Page 1 of 54 

TITLE/COVER PAGE 

PHYSICOCHEMICAL PROPERTIES 

Sponsor Research Laboratory 

Huntingdon Life Sciences, Ltd. 

Woolley Road 

Alconbury 

Huntingdon 

Cambridgeshire 

PE28 4HS 

ENGLAND 

Final: 23 April 2007 
: 2 : 

TABLE OF CONTENTS 

Page 

TITLE/COVER PAGE........................................................................................................ 1 

TABLE OF CONTENTS.................................................................................................... 2 

COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS ................. 3 

QUALITY ASSURANCE STATEMENT ......................................................................... 4 

CONTRIBUTING SCIENTISTS........................................................................................ 5 

SUMMARY ........................................................................................................................ 6 

INTRODUCTION............................................................................................................... 7 

TEST SUBSTANCE........................................................................................................... 8 

PHYSICOCHEMICAL PROPERTIES 

Vapour pressure ............................................................................................................... 9 

Surface tension................................................................................................................. 15 

Relative self-ignition temperature for solids.................................................................... 17 

Particle size distribution................................................................................................... 19 

Soil adsorption ................................................................................................................. 21 

DEVIATIONS FROM PROTOCOL .................................................................................. 28 

MAINTENANCE OF RECORDS...................................................................................... 28 

REFERENCES.................................................................................................................... 29 

APPENDIX 

1. Eye Research Centre GLP Compliance Statement 2005.............................................. 30 

2. Certificate of Analysis.................................................................................................. 32 

3. Study Protocol .............................................................................................................. 34 


: 5 : 

CONTRIBUTING SCIENTISTS 

Physicochemical Properties 

The following staff member has reviewed this report. 

A. L. Comb, B.Sc., Ph.D. 

(Scientific Manager, Product Chemistry) 

The following staff were responsible for the conduct of the work and reporting of the results. 

P. Sydney, B.Sc., M.Sc. 

(Study Director, Product Chemistry) 

C. Pointer 

(Scientist, Product Chemistry) 

C. Steil, B.Sc. 

(Scientist, Product Chemistry) 

P. Woods, H.N.C. 

(Scientist, Product Chemistry) 
: 6 : 

SUMMARY 

s performed to determine certain specified physicochemical properties of 

 The study was performed in accordance with the tests described in the EEC 

the Determination of Physicochemical Properties (Directive 92/69/EEC) and the 

OECD Guidelines for the Testing of Chemicals. 

The physicochemical properties which have been determined in this study are detailed below, 

together with the respective guideline methods followed and the result obtained for each test. 

EEC OECD Test Result 

Method 

no. 

Guideline 

no. 

 

A.4 104 Vapour pressure 4 x 10-9 Pa at 25°C

A.5 115 Surface tension 66.5 mN/m (90% saturated aqueous 

solution) 

 

A.16 - Relative self-ignition 

temperature for solids 

None below the melting 

temperature (approximately 150°C) 

 

- - Particle size distribution 0% w/w is smaller than 10 μm 

 

C.19 121 Soil adsorption LogKoc = 3.7 (immobile in soil) 
: 7 : 

INTRODUCTION 

was designed to determine certain specified physicochemical properties of 

 Information on physicochemical properties is important in the assessment of 

effects of a substance both at work and in the environment. 

The study was conducted in compliance with EEC Methods for the Determination of 

Physicochemical Properties (Directive 92/69/EEC) and the OECD Guidelines for the Testing 

of Chemicals. The physicochemical properties determined were vapour pressure, surface 

tension, relative self-ignition temperature for solids, particle size distribution and soil 

adsorption. 

The protocol was approved by the Study Monitor for the Sponsor on 8 September 2006, and 

by the Sponsor, the Study Director and Huntingdon Life Sciences Management on 

11 September 2006. 

The experimental start and completion dates were 14 September 2006 and 11 October 2006, 

respectively. 

Location of study : Huntingdon Life Sciences, Ltd. 

 Eye Research Centre 

 Eye 

 Suffolk 

 IP23 7PX 
: 8 : 

TEST SUBSTANCE 

Identity: 

Chemical name: 

Molecular formula: 

Molecular weight: 

CAS number: 

Lot number: 

Expiry date: 14 May 2007 

Purity: 

Appearance: 

Storage conditions: Ambient temperature in a dark environmentally 

controlled area 

Date of sample receipt: 31 July 2006 

Source: 

Structure: 
: 9 : 

PHYSICOCHEMICAL PROPERTIES 

VAPOUR PRESSURE 

(EEC Method A.4 and OECD Guideline 104)

METHOD 

The vapour pressure was determined using a vapour pressure balance. 

DEFINITION AND UNITS 

The vapour pressure of a substance is defined as the saturation pressure above a solid or 

liquid substance. At the thermodynamic equilibrium, the vapour pressure of a pure substance 

is a function of temperature only. The SI unit of pressure is the Pascal (Pa). 

INSTRUMENTATION 

The vapour pressure balance, assembled by the Department of Facilities Management at 

Huntingdon Life Sciences, consisted of an evaporator furnace, which contained a small 

aperture of fixed size in its roof. Immediately above the aperture, the scale pan of a CI 

Electronics microbalance was suspended. The balance and furnace are contained within a 

bell jar, so that the whole of the apparatus can be evacuated. 

CALIBRATION 

The microbalance was calibrated with a UKAS calibrated 1 mg weight (certified weight = 

1.0017 mg). During the calibration, the chart recorder was set to 5V full-scale (equivalent to 

180 mm on the chart recorder trace). The mean deflection observed using the 1 mg weight in 

triplicate determinations was 99 mm. It was found that 1 μg was equivalent to 2.745 x 10-3V. 

PROCEDURE 

A quantity of test substance (0.15 g) was added to the furnace, the apparatus assembled and 

evacuated to a pressure of less than 1 x 10-5 Torr (1.3 x 10-3 Pa). 

After stabilisation at a given temperature, the shutter was opened to allow a stream of vapour 

to impact upon one balance pan. The temperature and pan deflection were recorded on a 

chart recorder. The trace obtained enabled the calculation of mass difference. The furnace 

temperature was then raised in steps of approximately 5oC and further measurements taken. 

Two definitive runs were performed between temperatures of 89.5 and 125.0°C, the pressure 

being kept at less than 1 x 10-5 Torr (1.3 x 10-3 Pa) throughout. An initial run was 

disregarded, the high and variable results being indicative of degassing and/or removal of 

volatile impurities. Mass differences, condensation rates and the sample temperature were 

monitored on a chart recorder on every occasion that the shutter was opened. 
: 10 : 

CALCULATIONS 

Assuming no condensation, the vapour pressure is related to the observed mass difference by 

the relationship 

Vapour pressure = 

A

∆m • g

Equation 1 

When condensation occurs the mass difference can, by reduced momentum transfer, be 

decreased by up to a factor of two from that appropriate to equation 1. The vapour pressure 

can also be obtained from the kinematic theory relationship for effusion or complete 

condensation 

Vapour pressure = 

A

C

M

2 RT

•

π •

Equation 2 

The efficiency of the condensation may be determined from the relative magnitudes of the 

measured mass difference and the condensation rate. For a degree of condensation, α, the 

measured condensation rate must be divided by this factor (α) to obtain the maximum 

condensation rate to be used in equation 2. Likewise, the experimental mass differences must 

be modified by a factor of 2/(2-α), which gives the appropriate factor of two enhancement in 

the vapour pressure calculation when the molecules do not bounce back. 

By substituting equation 1 for vapour pressure in equation 2, and correcting for degree of 

condensation and enhancement, it is possible to calculate α at each data point: 

g

1

M

2 RT

2

2 -

 = 

C

m

•

•

•

∆ π

α

α

Equation 3 

Rearranging equation 3 gives equation 4 from which α can be calculated directly 

1

+ 0.5

2 RT

M

C

m.g

 =

−

































•

•

∆

π

α Equation 4 

The effect of α on the two methods for estimating vapour pressure yields identical results. 

Confidence in this system is demonstrated by the observation that the values of α approach, 

but never exceed, unity (when α=1 there is 100% condensation). 

The vapour pressure-temperature relationship is as follows: 

Log10Vp = 

Slope

T

+ Intercept Equation 5 

Consequently, a plot of log10Vp versus 1/T(K) should be linear, and by extrapolation the 

vapour pressure at 298.15K can be calculated. 
: 11 : 

Glossary of terms used in equations 1 to 5

A = Surface area of the aperture (5.952 x 10-6 m

2

) 

C = Maximum condensation rate (kg/s) 

g = Acceleration due to gravity (9.813 m/s2) 

M = Molar mass (kg/mol) 

∆m = Mass difference (kg) 

R = Universal gas constant (8.314 J/mol/K) 

T = Temperature (°C) + 273.15 (K) 

Vp = Vapour pressure (Pa) 

RESULTS 

The full results of the two definitive runs are presented in Tables 1 and 2, with a graphical 

representation of both data sets in Figure 1. 

Run 1 Run 2 

Correlation (r): -0.9952 -0.9989 

Slope: -7488 -8344 

Intercept: 16.94 19.11 

Vp (Pa) at 25°C: 6.6 x 10-9 1.3 x 10-9 

The two runs give a mean vapour pressure value at 25°C of 4 x 10-9 Pa. 

CONCLUSION 

The vapour pressure of was found to be 4 x 10-9 Pa at 25°C. 
: 12 : 

TABLE 1 

Determination of the Vapour Pressure of (Run 1) 

Temperature Mass Condensation Vapour pressure α Corrected vapour 

(°C) difference rate (Pa) pressure (Pa) 

(μg) (μg/s) From From From From 

Mass Condensation Mass Condensation 

 difference difference 

90.5 0.11 NC 0.00018 NC NC 0.00018 NC 95.0 0.20 0.010 0.00033 0.00041 0.765 0.00054 0.00054 

100.5 0.27 0.013 0.00045 0.00058 0.783 0.00074 0.00074 105.0 0.48 0.023 0.00078 0.00100 0.777 0.00128 0.00128 110.0 0.81 0.051 0.00133 0.00224 0.912 0.00245 0.00245 115.0 1.59 0.086 0.00262 0.00377 0.837 0.00451 0.00451 120.0 2.83 0.131 0.00467 0.00576 0.763 0.00755 0.00755 125.0 5.48 0.211 0.00904 0.00937 0.682 0.01373 0.01373 

NC = no condensation observed 
: 13 : 

TABLE 2 

Determination of the Vapour Pressure of (Run 2) 

Temperature Mass Condensation Vapour pressure α Corrected vapour 

(°C) difference rate (Pa) pressure (Pa) 

(μg) (μg/s) From From From From 

Mass Condensation Mass Condensation 

 difference difference 

89.5 0.07 NC 0.00012 NC NC 0.00012 NC 93.5 0.13 NC 0.00022 NC NC 0.00022 NC 99.5 0.24 0.009 0.00040 0.00040 0.669 0.00060 0.00060 

104.0 0.36 0.018 0.00060 0.00076 0.776 0.00098 0.00098 109.5 0.75 0.046 0.00123 0.00199 0.893 0.00223 0.00223 114.5 1.25 0.081 0.00207 0.00354 0.923 0.00384 0.00384 119.5 2.57 0.141 0.00424 0.00621 0.846 0.00734 0.00734 125.0 5.06 0.218 0.00834 0.00966 0.733 0.01317 0.01317 

NC = no condensation observed
: 14 : 

FIGURE 1 

Plot of log10Vp versus 1/T 

-4.50-4.00-3.50-3.00-2.50-2.00-1.50-1.00

0.00245 0.00250 0.00255 0.00260 0.00265 0.00270 0.00

1/T

Log10V

p

Run 1Run 2
: 15 : 

SURFACE TENSION 

 (EEC Method A.5 and OECD Guideline 115) 

METHOD 

The surface tension of a 90% saturated aqueous solution was determined at 20°C with a 

surface tension torsion balance using the OECD harmonised ring method. 

DEFINITION AND UNITS 

Surface tension (σ) is defined as the free surface enthalpy per unit of surface area and is 

reported in N/m (SI units) or mN/m (SI subunit). 

APPARATUS 

Surface tension torsion balance: White Electrical Instrument Co., Malvern Link, Worcs. 

Ring dimensions: Ring radius 6.37 mm 

Wire radius 0.150 mm 

Calibration: The calibration factor, øb, by which all instrument readings shall be multiplied, 

was determined in accordance with: 

øb =

 

o 

g

σ

σ

where 

σo = value cited in the literature for the surface tension of water (mN/m) at the test 

temperature 

σg = measured value of the surface tension of water (mN/m) at that test temperature 

Literature value for the surface tension of water at 20°C (σo) = 72.7 mN/m. 

Mean measured surface tension of water at 20°C (σg) = 72.5 mN/m. 

Therefore, the calibration factor, øb = 1.003 

TEST SOLUTION PREPARATION 

Test substance (approximately 70 mg) was combined with purified water (250 ml) and 

shaken at 30°C in a water bath for 3 days. After this period, the sample was equilibrated at 

20°C for 6 hours and then duplicate portions (approximately 75 ml) were centrifuged 

(2000 rpm, 5 minutes, 20°C). 

Aliquots (45 ml) of each clear supernatant were then diluted to 50 ml with purified water to 

produce 90% saturated aqueous solutions for measurement.
: 16 : 

PROCEDURE 

The test vessel was half filled with liquid, put on the tensiometer test platform, and raised so 

that the ring was 2-3 mm below the liquid surface. The platform was lowered to draw a 

lamella from the liquid surface at a rate of approximately 0.5 cm/min. The maximum force 

which arose just before the lamella was torn off (i.e. the uncorrected surface tension) was 

recorded. Measurements were made at 5-minute intervals until a constant value (to within 

0.5 mN/m) was recorded. 

RESULTS 

Test temperature: 20°C 

Solution concentration: 90% saturated 

Solution age: Prepared immediately before measurement 

Initial values for the surface tension were obtained as soon as possible after transfer of the 

aqueous solution to the measurement vessel. 

Time Surface tension (mN/m) 

(minutes) Measured Calibration corrected 

A B A B 

0 68.0 68.5 68.0 68.5 

5 68.0 68.0 68.0 68.0 

10 68.0 68.0 68.0 68.0 

15 68.0 68.0 68.0 68.0 

20 68.0 68.0 68.0 68.0 

25 68.0 68.0 68.0 68.0 

30 68.0 68.0 68.0 68.0 

From the results there was no apparent time dependence of surface tension after transfer to 

the measuring vessel. 

Overall mean calibration corrected surface tension = 68.0 mN/m (to the nearest 0.5 mN/m) 

Harkins-Jordan* corrected value = 0.00181x

2

 + 0.862x - 0.285 = 66.5 mN/m 

where x = overall mean calibration corrected surface tension 

CONCLUSION 

The surface tension of a 90% saturated aqueous solut was found to be 

66.5 mN/m. As the result is greater than 60 mN/m, considered to be 

surface active.

*HARKINS, W.D. and JORDAN, H.F., J. Amer. Chem. Soc. 52, 1751 (1930). 
: 17 : 

RELATIVE SELF-IGNITION TEMPERATURE FOR SOLIDS 

 (EEC Method A.16) 

METHOD 

The presence or otherwise of a self-ignition temperature up to the melting temperature of the 

test substance was determined through the monitoring of the sample temperature while 

located in a steadily heated oven. 

DEFINITION AND UNITS 

The self-ignition temperature is defined as the minimum ambient temperature, expressed in 

degrees Celcius (°C), at which a certain quantity of a substance will ignite when mixed with 

air under defined conditions. In accordance with the Directive, the self-ignition temperature 

in this test will be taken as the oven temperature at which the sample temperature reaches 

400°C by self-heating. 

APPARATUS 

Carbolite furnace with natural air circulation and explosion relief. 

Standard K-type thermocouples for temperature measurement. 

Open topped 20 mm cube made from stainless steel wire mesh with 0.045 mm openings. 

PICO ADC16 data logger with PICOLOG data logging software. 

Metal base plate. 

Calibration: The furnace controller display is regularly calibrated against a standard 

thermometer. Instrument readings collected by the software are scaled from data 

points manually entered. 

PROCEDURE 

The mesh cube was filled with test substance by tapping gently and adding more until the 

cube was full. The cube was then suspended in the centre of the furnace at room temperature 

and a thermocouple placed in the centre of the cube. Another thermocouple was placed 

midway between the furnace wall and the cube in order to monitor the furnace temperature. 

The temperature of the furnace and sample were then continuously recorded while the 

temperature of the furnace was increased at a rate of 0.5°C/min to 400°C. 

RESULTS 

There was no exothermic reaction of below its melting point at approximately 

150°C.

The temperature versus time graph is presented in Figure 2. 

CONCLUSION 

 did not self-ignite below its melting temperature. 
: 18 : 

FIGURE 2 

Temperature Versus Time Curve 

0

50

100150200250300350400

0 100 200 300 400 500 600 

Time (minutes)

Temperature (°C) Sample

Oven
: 19 : 

PARTICLE SIZE DISTRIBUTION 

METHOD 

The particle size distribution was initially examined using sieve analysis. As greater than 

10% by weight of the test substance was found to pass a 75 micron sieve, it was further 

examined by image analysis. 

PROCEDURE 

Sieve analysis 

A portion (10 g) of the test substance was weighed into the coarsest of a nest of sieves 

(100 mm diameter) of aperture sizes 400, 125, 75, 30 and 10 microns. The nest was shaken 

for 30 minutes with tapping (Pascall Inclyno sieve shaker), and the proportion of the test 

material passing through each sieve determined gravimetrically. The test was performed in 

duplicate, and the results are summarised thus: 

Particle size % in range by weight 

(μm) Test 1 Test 2 Mean 

>400 

400 - 125 

125 - 75 

75 - 30 

30 - 10 

<10 

Image analysis (based on BS3406, Part 4, 1993)

A suitable suspending medium was experimentally determined to fulfil the following 

properties. 

i) low solubility 

ii) different refractive index 

iii) low volatility 

iv) good dispersion 

Screening to remove particles greater than 125 μm was not considered necessary, partly due 

to the relatively small proportion (less than 10% by mass) of the test substance found to be 

greater than this size in the sieve analysis test, but primarily due to the observations made on 

microscopic examination of an initial slide. This microscopic examination showed the 

particles to be predominantly long and thin, with none noticeably greater than 125 μm in 

width. It was considered that the proportion of particles retained on the 125-μm sieve in the 

sieve analysis test was influenced by the alignment of the particles on the sieve and that this 

proportion would have decreased on extended sieving. 
: 20 : 

Six replicate samples, each approximately 5-10 mg in weight, were suspended by agitation 

and mounted on a microscope slide for image analysis.

APPARATUS 

The following conditions were employed: 

Microscope: Leitz Laborlux S 

Video monitoring: Ikegami charge coupling device (CCD) camera, model 

1CD-42E, with monitor (NEC/Multisync 3D) 

Illumination type: Kohler 

Suspending medium: 1% v/v aqueous Tween 80 

Size range: 0.5 - 600 μm 

Magnification: X10, X40 

Size parameters: Longest dimension, axial ratio 

RESULTS 

The reported results were prepared from combined data for both magnifications, with at least 

50 fields counted at each magnification. The mean particle length and axial ratio are used to 

calculate particle volumes in each range which, when combined with the particle number 

distribution, give a volume or mass distribution. 

The results can be summarised as follows: 

Particle size % Mass 

(μm) Test 1 Test 2 Test 3 Test 4 Test 5 Test 6 Mean 

320 - 600 

160 - 320 

60.0 - 160 

30.0 - 60.0 

10.4 - 30.0 

0.5 - 10.4 

CONCLUSION 

The results indicate that 0% w/w of is smaller than 10 μm. 
: 21 : 

SOIL ADSORPTION 

 (EEC Method C.19 and OECD Guideline 121) 

METHOD 

The sorption behaviour of the test substance on soil was examined using high performance 

liquid chromatography (HPLC). 

HPLC is performed on commercially available analytical columns packed with a solid phase 

containing a moderately polar stationary phase with lipophilic and polar moieties (e.g. cyanopropyl bonded phase). 

Chemicals injected onto such a column move along it by partitioning between the mobile 

phase and the stationary phase. The velocity of each component thereby depends on the 

degree of adsorption on the stationary phase. The dual nature of the stationary phase allows 

for interaction of polar and apolar parts of a molecule in a similar way as is the case for soil. 

This enables the relationship between the retention time on such a column and the adsorption 

coefficient on the organic parts of the soil to be established. 

The adsorption coefficient is deduced from the capacity factor (k’) given by: 

k’ = 

0

r 0

t

t - t

where tr is the measured retention time of sample or reference; and 

 t0 is the retention time of the internal deadtime standard. 

DEFINITION AND UNITS 

The adsorption coefficient, Koc, is defined as the distribution coefficient (Kd) normalised to 

the organic carbon content of a sorbent. Kd is defined as the ratio between the concentration 

of the test substance in the soil and the concentration of the substance in the aqueous phase, 

both measured when adsorption equilibrium is reached. 

The adsorption coefficient is usually given in the form of the logarithm to base ten (log10Koc). 

PROCEDURE

In order to correlate the measured capacity factor (k’) of the test material with its Koc, six 

reference materials were selected from a list of recommended compounds. These reference 

compounds were selected to have log10Koc values between 1.3 and 5.6. An internal deadtime 

standard (I.D.S., formamide) with no retention on the HPLC column was used to determine 

the deadtime of the HPLC system. 

 
: 22 : 

The following reference mixture was prepared in HPLC mobile phase. 

Reference substance Known* Concentration 

log10Koc (mg/l) 

Formamide (I.D.S.) Deadtime only 1050 

Acetanilide 1.3 54.0 

Nitrobenzene 2.4 49.9 

Naphthalene 2.8 50.6 

2-Methylnaphthalene 3.9 62.2 

Anthracene 4.4 47.3 

DDT 5.6 107 

*values from OECD Method 121, with the exception of nitrobenzene, 2-methylnaphthalene and 

anthracene, values for which came from ‘HODSON, J. and WILLIAMS, N.A., (1988). The 

Estimation of the Adsorption Coefficient (Koc) for Soils by High Performance Liquid 

Chromatography, Chemosphere, Vol. 17, pp.67-77, Pergammon Journals Ltd’. 

A solution of (52.0 mg/l in mobile phase, and containing I.D.S. at the same 

concentration a erence mixture) was prepared and chromatographed in duplicate, 

bracketed by samples of the reference mixture, using the HPLC conditions below. 

HPLC CONDITIONS 

Instrument: 

Column: 

Column temperature: 

Mobile phase composition: 

Flow rate: 

Injection volume: 

Detector: 
: 23 : 

RESULTS

Table 3 presents the primary data from which the calibration curve (Figure 3) was generated. 

Figures 4 and 5 present examples of chromatograms generated during this test. 

The mean calibration curve was used to interpolate the results. 

The mean calibration curve produced was: 

log10Koc = 3.46 log10k’ + 3.86 

where k’ = capacity factor 

 = 

0

r 0

0

r

t

t - t

 = 

t

t'

and t'r = true retention time obtained by subtracting deadtime, or t0, of I.D.S. from measured 

retention time of sample or reference standard. 

A mean log10Koc value for of 3.7 was thus obtained. 

A ranking system of soil mobility based upon Koc has been proposed by McCall et al

(Measurement of Sorption Coefficients of Organic Chemicals and their use in Environmental 

Fate Analysis, Test Protocols for Environmental Fate & Movement of Toxicants, Proceedings 

of Association of Official Analytical Chemists, 1980): 

Koc Mobility class 

0 - 50 very high 

50 - 150 high 

150 - 500 medium 

500 - 2000 low 

2000 - 5000 slight 

> 5000 immobile 

The determined log10Koc value of 3.7 (Koc > 5000) suggests that the test substance will be 

immobile in soil. 

CONCLUSION 

 was found to have a log10Koc value of 3.7. 
: 24 : 

TABLE 3 

HPLC Determination of log10Koc for 

Sample Component tr (minutes) k' log10k' 

Standard Run A Acetanilide 3.797 0.169 -0.773 (t0 = 3.249 minutes) Nitrobenzene 4.485 0.380 -0.420 

Naphthalene 5.392 0.660 -0.181 

2-Methylnaphthalene 6.000 0.847 -0.072 

Anthracene 7.985 1.458 0.164 

DDT 13.185 3.058 0.485 

Sample A 6.186 0.906 -0.043 (t0 = 3.246 minutes) Sample B 6.180 0.904 -0.044 (t0 = 3.246 minutes) Standard Run B Acetanilide 3.790 0.168 -0.774 (t0 = 3.244 minutes) Nitrobenzene 4.475 0.379 -0.421 

Naphthalene 5.374 0.657 -0.183 

2-Methylnaphthalene 5.977 0.842 -0.074 

Anthracene 7.946 1.449 0.161 

DDT 13.108 3.041 0.483 

Linear regression (based on mean of standard runs A and B): log10Koc = 3.46 log10k’ + 3.86
: 25 : 

FIGURE 3 

log10k' Versus log10Koc Calibration of Standard Runs A and B 

log10Koc = 3.46 log10k' + 3.86

r = 0.9877

0.001.002.003.004.005.006.007.00

-1.000 -0.800 -0.600 -0.400 -0.200 0.000 0.200 0.400 0.600 

log10k'

log10Koc
: 26 : 

FIGURE 4 

Representative Chromatogram of Standard Mix with log10Koc between 1.3 

0 2 4 6 8 10 

mAU

0

200400600800

10001200

 VWD1 A, Wavelength=210 nm (0SA_00F\RSPSYS02.D)

 3.249

 3.797

 4.485

 5.392

 6.000

 7.985
: 27 : 

FIGURE 5 

Representative Chromatogram of 
: 28 : 

DEVIATIONS FROM PROTOCOL 

The final paragraph of Section 2.4.3 of the protocol states that samples will be ‘screened to 

remove particles of size greater than 125 μm’ for image analysis. This was not done, the 

reasons for which are given on Page 19 of the report. 

MAINTENANCE OF RECORDS 

All raw data, samples and specimens (if appropriate) generated at Huntingdon Life Sciences 

during the course of this study, together with a copy of the final report, will be retained by 

Huntingdon Life Sciences in its archive for a period of five years from completion of the 

study, or until the Sponsor provides instructions for their delivery to the Sponsor’s designated 

archiving facility. After such time, the Sponsor will be contacted and advice sought on the 

return, disposal or further retention of the materials. Types of sample and specimen which are 

unsuitable, by reason of instability, for long term retention and archiving may be disposed of 

after the periods stated in Huntingdon Life Sciences Standard Operating Procedures and after 

contacting the Sponsor. Under no circumstances will any item be discarded without the 

Sponsor’s consent. 

Huntingdon Life Sciences will retain the Quality Assurance records relevant to this study and 

a copy of the final report in its archive indefinitely. 
: 29 : 

REFERENCES 

The UK Good Laboratory Practice Regulations (Statutory Instrument 1999 No. 3106, as 

amended by Statutory Instrument 2004 No. 994). 

OECD Principles of Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM(98)17. 

EC Commission Directive 2004/10/EC of 11 February 2004. 

Directive 92/69/EEC, Official Journal No. L383A, 29.12.1992, Method A.4, Vapour 

Pressure. 

OECD Guideline 104 - Vapour Pressure, Adopted by the Council on 23 March 2006. 

Directive 92/69/EEC, Official Journal No. L383A, 29.12.1992, Method A.5, Surface 

Tension. 

OECD Guideline 115 - Surface tension, Adopted by the Council on 27 July 1995. 

Directive 92/69/EEC, Official Journal No. L383A, 29.12.1992, Method A.16, Relative 

Self-Ignition Temperature for Solids. 

Directive 2001/59/EEC, Official Journal No. L225, 21.8.2001, Method C.19, Estimation of 

the Adsorption Coefficient (Koc) on Soil and on Sewage Sludge Using High Performance 

Liquid Chromatography (HPLC). 

OECD Guideline 121 - Estimation of the Adsorption Coefficient (Koc) on Soil and on Sewage 

Sludge Using High Performance Liquid Chromatography (HPLC), Adopted by the Council 

on 22 January 2001. 

McCall et al, Measurement of Sorption Coefficients of Organic Chemicals and their use in 

Environmental Fate Analysis, Test Protocols for Environmental Fate & Movement of 

Toxicants, Proceedings Of Association of Official Analytical Chemists, 1980.
: 30 : 

APPENDIX 1 

EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2005
: 31 : 
: 32 : 

APPENDIX 2 

CERTIFICATE OF ANALYSIS
: 33 : 
: 34 : 

APPENDIX 3 

STUDY PROTOCOL
: 35 : 
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: 50 : 
: 51 : 
: 52 : 
: 53 : 
: 54 :