## REPORT

Acute ToxicityTo Rainbow Trout

## (Oncorhynchus mykiss)

DataRequirements/Test Guidelines: • OECD Guidelines for Testing ofChemicals (1992) No 203, "Fish, Acute Toxicity Test". • Method C.1 ofCommissionRegulation (EC) No. 440/2008. StudyDirector: TestFacility:

Sponsor:

Harlan StudyNumber: Study Completion Date: SParr HarlanLaboratoriesLtd. ShardlowBusiness Park Shardlow Derbyshire DE722GD UK

41206435 26 April2013

Page 1of38Harlan StudyNumber: 41206435 Report Page2


## CONTENTS CONTENTS
....................................................................................................................................2 STUDY DIRECTOR STATEMENT OF GLP COMPLIANCE
....................................................3 QUALITYASSURANCE STATEMENT
......................................................................................4 SIGNATURE OF CONTRIBUTING SCIENTISTS
......................................................................5 SUMMARY
....................................................................................................................................6 Introduction
............................................................................................................................6 Methods
.................................................................................................................................6 Results
....................................................................................................................................6 GENERAL INFORMATION
.........................................................................................................7 Schedule
.................................................................................................................................7 Animal Welfare
......................................................................................................................7 Archiving
...............................................................................................................................7 1 INTRODUCTION AND PURPOSE
.....................................................................................8 1.1 Guidelines I

### Regulations
.......................................................................................... 8 2 TEST ITEM
........................................................................................................................... 8 2.1 TestItem
.................................................................................................................. 8 3 MATERIALS AND METHODS
...........................................................................................8 3.1 Definitions and Abbreviations
.................................................................................8 3.2 Test Species
.............................................................................................................9 3.3 · TestWater
................................................................................................................9 3.4 Procedure
............................................................................................................... 10 3.4.1 Range-finding Test
.................................................................................. 10 3.4.2 Definitive Test
......................................................................................... 10 4 RESULTS
............................................................................................................................ 13 4.1 Range-finding Test
................................................................................................ 13 4.2 Definitive Test
....................................................................................................... 13 4.2.1 VerificationofTest Concentrations
........................................................ 13 4.2.2 Mortality Data
......................................................................................... 13 4.2.3 Sub-Lethal Effects
................................................................................... 14 4.2.4 Observations onTest Item Solubility
...................................................... 14 4.2.5 Physico-Chemical Measurements
........................................................... 14 5 CONCLUSION
.................................................................................................................... 15 6 REFERENCES
.................................................................................................................... 16 TABLES
........................................................................................................................................ 17 Table 1 Cumulative Mortality Datainthe Range-finding Test
.......................................... 17 Table 2 CumulativeMortality Datainthe Definitive Test..
............................................... 18 FIGURES
...................................................................................................................................... 19 Figure 1 Concentration-Mortality Curve
............................................................................. 19 Figure 2 Toxicity Curve
.......................................................................................................20 APPENDICES
...............................................................................................................................21 Appendix 1 Appendix 2 Appendix 3 Appendix4 Typical Water Quality Characteristics
..................................................................21 Analytical Investigations
.......................................................................................23 Physico-Chemical Measurements
..........................................................................3 7 Monitoring Authority StatementofGLP Compliance
..........................................38Harlan StudyNumber: 41206435 Report


## STUDYDIRECTOR STATEMENT OF GLP COMPLIANCE Harlan Laboratories Ltd., ShardlowBusiness Park, Shardlow, Derbyshire, DE72 2GD, UK
Harlan StudyNumber: Study Title: 41206435 Acute Toxicity To RainbowTrout (Oncorhynchus mykiss) Page3

The study described was performed in compliance with UK GLP standards (Schedule 1, Good Laboratory Practice Regulations 1999 (SI 1999/3106 as amended by SI 2004/0994)). These Regulations are in accordance with GLP standards published as OECD Principles on Good Laboratory Practice (revised 1997, ENV/MC/CHEM(98)17). These principles are compatible with Good Laboratory Practice regulations specified by regulatory authorities throughout the European Community, the United States (EPA and FDA), and Japan (MHLW, MAFF and METI). This report fully and accurately reflects the procedures used and data generated. There were no circumstances consideredto have affected the integrity ofthe study orthe validity ofthe data. Study Director: SParrHarlan StudyNumber: 41206435 Report Page4 

## QUALITYASSURANCE STATEMENT Harlan Laboratories Ltd., ShardlowBusiness Park, Shardlow, Derbyshire, DE72 2GD, UK Harlan StudyNumber: StudyTitle: 41206435 Acute Toxicity To Rainbow Trout (Oncorhynchus mykiss)
The general facilities and activities are inspected at least once a year and the results are reported to the relevant responsible person and management. Study-relatedprocedures conducted atthe test facility were audited and inspected. The details of these audits and inspections are givenbelow. Reported to the Dates and Types ofQAInspections relevant·study Director and Test Facility Management Date ofInspection Type ofInspection Phase Inspected ReportDate 19 November 2012 StudyPlan N/A 19 November 2012 Verification 08 January 2013 Process

- based Test Item Preparation 08 January 2013 10 January 2013 Process

- based Test System 1 0 January 2013 Preparation 07 January 2013 Process

- based Exposure 07 January 2013 10 January 2013 Process

- based Assessment of 10 January 2013 Response 11 January 2013 Process

- based Chemical Analysis 11 January 2013 05 April2013 Report Audit N/A 05 April2013 This statement confirmsthat this report reflects the raw data and the procedures followed. QualityAssurance:



### .......**1:-::.f!..~l.** 

## 24 APR 2013Harlan StudyNumber: 41206435 Report Page 5


## SIGNATURE OFCONTRIBUTING SCIENTISTS
Analytical Chemistry: DMMullee Harlan Laboratories Ltd., ShardlowBusiness Park, Shardlow, Derbyshire, DE72 2GD, UK 

# /J

# 'IIf1 

## 20 

## l013

......

## (/...~.............................. Date:Harlan StudyNumber: 41206435 Report Page6


## SUMMARY Introduction A study was performed to assess the acute toxicity of the test item to rainbow trout (Oncorhynchus mykiss). 

### The method followed was designed to be compatible with the OECD Guidelines for Testing ofChemicals (1992) No 203, "Fish, Acute Toxicity Test" referenced as Method C.1 ofCommissionRegulation (EC) No. 440/2008. 

### Methods Following a preliminary range-finding test, fish were exposed, in groups ofseven, to an aqueous solution ofthe test item over a range ofconcentrations of 1.0, 1.8, 3.2, 5.6 and 10 mg/L for a period of 96 hours at a temperature of 13 °C to 14 °C under semi-static test conditions. The number ofmortalities and any sub-lethal effects ofexposure in each test and control vessel were determined 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours. 

### Results Analysis ofthe test preparations at 0 and 72 hours (fresh media) and at 24 and 96 hours (old media) showedthe following meanmeasuredtest concentrations: Nominal Concentration Measured Concentrations MeanMeasured Concentration (mg/L) (mg/L) (mg/L) 1.8 1.7-2.4 2.1 3.2 2.9-4.1 3.5 5.6 5.3-6.8 6.1 10 9.3

- 12 11 

### The 96-Hour LCso based on mean measured test concentrations was 6.5 mg/L with 95% confidence limits of4.6

- 10 mg!L. The Lowest Observed Effect Concentration was considered to be 3.5 mg!L. TheNo Observed Effect Concentrationwas 2.1 mg/L.Harlan StudyNumber: 41206435 Report


## GENERALINFORMATION Schedule Experimental Starting Date: Experimental CompletionDate: 

### Animal Welfare 11 February 2013 08 March 2013 Page 7

The study was designed and conducted to cause the minimum suffering or distress to the animals consistent with the scientific objectives and in accordance with the Harlan Laboratories Ltd, Shardlow, UKpolicy on animal welfare and the requirements ofthe United Kingdom's Animals (Scientific Procedures) Act 1986. The conduct ofthe study may be retrospectively reviewed, as part ofthe Harlan Laboratories Ltd, Shardlow, UK Ethical ReviewProcess. The study was conducted in accordance with the UK Home Office Guidance document on Regulatory Toxicology and Safety Evaluation Studies and the OECD guidance document on recognition, assessment and use ofclinical signs as humane endpoints for experimental animals used in safety evaluation. 

### Archiving Unless instructed otherwise by the Sponsor, the study plan (general studyplan and study specific supplement), all raw data (paper and electronic) and the final report will be retained iJ1 the Harlan Laboratories Ltd, Shardlow, UK archives for five years after which instructions will be sought as to further retention or disposal. Further retention or return ofthe data will be chargeable to the Sponsor. No data will be discarded without contacting the Sponsorto obtaintheirwritten consent.Harlan StudyNumber: 41206435 Report Page 8


## 1 INTRODUCTIONAND PURPOSE This report contains a description ofthe methods used and results obtained during a study to investigate the acute toxicity ofthe testitemto rainbowtrout (Oncorhynchus mykiss). Rainbow trout is a freshwater fish representative ofa wide variety ofnatural habitats, and can therefore be considered as animportantnon-target organisminfreshwater ecosystems. 

### 1.1 Guidelines I

### Regulations This study was designed to be compatible with the procedures indicated by the following internationally accepted guidelines andrecommendations:
• OECD Guidelines for Testing ofChemicals (1992) No 203 "Fish, Acute ToxicityTest". • Method C.1 ofCommissionRegulation (EC) No. 440/2008. 

## 2 TESTITEM 2.1 TestItem The integrity ofthe supplied data relating to the identity, purity and stability ofthe test item is theresponsibility ofthe Sponsor. Identification: Description: Batch: Purity: Expiry Date: Storage Conditions: 

## 3 MATERIALSAND METHODS 3.1 Definitions and Abbreviations LC50: The calculated concentration oftest item which results in 50% mortality withinthe exposure period. LOEC (Lowest Observed Effect Concentration): The lowesttestconcentrationatwhich a significanttoxic effect onthe test organisms is observedwithinthe exposureperiod.Harlan StudyNumber: 41206435

NOEC (No Observed .Effect Concentration):
Treatment:



### 3.2 Test Species Report Page 9

The highest test concentration at which no significant toxic effect on the test organisms is observed within the exposure period. Comprises test item treatments, control (test water only) and solvent control (ifapplicable).

The test was carried out using juvenile rainbow trout (Oncorhynchus mykiss). Fish were obtained from Brow Well Fisheries Limited, Hebden, near Skipton, Yorkshire, UK and maintained in-house since 4 February 2013. Fish were maintained in a glass fiber tank with a "single pass" water renewal system. Fish were acclimatized to test conditions from 20 February 2013 to 4 March 2013. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusktransitionperiods. The watertemperature was controlled at approximately 14

## oc with a dissolved oxygen content of greater than or equal to 10.5 mg 02/L. These parameters were recorded daily. The stock fish were fed commercial trout pellets which was discontinued approximately 24 hours prior to the start ofthe definitive test. There was 1mortality inthe 7 days priorto the start ofthe test and the fish had a mean standard length of4.6 em (sd 

### =0.2) and a mean weight of 1.17 g (sd 

### = 0.21) at the end ofthe definitive test. Based on the mean weight value this gave a loading rate of0.41 g bodyweight/liter (static volume). The diet and diluent water are considered not to contain any contaminant that would affect the integrity and outcome ofthe study. 

### 3.3 Test Water The test water used for both the range-finding and definitive tests was the same as that used to maintainthe stock fish. Laboratory tap water was dechlorinated by passage through an activated carbon filter (Purite Series 500) and partly softened (ElgaNimbus 1248D Duplex Water Softener) giving water with a total hardness ofapproximately 140 mg/L as CaC03. After dechlorination and softening the water was passed through a series ofcomputer controlled plate heat exchangers to achieve the required temperature. Typical water quality characteristics for the tap water as supplied, prior to dechlorination and softening, are given in Appendix 1.Harlan StudyNumber: 41206435 Report Page 10



### 3.4 Procedure 3.4.1 Range-finding Test The test concentrations to be used in the definitive test were determined by a preliminary rangefinding test. In the range-finding test fish were exposed to a series of nominal test concentrations of 1.0, 10 and 100 mg/L. An amount of test item (4000 mg) was dissolved in dechlorinated tap water and the volume adjusted to 1 liter to give a 4000 mg/L stock solution. Aliquots (5.0, 50 and 500 mL) ofthe 4000 mg/L stock solution were then each separately added to a final volume of 20 liters of dechlorinatedtap water to give the 1.0, 10 and 100 mg/L test concentrations respectively. The stock solution was inverted several times, and the prepared concentrations were stirred using a flat-bladed mixerfor approximately 1minute to ensure adequate mixing and homogeneity. In the range-finding test 3 fish were added to each 20 liter test and control vessel and maintained at 13 °C to 14 °C in a temperature controlled room with a photoperiod of 16 hours light and 8hours darkness with 20 minute dawn and dusktransition periods for a period of96 hours under static test conditions. The control group was maintained under identical conditions butnot exposed to the test item. Datafrom the control group was shared with similar concurrent studies. Each vessel was covered to reduce evaporation. After 3, 6, 24, 48, 72 and 96 hours any mortalities or sub-lethal effects ofexposure were determined by visual inspection ofthe test fish. A sample ofeachtest concentration was taken for chemical analysis at 0 and 24 hours in order to determine the stability of the test item under test conditions. All samples were stored at approximately -20 °C prior to analysis. Only concentrations within the range to be used for the definitive test were analyzed.
3.4.2 Definitive Test Based on the results ofthe range-finding test the following test concentrations were assigned to the definitive test: 1.0, 1.8, 3.2, 5.6 and 10 mg/L.
3.4.2.1 Experimental Preparation An amount oftest item (1000 mg) was dissolved in dechlorinated tap water and the volume adjusted to 1 liter to give a 1000 mg/L stock solution. Aliquots (20, 36, 64, 112 and 200 mL) ofHarlan StudyNumber: 41206435 Report Page 11

the 1000 mg/L stock solution were then each separately added to final volumes of20 liters of dechlorinated tap water to give the 1.0, 1.8, 3.2, 5.6 and 10 mg/L test concentrations respectively. The stock solution was inverted several times, and the prepared concentrations were stirred using a flat-bladed mixer for approximately 1minute to ensure adequate mixing and homogeneity. The concentration and stability ofthe test item in the testpreparations were verified by chemical analysis at 0 and 72 hours (fresh media) and at 24 and 96 hours (old media) hours (see Appendix 2).
3.4.2.2 Exposure Conditions As in the range-finding test, 20 liter glass exposure vessels were used for eachtest concentration. At the start ofthe test 7 fish were placed in each test vessel at random, in the test preparations. The test vessels were then covered to reduce evaporation and maintained at 13 

## oc to 14 °C in a temperature controlled room with a photoperiod of 16 hours light and 8 hours darkness with 20 minute dawn and dusk transition periods for a period of 96 hours. The test vessels were aerated via narrow bore glass tubes. The fish were not individually identified and received no food during exposure. The control group was maintained under identical conditions but not exposedto the test item. Datafrom the control group was shared with similar concurrent studies. A semi-static test regime was employed in the test involving a daily renewal of the test preparations to ensure that the concentrations of the test item remained near nominal and to preventthe build-up ofnitrogenous waste products. Any mortalities and sub-lethal effects ofexposure were recorded at 3, 6, 24, 48, 72 and 96 hours after the start ofexposure. The criteria ofdeath were taken to be the absence ofboth respiratory movement and response to physical stimulation.
3.4.2.3 Physico-Chemical Measurements The water temperature, pH and dissolved oxygen concentrations were recorded daily throughout the test. The measurements at 0 hours, and after each test mediarenewal at 24, 48 and 72 hours, represent those ofthe freshly prepared test preparations while the measurements taken prior to each test mediarenewal, and on termination ofthe·test after 96 hours, represent those ofthe used or 24-Hour old testpreparations. The pH and oxygen were measured using a Hach HQ30d Flexi handheld meter and the temperature was measured using a Hanna Instruments HI 93510 digital thermometer.Harlan StudyNumber: 41206435 Report Page 12

3.4.2.4 Verification ofTest Concentrations Water samples were taken from the control and all surviving test groups at 0 and 72 hours (fresh media) and at 24 and 96 hours (old media) for quantitative analysis. The 0, 24 and 72 hour samples were stored at approximately -20 °C prior to analysis withthe 96 hour samples. Duplicate samples and samples at 24 (fresh media), 48 (old and fresh media) and 72 hours (old media) were taken and stored at approximately -20 °C for further analysis ifnecessary. The method ofanalysis, recovery and testpreparation analyses are described in Appendix 2.
3.4.2.5 Evaluation ofData The LC50 values and associated confidence limits at 24 and 48 hours were calculated by the maximum-likelihood probit method (Finney, 1971) using the ToxCalc computer software package (ToxCalc, 1999). Probit analysis is usedwhere two ormore partial responses to exposure are shown.Harlan StudyNumber: 41206435 Report Page 13


## 4 RESULTS 4.1 Range-finding Test Cumulative mortality data from the exposure of rainbow trout to the test item during the range-finding test are given in Table 1. There were no sub-lethal effects ofexposure during the range-finding test. The results showed no mortalities at the test concentration of 1.0 mg/L. However, mortalities were observed at 10 and 100 mg/L. Based on this information test concentrations of 1.0, 1.8, 3.2, 5.6 and 10 mg/L were selected for the definitive test. Chemical analysis ofthe test preparations at 0 and 24 hours (see Appendix 2) showed measured concentrations of 99% to 106% ofnominal indicating that the test item was stable under test conditions. 

### 4.2 Definitive Test
4.2.1 Verification ofTest Concentrations Analysis ofthe test preparations at 0, 24, 72 and 96 hours (see Appendix 2) showed the test item to be stable between mediarenewal time points (i.e., 0to 24 hours and 72 to 96 hours). However, variation was observed when the 0 hour fresh media to the 72 hour fresh media were compared, and when the 24 hour old media to the 96 hour old media were compared. It was therefore considered justifiable to calculate the LC5o values in terms of the mean-measured test concentrations: Nominal Concentration Measured Concentrations MeanMeasured Concentration (mg/L) (mg/L) (mg/L) 1.8 1.7-2.4 2.1 3.2 2.9-4.1 3.5 5.6 5.3-6.8 6.1 10 9.3- 12 11


### 4.2.2 Mortality Data Cumulative mortality data from the exposure of rainbow trout to the test item during the definitive test are given in Table 2 and the relationship between percentage mortality and concentration at 96 hours is given inFigure 1.Harlan StudyNumber: 41206435 Report Page 14

Time (h) LCso (mg/L) 95% Confidence limits (mg/L) 3 >11


## - 6 

### >11


## - 24 >11 

## - 48 7.0 5.0- 12 72 6.5 4.6- 10 96 6.5 4.6- 10 The results ofthe definitive test showed the highest test concentration resulting in 0% mortality to be 2.1 mg/L and the lowest test concentration resulting in 100% to be greater than 11 mg/L. The Lowest Observed Effect Concentration (LOEC) was considered to be 3.5 mg/L. The No Observed Effect Concentration (NOEC) was 2.1 mg/L. The relationship between the median lethal concentration (LC50) and time 1s presented graphically in Figure 2.


### 4.2.3 Sub-Lethal Effects There were no sub-lethal effects ofexposure observed inthe test.


### 4.2.4 Observations on Test Item Solubility The testpreparations were observedto be clear colorless solutions for the duration ofthe test.


### 4.2.5 Physico-ChemicalMeasurements The results ofthe physico-chemical measurements are given in Appendix 3. Temperature was maintained at 13 

## oc to 14 °C throughout the test, while there were no treatment related differences for oxygen concentration orpH. The oxygen concentration in some of the test vessels was observed to have an air saturation value (ASV) in excess of 100%. This was considered to be due to the presence ofmicroscopic air bubbles in the media super-saturating the diluent and was considered not to have had an impact on the outcome or integrity ofthe test as no adverse effects were observed in the control group.Harlan StudyNumber: 41206435 Report Page 15


## 5 CONCLUSION Based on the mean measured test concentrations ofthe test media the acute toxicity ofthe test item to rainbow trout gave a 96-Hour LC5

### ovalue of6.5 mg/L with 95% confidence limits of 4.6- 10 mg/L. The Lowest Observed Effect Concentration was considered to be 3.5 mg/L. The No Observed Effect Concentrationwas 2.1 mg/L.Harlan StudyNumber: 41206435 Report Page 16 

# 6 REFERENCES FINNEY, D.J. (1978) Statistical Methods 

### in Biological Assay. 3rd ed. London: Charles Griffin. TOXCALC VERSION 5.0.23C (1999), Tidepool Scientific Software, McKinleyville, CA 95519, USA.Harlan StudyNumber: 41206435 Report Page 17


## TABLES Table 1 CumulativeMortalityDatain the Range-findingTest
Nominal Cumulative Mortality Concentration (Initial Population

### =3) (mg/L) 3 Hours 6 Hours 24 Hours 48 Hours 72 Hours 96 Hours Control 0 0 0 0 0 0 1.0 0 0 0 0 0 0 10 0 0 1 3 3 3 100 0 0 3 3 3 3Harlan StudyNumber: 41206435 Report Page 18



### Table2 CumulativeMortality Datain theDefinitive Test Nominal Mean Cumulative Mortality (Initial Population = 7) 0 /o Concentration Measured Mortality (mg/L) Concentration 3 6 24 48 72 96 96 (mg/L) Hours Hours Hours Hours Hours Hours Hours Control 

### - 0 0 0 0 0 0 0 1.0 • 0 0 0 0 0 0 0 

### - 1.8 2.1 0 0 0 0 0 0 0 3.2 3.5 0 0 0 1 1 1 14 5.6 6.1 0 0 0 2 3 3 43 10 11 0 0 1 6 6 6 86

•No Mean Measured concentration determined as only the No ObservedEffect Concentration (1.8 mg/L) and above were analyzed.Harlan StudyNumber: 41206435 Report Page 19


## FIGURES Figure 1 Concentration-Mortality Curve

99.9

99

90

70 50 30

10

0.1
0.01**~----~~--------------~------------------------~** 1 10 100 Concentration (mg!L)Harlan StudyNumber: 41206435 Report Page 20
Figure 2 Toxicity Curve

10

10 100 Lc50 Value (mg!L)Harlan StudyNumber: 41206435 Report Page 21


## APPENDICES Appendix 1 Typical Water Quality Characteristics
WATER SUPPLY ZONE ZDB13 ALVASTON REPORTING PERIOD: 01/01/2010-31/12/2010 NO. CONCENTRATION ORVALUE SAMPLES PARAMETER ANALYSIS SAMPLES (ALL SAMPLES) PCV+ CONTRAVENING PCV UNITS TAKEN MINIMUM AVERAGE MAXIMUM NUMBER PERCENT 1,2 Dichloroethane**~g/1** 40 <0.080 <0.080 <0.080 3 0 0.00 Aluminium**~g/1** 38 <5.000 <6.684 12.000 200 0 0.00 Ametryn**~g/1** 8 <0.003 <0.003 <0.003 0.1 0 0.00 Antimony**~g/1** 8 0.180 0.531 1.200 5 0 0.00 Arsenic**~g/1** 8 <0.370 <0.370 <0.370 10 0 0.00 Atrazine**~g/1** 8 <0.003 <0.003 <0.003 0.1 0 0.00 Azinphos methyl**~g/1** 8 <0.004 <0.004 <0.004 0.1 0 0.00 Benzene**~g/1** 40 <0.060 <0.060 <0.060 1 0 0.00 Benzo 3,4-Pyrene**~g/1** 8 <0.001 <0.001 <0.001 0.01 0 0.00 Boron mg/1 40 <0.017 0.020 0.038 1 0 0.00 Bromacil**~g/1** 8 <0.004 <0.004 <0.004 0.1 0 0.00 Bromate**~g/1** 8 <0.600 <0.600 <0.600 10 0 0.00 Cadmium**~g/1** 8 0.080 0.121 0.160 5 0 0.00 Carbenazim**~g/1** 8 <0.002 <0.002 <0.002 0.1 0 0.00 Carbetamide**~g/1** 8 <0.006 <0.006 <0.006 0.1 0 0.00 Carbophenophion**~g/1** 8 <0.012 <0.012 <0.012 0.1 0 0.00 Chlorfenvinphos**~g/1** 8 <0.005 <0.005 <0.005 0.1 0 0.00 Chlorotoluron**~g/1** 8 <0.004 <0.004 <0.004 0.1 0 0.00 Chlorpyriphos Ethyl**~g/1** 8 <0.010 <0.010 <0.010 0.1 0 0.00 Chlorpyriphos Methyl**~g/1** 8 <0.015 <0.015 <0.015 0.1 0 0.00 Chromium**~g/1** 8 <0.700 <0.700 0.700 50 0 0.00 Colour mg/1 Pt/Co 38 <0.400 <0.608 2.600 20 0 0.00 Copper mg/1 8 <0.003 <0.032 0.130 2 0 0.00 Cyanazine**~g/1** 8 <0.005 <0.005 <0.005 0.1 0 0.00 Cyanide**~g/1** 40 <0.300 0.740 1.600 50 0 0.00 Demeton-S-Methyl**~g/1** 8 <0.006 <0.006 <0.006 0.1 0 0.00 Diazinon**~g/1** 8 <0.006 <0.006 <0.006 0.1 0 0.00 DichloNos**~g/1** 8 <0.008 <0.008 <0.008 0.1 0 0.00 Diflufenican**~g/1** 8 <0.005 <0.005 <0.005 0.1 0 0.00 Dimethoate**~g/1** 8 <0.005 <0.005 <0.005 0.1 0 0.00 Diuron**~g/1** 8 <0.003 <0.003 <0.003 0.1 0 0.00 E. coli No./100 ml 204 0.000 0.000 0.000 0 0 0.00 Enterococci No./100 ml 8 0.000 0.000 0.000 0 0 0.00 Ethofumesate**~g/1** 8 <0.006 <0.006 <0.006 0.1 0 0.00 Fenpropidin**~g/1** 8 <0.004 <0.004 <0.004 0.1 0 0.00 Fenpropimorph**~g/1** 8 <0.002 <0.002 <0.002 0.1 0 0.00 Fluoride mg/1 8 0.270 0.331 0.414 1.5 0 0.00 Flutriafol**~g/1** 8 <0.007 <0.007 <0.007 0.1 0 0.00 Heptenophos**~g/1** 8 <0.005 <0.005 <0.005 0.1 0 0.00 Iron**~g/1** 76 <10.000 <26.579 488.000 200 1 1.32 lsoproturon**~g/1** 8 <0.002 <0.002 <0.002 0.1 0 0.00 Lead**~g/1** 8 <0.500 <2.288 10.000 25 0 0.00 Linuron i-iQ/1 8 <0.005 <0.005 <0.005 0.1 0 0.00 Malathion**~g/1** 8 <0.004 <0.004 <0.004 0.1 0 0.00 Manganese**~g/1** 38 <1.800 <1.987 5.600 50 0 0.00 Mercury**~g/1** 40 <0.012 0.018 0.199 1 0 0.00 Metaldehyde**~g/1** 32 0.004 0.009 0.027 0.1 0 0.00 Metamitron**~g/1** 8 <0.003 <0.003 <0.003 0.1 0 0.00 Metazachlor**~g/1** 8 <0.003 <0.003 <0.003 0.1 0 0.00 Methabenzthiazuron**~g/1** 8 <0.003 <0.003 <0.003 0.1 0 0.00 Mevinphos**~g/1** 8 <0.004 <0.004 <0.004 0.1 0 0.00 Monolinuron**~g/1** 8 <0.004 <0.004 <0.004 0.1 0 0.00 +PCV Prescribed concentration or value for mandatory parameters and specified limit or value for non-mandatory parameters (indicators and disinfectants).Harlan StudyNumber: 41206435 Report Page 22 Typical Water Quality Characteristics

WATER SUPPLY ZONE ZDB13 ALVASTON REPORTING PERIOD: 01/01/2010-31/12/2010 NO. CONCENTRATION OR VALUE SAMPLES PARAMETER ANALYSIS SAMPLES (ALL SAMPLES) PCV+ CONTRAVENING PCV UNITS TAKEN MINIMUM AVERAGE MAXIMUM NUMBER PERCENT Nickel IJg/1 8 2.300 3.288 5.800 20 0 0.00 Nitrate mg/1 8 9.940 13.105 16.700 50 0 0.00 Nitrite mg/1 8 <0.003 <0.005 <0.009 0.5 0 0.00 Nitrite/Nitrate Calculated -1 8 0.200 0.276 0.370 <1' 0 0.00 Odour Dilution 42 0.000 0.024 1.000 -2 1 2.38 Number PAH IJg/1 8 0.000 0.001 0.002 0.1 0 0.00 Parathion IJg/1 8 <0.006 <0.006 <0.006 0.1 0 0.00 Pendimethalin IJg/1 8 <0.005 <0.005 <0.005 0.1 0 0.00 pH pH value 76 7.020 7.497 7.870 Max 9.5, 0 0.00 Min 6.5 Phorate IJg/1 8 <0.009 <0.009 <0.009 0.1 0 0.00 Phosalone IJg/1 8 <0.007 <0.007 <0.007 0.1 0 0.00 Pirimicarb IJg/1 8 <0.003 <0.003 <0.003 0.1 0 0.00 Pirimiphos-Methyl IJg/1 8 <0.009 <0.009 <0.009 0.1 0 0.00 Prometryn IJg/1 8 <0.003 <0.003 <0.003 0.1 0 0.00 Propachlor IJg/1 8 <0.005 <0.005 <0.005 0.1 0 0.00 Propazine IJg/1 8 <0.004 <0.004 <0.004 0.1 0 0.00 Propetamphos IJg/1 8 <0.007 <0.007 <0.007 0.1 0 0.00 Propiconazole IJg/1 8 <0.010 <0.010 <0.010 0.1 0 0.00 Propyzamide IJg/1 8 <0.004 <0.004 <0.004 0.1 0 0.00 Selenium IJg/1 8 <0.220 <0.546 0.740 10 0 0.00 Simazine IJg/1 8 <0.007 <0.007 <0.007 0.1 0 0.00 Sodium mg/1 8 14.000 25.250 30.000 200 0 0.00 Sulphate mg/1 40 23.800 50.345 99.800 25 0 0.00 Taste Dilution 42 0.000 0.000 0.000 2 0 0.00 Number 

### - Terbutryn IJg/1 8 <0.003 <0.003 <0.003 0.1 0 0.00 Tetrachloroethene and IJg/1 8 0.000 0.000 0.000 10 0 0.00 Trichloroethene Tetrachloromethane IJg/1 8 <0.018 <0.021 <0.030 3 0 0.00 Total Pesticides IJg/1 32 0.000 0.009 0.027 0.5 0 0.00 Total Trihalomethanes IJg/1 8 19.440 25.449 31.580 100 0 0.00 Triadimefon IJg/1 8 <0.009 <0.009 <0.009 0.1 0 0.00 Triallate IJg/1 7 <0.008 <0.014 <0.050 0.1 0 0.00 Triazophos IJg/1 8 <0.003 <0.003 <0.003 0.1 0 0.00 Trietazine IJg/1 8 <0.006 <0.006 <0.006 0.1 0 0.00 Turbidity NTU 76 <0.030 <0.135 3.400 4 0 0.00 Free Chlorine mg/1 204 0.010 0.207 0.630 

### - 3


### - - Total Chlorine mg/1 204 0.040 0.270 0.710 

### - 3


### - - Ammonium mg/1 38 <0.009 <0.012 0.049 0.5 0 0.00 Chloride mg/1 40 9.660 21.052 50.800 250 0 0.00 Clostridium perfringens No./100 ml 38 0.000 0.000 0.000 0 0 0.00 Coliform Bacteria No./100 ml 204 0.000 0.000 0.000 0 0 0.00 Colony Count after 48 No./100 ml 76 0.000 0.789 31.000 

### - 4


### - - hours at 37°C Colony Count after 72 No. /100 ml 76 0.000 2.039 85.000 

### - 4


### - - hours at22°C Conductivity iJS/cm at 20°C 38 315.000 419.000 486.000 2500 0 0.00 Gross Alpha Activity Bq/1 8 0.032 0.069 0.090 0.1 5 Gross Beta Activity Bq/1 8 <0.001 <0.036 0.053 15 Total Organic Carbon mg/1 40 <0.130 0.920 2.020 

### - 4


### - - +PCV Prescribed concentration orvalue for mandatory parameters and specified limit orvalue for non-mandatory parameters (indicators and disinfectants). 1 The formula forthe calculated Nitrate/Nitrite level is (Nitrate]/50 + [Nitrite]/3. This number has no units. 2 Acceptable to customers and no abnormal change. 3 No numerical limit applies. 4 No abnormal change. 5 Guidance levels only. Monitored forthe calculation ofTotal Indicative Dose (TID), which is the regulatory parameter. No samples exceeded the TID in 2010.Harlan Study Number: 41206435 Report Page 23
Appendix 2 Analytical Investigations 

## Analytical Investigations CONTENTS
CONTENTS
..................................................................................................................................23 1 MATERIALS AND METHODS
.........................................................................................24 1.1 Introduction
...........................................................................................................24 1.2 Test Item
................................................................................................................24 1.3 Analytical Standard
...............................................................................................24 1.4 Analytical Procedure
.............................................................................................24 1.4.1 Storage
.....................................................................................................24 1.4.2 Reagents and Solvents
.............................................................................24 1.4.3 Preparation ofCalibration Standards
.......................................................25 1.4.4 Preparation ofLinearity Standards
..........................................................25 1.4.5 Preparation ofSpiked Recovery Samples
...............................................25 1.4.6 Preparation ofTest Samples
....................................................................25 1.4.7 Instrumental Parameters
..........................................................................26 1.4.8 Calculations
.............................................................................................27 2 RESULTS
............................................................................................................................29 2.1 Range-Finding Test
...............................................................................................29 2.2 Definitive Test
.......................................................................................................29 2.2.1 Validation ofthe Analytical Method
.......................................................29 2.2.2 Test Samples
............................................................................................29 3 DISCUSSION
......................................................................................................................30 TABLES
........................................................................................................................................31 Table 1 Preparation ofTest Samples
.................................................................................31 Table 2 Calibration Standard Data
....................................................................................31 Table 3 Results for Range-Finding Samples
.....................................................................32 Table 4 Results for Spiked Recovery Samples
..................................................................32 Table 5 Results for Test Samples
......................................................................................33 FIGURES
......................................................................................................................................34 Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 CalibrationPlot
......................................................................................................34 Chromatogram ofCalibration Solution (1.5 mg/L)
...............................................35 Chromatogram ofBiological Control Sample
.......................................................35 Chromatogram ofTestSample (1.8 mg/L)
...........................................................36 Chromatogram ofTest Sample (10 mg/L)
............................................................36Harlan StudyNumber: 41206435 Report Page24 Analytical Investigations 

## 1 MATERIALSAND METHODS 1.1 Introduction The test item concentration in the test samples was determined by direct injection into an electrospray mass spectrometer (MS) using an external standard. The method was developed by the Department ofAnalytical Services, Harlan Laboratories Ltd, Shardlow, UK. 

### 1.2 TestItem Thetest itemis described inthe biologicalpart ofthis study. 

### 1.3 Analytical Standard The test item described in the biological part of this study was also used as the analytical standard. 

### 1.4 Analytical Procedure
1.4.1 Storage The test samples were stored at approximately -20 °C priorto analysis.


### 1.4.2 Reagents and Solvents Water: Fisher Scientific, HPLC-MS grade. Testmedium: as described inthe biologicalpart ofthis study. Solvents: methanol, Romil, UG grade. Reagents: formic acid, Fisher Scientific, analytical reagent grade.Harlan StudyNumber: 41206435 Report Page 25 Analytical Investigations 1.4.3 Preparation ofCalibration Standards 

### The test item (nominal 100 mg) was dissolved in (50:50 v/v) methanol:test medium (100 mL) to prepare a stock solution with a concentration of 1000 mg/L. This stock solution was further diluted with (50:50 v/v) methanol:test medium to produce a solution of 10 mg/L. Defined volumes of this solution were then diluted with (50:50 v/v) methanol:test medium to obtain calibration standards in the range of0.10 to 1.0 mg/L. A second series ofcalibration standards was similarly prepared in the range of0.30 to 1.5 mg/L. These solutions were used to determine the recovery and test sample concentrations.
1.4.4 Preparation ofLinearity Standards 

### The test item (nominal 100 mg) was dissolved in (50:50 v/v) methanol:test medium (100 mL) to prepare a stock solution with a concentration of 1000 mg/L. This stock solution was further diluted with (50:50 v/v) methanol:test medium to produce a solution of 10 mg/L. Defined volumes of this solution were then diluted with (50:50 v/v) methanol:test medium to obtain calibration standards in the range of0.10 to 1.0mg/L. A second series ofcalibration standards was similarly prepared in the range of0.30 to 1.5 mg/L. These standards were used to evaluate the linearity ofthe analytical system.
1.4.5 Preparation ofSpiked Recovery Samples 

### To demonstrate the validity ofthe analytical procedure, volumes oftest medium were spiked with the test item and its recovery was assessed. The test item (nominal 100 mg) was initially dissolved in (50:50 v/v) methanol:test medium to prepare a stock solution with a concentration of 1000 mg/L. This stock solution was further diluted with (50:50 v/v) methanol:test medium to produce a solution of 100 mg/L. A defined volume ofthis stock solution was diluted with test medium to obtain spiked recovery samples at a concentration of 1.0 mg/L. Five replicates were prepared and subjected to the same treatment as the test samples. In addition, test medium withoutthe addition ofthe test item (synthetic control) was also analyzed.
1.4.6 Preparation ofTest Samples 

### The test samples were thawed with the aid of sonication. They were initially diluted with methanol and then further diluted into the calibration range with (50:50 v/v) methanol:test medium as shown inTable 1.Harlan StudyNumber: 41206435 Report Analytical Investigations 1.4.7 Instrumental Parameters Autosampler: Pump: Detector: Column: Mobilephase: Flowrate: Injectionvolume: Ionization mode: Polarity: Capillary voltage: Gas temperature: Nebulizerpressure: Fragmentorvoltage: Drying gas flow: Data acquisition mode: SIM ion: Agilent 1200 Series Agilent 1200 Series Agilent 1200 Series MSD
no column water:methanol:formic acid (500:500:0.5, v/v/v) 0.5 mL!minute 10 J!L ESI positive 1000V 350

## oc 40 psi 100V 10 Llmin Single IonMonitoring (SIM) Page 26Harlan StudyNumber: 41206435 Report Page 27 Analytical Investigations

1.4.8 Calculations
A calibration curve was prepared using a quadratic function ofthe form shown in equation ( 1 ). The calibration data is presented in Table 2, and Figure 1. From the calibration curve, the concentration "x" of the test item in an injected sample was calculated using the following equation:

### -b+~b**2** -4·a·(c-y) X=---'--------'-- (1) 2·a where x: y: a, b, c: concentrationofthetest item ininjected sample [mg/L] average peakarea ofthe testitem in injected sample [counts] constants

The concentration of the test item in the test samples was calculated using the following equation:

c 

## = X 

### ·F R/100 where c: 

### x: F: R: (2) concentrationofthetest iteminthetest sample [mg/L] concentrationofthe testiteminthe injected sample solution [mg/L] samplepreparationfactor recoveryrate [%] (Ifnot correcting for recovery thenR is taken as 1 00)

The recovery rate (R) of the spiked recovery samples was calculated usmg the following equation: R =-c-·100 cfort where R: c: (3)
recoveryrate [%] determined concentrationofthe testitem inthe spikedrecovery sample [mg/L] fortified concentrationofthetestitem inthe spikedrecovery sample [mg/L]Harlan StudyNumber: 41206435 Report Page 28 Analytical Investigations

The percentage ofnominal concentration ofthe test item inthe test samples was calculatedusing the following equation: %nominal=_c_·100 cnom (4) where Cnom: nominal concentration inthe test sample [mg/L]Harlan StudyNumber: 41206435 Report Page 29 Analytical Investigations 

## 2 RESULTS 2.1 Range-Finding Test The analytical methodology was not validated priorto the analysis ofrange-finding samples; full validation was later conducted specifically over the concentration range to be used in the definitive test. The results for the range-finding test samples are summarized in Table 3. 

### 2.2 Definitive Test 2.2.1 Validation ofthe Analytical Method Linearity The calibration data for the calibration standards ofthe test item is given in Table 2 and Figure 1. The data was found to have a quadratic correlation within the calibration range of 0to 1.49 mg/L. The R2 fit ofthe calibration curve to the data was 0.9993, and was considered to be acceptable. 

### Accuracy (Recovery) and Precision A set of recovery samples accurately fortified at a relevant concentration of test item was prepared five-fold and analyzed. The results obtained for the concentration ofthe test item inthe recovery samples are presented in Table 4. The method was considered to be sufficiently accurate and precise for the purposes ofthis test. The test sample results were not corrected for recovery. 

### Limit ofQuantitation (LOQ) The limit ofquantitation (LOQ) was determined by calculating the concentration equivalent to ten times the standard deviation of five synthetic controls. Using this method the limit of quantitation was determined to be 0.28 mg/L.


### 2.2.2 Test Samples The results obtained for the concentration ofthe test item found in the test samples are presented in Table 5. Typical chromatograms are shown in Figures 2 to 5.Harlan StudyNumber: 41206435 Report Page 30 Analytical Investigations 

## 3 DISCUSSION The detection system was found to have acceptable correlation to the fit ofa quadratic equation. The analytical procedure was found to have acceptable recoveries oftest item in test medium. The method ofanalysis was validated andprovento be suitable for use.Harlan StudyNumber: 41206435 Report Page 31 Analytical Investigations 

## TABLES Table 1 Preparation ofTest Samples
Nominal Sample Sample FinalNominal Concentration of Volume FinalVolume Preparation Concentration ofTest TestItem Factor Item

[mg/L] [mL] [mL] F [mg/L] Control 5 10 2 

### - 1.8 5 10 2 0.90 3.2 5 10, then 4 0.80**5~10** 5.6 5 10, then 6 0.93**5~15** 10 5 10, then 10 1.0**4~20**



### Table2 Calibration StandardData
Concentration ofTestItem Area [mg/L] [counts] 0 4.773 X 104 0.0970 5.609 X 104 0.298 7.883 X 104 0.485 9.313 X 104 0.695 1.125 X 105 1.490 1.657 X 105Harlan StudyNumber: 41206435 Report Page 32 Analytical Investigations 

### Table3 Results forRange-Finding Samples Nominal Measured Sample Determined Concentration of Concentration Preparation ·Concentration TestItem in ofTestItem in Factor ofTest Item in %ofNominal TimePoint Range-Finding SampleVial Range-Finding Concentration Sample Sample
X F c Cnom (hours] (mg/L] [mg/L] [mg/L] [%] 0 1.0 0.497 2 0.994 99 10 1.06 10 10.6 106 24 1.0 0.501 2 1.02 102 10 1.05 10 10.5 105



### Table4 Results for SpikedRecovery Samples Nominal Fortified Measured Sample Determined Analytical Concentration Concentration Concentration Preparation Concentration Recovery Precision of ofTestItem in ofTestItem in Factor ofTestItem in Rate (Relative TestItem the Spiked the Sample the Spiked Standard Sample Vial Sample Deviation of Recovery) Cnom Cfort X F c R • [mg/L] [mg/L] [mg/L] [mg/L] [%] [%]

1.0 0.978 0.421 2 0.842 0.439 2 0.878 0.436 2 0.872 89 3.2 0.432 2 0.864 0.460 2 0.919 Acceptance Target 80-120 <10
Forreporting purposes, x has been calculated from cretrospectively.Harlan StudyNumber: 41206435 Report Analytical Investigations 

### Table5 Results forTest Samples
Nominal Measured Sample Concentration Concentration Preparation Time Point

[hours] 0 (Fresh Media)

24 (Old Media)

72 (Fresh Media)

96 (Old Media)

LOQ of ofTest Item in Test Item in SampleVial Test Sample
Cnom X [mg/L] [mg/L] Control <LOQ 1.8 0.935 3.2 0.763 5.6 0.882 10 0.933 Control <LOQ 1.8 0.855 3.2 0.723 5.6 0.890 10 0.943 Control <LOQ 1.8 1.21 3.2 1.01 5.6 1.14 10 1.19 Control <LOQ 1.8 1.15 3.2 1.00 5.6 1.14 10 1.19 Limit ofQuantitation not applicable Forreporting purposes, x has been calculatedfrom c retrospectively. Factor

F
2 2 4 6 10 2 2 4 6 10 2 2 4 6 10 2 2 4 6 10 Page 33

Determined Concentration ofTest Item in %ofNominal

Test Sample Concentration

c [mg/L] [%] <LOQ 

## -

1.87 104 3.05 95 5.29 94 9.33 93 <LOQ 

### -

1.71 95 2.89 90 5.34 95 9.43 94 <LOQ 

### -

2.42 134 4.05 127 6.82 122 11.9 119 <LOQ


### -

2.29 127 4.00 125 6.84 122 11.9 119Harlan StudyNumber: 41206435 Analytical Investigations 

## FIGURES Figure 1 Calibration Plot :§' 180000 ·c: 2- 160000 co**~** <( 140000 -"' co Q) 120000 c.. 100000 80000 60000 40000 20000 00.0 Report

y=-1.757E+04x2 + 1.057E+05x + 4.724E+04 R

2 =9.993E-01

0.5 1.0 1.5

Concentration (mg/L) Page 34Harlan StudyNumber: 41206435 Report Page 35 Analytical Investigations 

### Figure 2 Chromatogram ofCalibration Solution (1.5 mg/L)

0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 min 

### Figure 3 Chromatogram ofBiological Control Sample*

55000

50000

45000

«1000

35000

30000

0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 min

•Backgroundinterference level oftbe response. Nottest item.Harlan StudyNumber: 41206435 Report Page 36 Analytical Investigations 

### Figure 4 Chromatogram ofTest Sample (1.8 mg/L)

Figure 5 Chromatogram ofTest Sample (10 mg/L)

0.1 02 0.3 0.4 0.5 0.6 0.7 0.8 0.9 minHarlan StudyNumber: 41206435 Report Page 37


### Appendix 3 Physico-ChemicalMeasurements
Nominal Time (Hours) Concentration 0 Hours (Fresh Media) 24 Hours (Old Media) 24 Hours (Fresh Media) (mg/L) pH mg % T°C pH mg % T°C pH mg % T°C 02/L ASV* 0 2 /L ASV* 02/L ASV* Control 8.0 11.3 108 13 8.1 9.6 93 14 8.0 10.8 105 14 1.0 7.8 10.9 106 14 8.2 9.4 91 14 8.0 10.8 105 14 1.8 7.8 11.0 107 14 8.2 9.4 91 14 7.9 10.7 104 14 3.2 7.8 11.1 108 14 8.2 9.6 93 14 7.9 10.5 100 13 5.6 7.8 11.1 108 14 8.2- 9.3 90 14 7.9 10.5 102 14 10 7.8 11.1 108 14 8.2 9.3 90 14 7.9 10.5 102 14
Nominal Time (Hours) Concentration 48 Hours (Old Media) 48 Hours (Fresh Media) 72 Hours (Old Media) (mg/L) pH mg % T°C pH mg % T°C pH mg % T°C 02/L ASV* 02/L ASV* 02/L ASV* Control 8.1 9.7 94 14 8.0 10.7 104 14 8.2 9.9 96 14 1.0 8.2 9.4 91 14 8.0 10.6 103 14 8.2 9.4 91 14 1.8 8.2 9.3 90 14 8.0 10.5 102 14 8.2 9.5 92 14 3.2 8.3 9.4 91 14 8.0 10.5 102 14 8.2 9.4 91 14 5.6 8.2 9.4 91 14 7.9 10.4 101 14 8.3 9.2 89 14 10 8.2 9.2 89 14 7.9 10.4 101 14 8.2 9.0 87 14
Nominal Time (Hours) Concentration 72 Hours (Fresh Media) 96 Hours (Old Media) (mg/L) pH mg % T°C pH mg % T°C 0 2

/L ASV* 0 2 /L ASV* Control 8.0 10.7 104 14 8.2 9.9 96 14 1.0 8.0 10.6 103 14 8.2 9.5 92 14 1.8 8.0 10.5 102 14 8.2 9.5 92 14 3.2 8.0 10.4 101 14 8.3 9.4 91 14 5.6 7.9 10.4 101 14 8.3 9.2 89 14 10 8.0 10.3 98 13 8.3 9.1 88 14



### *

ASV = Dissolved oxygen concentration expressed as apercentage ofAir Saturation ValueHarlan StudyNumber: 41206435 Report


### Appendix 4 MonitoringAuthority Statement ofGLP Compliance

Department ofHealth
THE DEPARTMENT OF HEALTH OF THE GOVERNMENT OF THE UNITED KINGDOM GOOD LABORATORY PRACTICE

STATEMENT OF COMPLIANCE IN ACCORDANCE WITH DIRECTIVE 2004/9/EC TEST FACILITY Harlan Laboratories Ltd Shardlow Business Park London Road Shardlow Derby DE72 2GD

DATE OF INSPECTION

10 July 2012 TEST TYPE(S) Analytical/Clinical Chemistry Environmental Toxicity Environmental Fate Mutagenicity Phys/Chem. Tests Toxicology

An inspection for compliance with the Principles of Good Laboratory Practice was carried out at the above test facility as part of the UK Good Laboratory Practice Compliance Monitoring Programme. This statement confirms that, on the date of issue, the UK Good Laboratory Practice Monitoring Authority were satisfied that the above test facility was operating in compliance with the OECD Principles of Good Laboratory Practice. This statement constitutes a Good Laboratory Practice Instrument (as defined in the UK Good Laboratory Practice Regulations 1999).

Dr. Andrew J. Gray Head, UK GLP Monitoring Authority Page 38