Z = A = F = tc^UUHM Detector response Mean detector response to bracketing standard. Concentration of bracketing standard (mg/I). Factor to take into account sample processing. The purity (active ingredient) of the test substance isjriven in the Test Substance Data Sheet asgBU asj|f|^(as Nominal and fortified concentrations are reported asnil------as supplied and in terms ofthe active ingredient. VALIDATION OF THE ANALYTICAL PROCEDURE The analytical procedure was validated by determining the linearity of response of the analytical system, specificity of chromatographic analysis, the limit of detection, and the method's accuracy and precision. During the course ofthe study the performance ofthe method was monitored by the analysis of quality control samples. : 21 : Compsny Sanlteaa. uOafi nut contsiu fSCA CBE Validation recoveries o: DPT439/984974 TABLE 1 from fortified samples of dilution media Medium Dechlorinated tap water Algal Elendt M4 Overall mean (RSD) Fortification level (mg/1) As supplied Control 387.9 387.9 Control 535.5 530.0 214.2 212.0 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 % of 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) RSD: relative standard deviation. ND: none detected; less than the limit of detection (trout and algal studies : 2.5 mg a.L/1; Daphnia study: 0.5 mg a.i./l). The limit of 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 CBc Stability of TABLE 2 [in dechlorinated tap water DPT439/984974 Storage conditions Procedural recovery' Procedural recovery' Light, sealed, room temperature' Light, sealed, room temperature ' Dark, sealed, 4C' Dark, sealed, 4C' Procedural recovery2 Procedural recovery2 ' Fortification level: 11.20 mg/1 fortification level: 11.65 mg/1 Results are given as percentage recoveries o: Time-point 0 hours 20 hours 110 - 112 - - 103 - 106 - 109 - 93.1 - 109 - 98.9 |hfter storage for the indicated time. : 23 Company Sanitizod. Does not contam TSCA CSt o ? w L/> ^ (s) >- ^. ^. i*> >-- o w o " ? S " - y ^ b ~~1 -~) 0 \0 ~J -- B ^ 4^. m a ^ S >s) >^ ^ 3 o ^ %^ ^ ?^. g- '& Response FIGURE! Standard Calibration foi^j (ca 4 to 50 mg/1) DPT439/984974 Runno.:DPT/440/002 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 CBi DPT439/984974 FIGURES 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" ' L rNJECT' 09-07-78' T4:55;T7" STORED TtrBTmt 8'5 DATA SAVED TO BIN 8 -85 : 26 : Company 5S^^u^sde.aD. oe3.=tc^nTSCACSl DPT439/984974 FIGURES Typical chromatography Sample of algal medium fortified at 535.5 mg/I (= 133.9 mg/1 as a.i.) CHANNEL A INJECT 09-09-9813:19:59 STORED TO BIN 75 6.36 .! DATA SAVED TO BIN 75 : 27 : Company Sanitized. Does r.oS contain TSCA CD?