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### 2 - Bromo - 3,3,3 - Trifluoropropene P hysicochemical P roperties

HLS study number: WA G0005 Version ID: Final Issue date: 12 July 2012Huntingdon Life Sciences WA G0005

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## Sponsor and Te st Facility Sponsor American Pacific Corporation 3883 Howard Hughes Parkway Sui te 7 00 Las Vegas NV 89169 USA Test facility Hunt ingdon Life Sciences Eye Research Centre Eye Suffo lk IP23 7PX UKHuntingdon Life Sciences WA G0005

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## Ta ble of Content

s Sponsor and Test Facility ................................ ................................ ................................ ...... 2 Compliance w ith Good Laboratory Practice ................................ ................................ .......... 6 Quality Assurance Statement ................................ ................................ ................................ 7 Contributing Scientists ................................ ................................ ................................ .......... 8 Summary ................................ ................................ ................................ .............................. 9 1. Introduction ................................ ................................ ................................ ................ 10 1.1 Objective ................................ ................................ ................................ ......... 10 1.2 Regulatory compliance ................................ ................................ ................... 10 1.3 Study schedule ................................ ................................ ............................... 10 1.4 Maintenance of records ................................ ................................ .................. 10 2. Test Substance ................................ ................................ ................................ .......... 11 3. Experimental Procedures ................................ ................................ ........................... 12 3.1 Freezing temperature ................................ ................................ ...................... 12 3.1.1 Method ................................ ................................ ................................ 12 3.1.2 Definition and units ................................ ................................ .............. 12 3.1.3 App aratus ................................ ................................ ............................ 12 3.1.4 Procedure ................................ ................................ ........................... 12 3.1.5 Results ................................ ................................ ................................ 12 3.1.6 Conclusion ................................ ................................ .......................... 12 3.2 Boili ng temperature ................................ ................................ ......................... 13 3.2.1 Method ................................ ................................ ................................ 13 3.2.2 Definition and units ................................ ................................ .............. 13 3.2.3 Apparatus ................................ ................................ ............................ 13 3.2.4 Procedure ................................ ................................ ........................... 13 3.2.5 Results ................................ ................................ ................................ 14 3.2.6 Conclusion ................................ ................................ .......................... 14 3.3 Relative density ................................ ................................ .............................. 15 3.3.1 Method ................................ ................................ ................................ 15 3.3.2 Definition and units ................................ ................................ .............. 15 3.3.3 Apparatus ................................ ................................ ............................ 15 3.3.4 Reagent s ................................ ................................ ............................. 15 3.3.5 Procedure ................................ ................................ ........................... 15 3.3.6 Results ................................ ................................ ................................ 15 3.3.7 Conclusion ................................ ................................ .......................... 15Huntingdon Life Sciences WA G0005

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## Ta ble of Content

s - continued 3.4 Vapour pressure ................................ ................................ ............................. 16 3.4.1 Method ................................ ................................ ................................ 16 3.4.2 Definition and units ................................ ................................ .............. 16 3.4.3 Apparatus ................................ ................................ ............................ 16 3.4.4 Procedure ................................ ................................ ........................... 16 3.4.5 Calculations ................................ ................................ ........................ 17 3.4.6 Results ................................ ................................ ................................ 17 3.4.7 Conclusion ................................ ................................ .......................... 17 3.5 Surface tension ................................ ................................ ............................... 22 3.5.1 Method ................................ ................................ ................................ 22 3.5.2 Definition and units ................................ ................................ .............. 22 3.5.3 Apparatus ................................ ................................ ............................ 22 3.5.4 Test solution preparation ................................ ................................ ..... 22 3.5.5 Procedure ................................ ................................ ........................... 23 3.5.6 Results ................................ ................................ ................................ 23 3.5.7 Conclusion ................................ ................................ .......................... 23 3.6 Wa ter solubility ................................ ................................ ............................... 24 3.6.1 Definition and units ................................ ................................ .............. 24 3.6.2 Preliminary test ................................ ................................ ................... 24 3.6.3 Procedure for the definitive test ................................ ........................... 24 3.6.4 HPLC conditions ................................ ................................ ................. 24 3.6.5 Preparation of calibration ................................ ................................ ..... 25 3.6.6 Calculations ................................ ................................ ........................ 25 3.6.7 Results and discussion ................................ ................................ ........ 25 3.6.8 Conclu sion ................................ ................................ .......................... 25 3.7 Partition coefficient ................................ ................................ ......................... 29 3.7.1 Definition and units ................................ ................................ .............. 29 3.7.2 Preliminary estimation ................................ ................................ ......... 29 3.7.3 Method ................................ ................................ ................................ 29 3.7.4 Procedure ................................ ................................ ........................... 29 3.7.5 HPLC conditions ................................ ................................ ................. 30 3.7.6 Results ................................ ................................ ................................ 30 3.7.7 Conclusion ................................ ................................ .......................... 30 3.8 Flash point ................................ ................................ ................................ ...... 35 3.8.1 Method ................................ ................................ ................................ 35 3.8.2 Definition and units ................................ ................................ .............. 35 3.8.3 Apparatus ................................ ................................ ............................ 35 3.8.4 Procedure ................................ ................................ ........................... 35 3.8.5 Calculation ................................ ................................ .......................... 35 3.8.6 Results ................................ ................................ ................................ 36 3.8.7 Conclusion ................................ ................................ .......................... 36 3.9 Explo sive properties ................................ ................................ ....................... 37 3.9.1 Assessment of explosive properties ................................ .................... 37 3.9.2 Conclusion ................................ ................................ 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## Ta ble of Content

s - continued 3.10 Auto - ignition temperature (liqu ids and gases) ................................ ................. 38 3.10.1 Method ................................ ................................ ................................ 38 3.10.2 Definition and units ................................ ................................ .............. 38 3.1 0.3 Apparatus ................................ ................................ ............................ 38 3.10.4 Procedure ................................ ................................ ........................... 38 3.10.5 Results ................................ ................................ ................................ 38 3.10.6 Conclusion ................................ ................................ .......................... 38 3.11 Oxidi sing properties ................................ ................................ ........................ 39 3.11.1 Assessment of oxidising properties ................................ ..................... 39 3.11.2 Conclusion ................................ ................................ .......................... 39 3.12 Soil adsorption ................................ ................................ ................................ 40 3.12.1 Definition and units ................................ ................................ .............. 40 3.12.2 Method ................................ ................................ ................................ 40 3.12.3 Procedure ................................ ................................ ........................... 40 3.12.4 HPLC conditions ................................ ................................ ................. 41 3.12.5 Results ................................ ................................ ................................ 42 3.12.6 Conclusion ................................ ................................ .......................... 42 

## List of A nnexes Annex 1 Eye Research Centre GLP Compliance Statement 2011 ............................. 47Huntingdon Life Sciences WA G0005

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## Contributing S cientists 2 - Bromo - 3,3,3 -trifluoropropene : Physicochemical properties Study management P Sy dney BSc MSc Study Di rector Product Chemistry Other responsible staff L Lawson MChem Sci ent ist Product Chemistry S Cage HNC Sci ent ist Product Chemistry S Fox S ci ent ist Product Chemistry A Keep Sci ent ist Product ChemistryHuntingdon Life Sciences WA G0005

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## Summary A study w as perf ormed to determine so me physicochemical properti es of the test substance, 2 - Bromo - 3,3,3 - trifluoropropene . The methods followed are amongst those described in Commissio n Regulat io n (EC) No. 440/2008 and the OECD Guidelines fo r the Testing of Chemicals. T he physicochemical properti es which have been determined in this study are detailed below, together with the result obtained for each test . E C OECD Test Result method no. method no. A . 1 102 Freezing tem perature Less than - 50 °C A . 2 103 Boiling temperature 34  C A . 3 109 Relative density ) (D 20 4 1. 65 A . 4 104 Vapour pressure 82000 Pa at 25°C A . 5 115 Surface tension 72.0 mN/ m (1 g/l solution) A . 6 105 Water solubility 1.0 g/l A . 8 1 1 7 Partition coefficient Log 10 P ow = 2.7 A . 9 - Fla sh point None determined before the sample boiled A . 14 - Explosive properties (theoretical assessment) Not explosive A . 1 5 - Auto - ignition temperature (liquids and gases) None below 400  C A . 21 - Oxidising properties (theoretical assessment) Non - oxidising C . 19 121 Soil adsorption Log 10 K oc = 2.2Huntingdon Life Sciences WA G0005

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## 1. Introduction 1.1 Objective This study

was designed to determine some physicochemical properti es of the test substance , 2 - Bromo - 3,3,3 - trifluoropropene . I nformation on physicochemical properti es is important in the assessment of the potential effects of a substance both at work and in the environment. 

### 1.2 Regulatory compliance The study

w as conducted in accordance wi th a ccepted principles in order to meet the requi rements of Commissio n Regulat io n (EC) No. 440/2008 and the OECD Guidelines fo r the Testing of Chemicals. 

### 1.3 S tudy schedule Study init iat io n d ate : 09 November 2011 Exper imental start d at e : 28 November 2011 Experimental c om

plet io n d ate : 22 March 2012 Study c om

plet io n d ate : 12 July 2012 

### 1.4 Maintenance of records Primary data fro m the test s , and a copy o f the final report, are stored in the archives of Hunt ingdon Life Scienc es.Huntingdon Life Sciences WA G0005

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## 2. Te st S ubstance Ident it y: 2 - Bromo-3,3,3 - trifluoropropene Chemical s tructure: F F F CH 2

Br CAS number: 1514 -82 -5 Appearance: Clear liquid wi th a slight yellow tint Storage condit io ns: Room tem perature Lot nu mber: Kg F080 Puri ty : 99.9 % excluding stabilisers Stabilisers:

Expi ry date: 01 August 2 012 Date received: 12 September 2011Huntingdon Life Sciences WA G0005

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## 3. Experimental Procedure s 3.1 Freezing t emperature (E C Method A . 1, OECD Method 102) 3.1.1 Method The freezing point was determined in a sui table apparatus in accordance wi th Bri ti sh Standard Method BS4633 - Method for the Determinat io n of Crystallizing Point (1970). 3.1.2 Definition and units The freezing tem

perature of a substance is defined as the tem perature (°C) at which the phase transi tion f rom l iquid to solid state, at normal at mo spheric pressure, takes place. 3.1.3 Apparatu s Seta Freezing Po intApparatus, Stanhope Seta Ltd. Calibrat io n: The apparatus i s regularly calibrated by determination of the freezing points of

reference materials. 3.1.4 Procedure The Dewar flask was filled wi th a f reezing mixture of

a cetone/so lid carbon dio xide and the sample tube filled to a sui ta ble depth. The sam

ple w as st irred cont inuously and the tem perature was measured at intervals. 3.1.5 Results T he test substance did not solidify as the temperature was decreased fro m ambient tem perature down to - 50 °C. T h e sample was still liquid at - 50 °C , which is the minimu m tem perature achievable with the apparatus . 3.1.6 Conclusion The freezing tem

perature of 2 - Bromo - 3,3,3 - trifluoropropene was determined to be less than

- 50 °C.Huntingdon Life Sciences WA G0005

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### 3.2 Boiling temperature (E C Method A . 2, OECD Method 103) 3.2.1 Method The bo iling temperature was determined by the Siwolobo ff method. 3.2.2 Definition and units The standard bo iling tem perature is defined as the tem perature (°C) at which the pressure of the saturated vapour of a liquid is equal to the standard press ure. 3.2.3 Apparatus Mel ting/bo iling point apparatus: Model B - 545, Buchi. Calibrat io n: The apparatus is regularly calibrated by determination of the bo iling points of reference materials. 3.2.4 Procedure A bo iling tube (3.2 mm diameter) was filled to a height of ap proximately 5 to 10 mm

wi th the sample and a bo iling capillary was immersed, open end first. The bo iling tube containing the sample and the boiling capillary was inserted into the sam ple ho lder and the tem

perature was raised to approximately 10°C below the ant ic ipated bo iling point. The tem perature was then raised at approximately 1°C/min and the tem perature noted at which a rapid cont inuous stream of bubbles was seen emerging from the inverted open end of the capillary . This tem

perature represents the boil ing point.Huntingdon Life Sciences WA G0005

14 3.2.5 Results The results o f duplicate boiling point determinat io ns on 2 - Bromo - 3,3,3 - trifluoropropene were as fo llo ws: Barometric pressure: 10 25 mbar Boiling point (  C) Observed Pressure corrected * Run 1 34.7 34.4 Run 2 34.6 34.3 *Pres sure correction C b = T b + 0.00009 (1013 - P b )(273 + T b ) where C b is the corrected boiling point (oC) P b is the ambient baro met ric pressure (mbar) and T b is the observed bo iling point (  C) 3.2.6 Conclusion The bo iling tem

perature of 2 - Bromo - 3,3,3 - trifluoropropene was determine d to be 34 .4 °C at 1013 mbar (1 atm) .Huntingdon Life Sciences WA G0005

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### 3.3 Relative density (E C Method A . 3, OECD Method 109) 3.3.1 Method The relat ive densit y of

2 - Bromo - 3,3,3 - trifluoropropene was determined relat ive to purified water using a densit y meter at 20  C. 3.3.2 Definition and units The relat ive densi

ty (D 4 T ) of solids and liquids is defined as the ratio of the mass of

a v ol ume of substance to be examined, determined at T°C, and the mass of the same volume of water at 4°C. 3.3.3 Apparatus Densit y meter: Model DA - 100M, Mettler Toledo 3.3.4 Reagents Water: Purified by reverse osmosis and deionising 3.3.5 Procedure Test tem perature: 20°C The instrument calibrat io n was verified by measurement of p urified water att he test tem perature. The test substance was then int roduced to the oscillator tube in the densi ty m eter via syringe and the densit y recorded when the value had stabilised. The test was performed in duplicate. 3.3.6 Results The densit y of

w ater at the test tem

perature was determined to be 0.998 g/ml , in good agreement wi

th t he known value of

0 .998 g/ ml , th

us confirming correct calibrat io n of t he instrument. Subsequently, duplicate measurements of t he densit y of

t he test substance of 1.653 g/ ml and 1.653 g/ml were recorded, yielding a me an value of 1.653 g/ml . This is equivale ntt o a relat ive densit y value of 1.653 (relat ive to the densit y of water at 4  C). 3.3.7 Conclusion The relat ive densit y )

(D 20 4 of 2 - Bromo - 3,3,3 - trifluoropropene was f ound to be 1.65 .Huntingdon Life Sciences WA G0005

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### 3.4 Vapour pressure (E C Method A . 4, OECD Method 104) 3.4.1 Method The vapour pressure of 2 - Bromo - 3,3,3 - trifluoropropene was determined at tem peratures between 2 73 and 293 K ( 0 and 20  C) by the static measurement procedure . 3.4.2 Definition and units The vapour pressure of

a s ubstance is defined as the saturation pressure above a so lid o r liquid substance. At the therm odynamic equilibrium, the vapour pressure of a pure substance is a funct io n of tem

perature only . The SI uni t of pressure i s the Pascal (Pa). 3.4.3 Apparatus Static vap our pressure apparatus (Figure 1 ) Mano met er fluid: mercury Pu mp ing system: high vacuum pump, Edwards 3.4.4 Procedure Leak rate determination : Before use for vapour pressure determination, the leak rate of the apparatus was checked. The apparatus was evacuated to minimum pressure at room t em perature wit h the sam

ple iso lation valve V 1 in the open posi tion (no sam

ple present). The leak rate was then determined by closing V 2 , di scontinuing the vacuum pumping, and measuring the increase in pressure over a specified period. The leak rate was fo und to be suffic iently lo w ( 0.5 P a per minute) to allow the test to proceed. Vapour pressure determinat io n : The sample container was approximately half filled wi th 2 - Bromo - 3,3,3 - trifluoropropene, attached to the apparatus and cooled wi thl iquid nit rogen. The apparatus was then evacuated to minimum pressure, wi th t he fine control

v alve (FCV) closed and the iso lat io n valves V 1 and V 2 open, in order to degas the sample. Valves V 1 and V 2 were then simultaneously clo sed and the sample container introduced into a constant tem

perature water bath , the vacuum pumping having been discontinued. The init ia l pressure of the system was recorded. After being allowed to equilibrate at test temperature for 10 minutes, the resul

ting displacement of

t he mano met er fluid was then adj usted by the int roduction of

a irt hrough the fine control v alve unt il the mano met er balance was restored. The pressure was again recorded. Two runs were performed between temperatures of 2 73 and 293 K ( 0 and 20  C) . The perf ormance of

t he apparatus was checked by measurement of t he ref erence substance, acetone .Huntingdon Life Sciences WA G0005

17 3.4.5 Calculations The vapour pressure, Vp (Pa), of the sample at each test temperature was calculated thus:

Vp (Pa) = Pressure balanced (Pa) - Pressure unbalanced (Pa) The vapour pressure of 2 - Bromo - 3,3,3 - trifluoropropene at 298K was extrapolated f rom a plot of log 10 Vp against 1/T. 3.4.6 Results A measured value of 32000 Pa at 25  C was obtained fo r the vapour pressure of the reference substance , acetone , in good agreement wit h the literature value of 30800 Pa ( LIDE, D.R. , Handbook o

f Chemistry and Physics , CRC Press, 75 th Edi tion, 1994 ). The fu ll resul

ts of the two runs fo r 2 - Bromo - 3,3,3 - trifluoropropene are presented in Tables 1 and 2, wi th a graphical representation in Figure 2. Run 1 Run 2

Correlation: - 0.9993 - 0.9999 Slop e: - 1380 - 1427 Intercept: 9.539 9.700 Vp (Pa) at 25°C: 81000 82000 The data gave a mean vapour pressure val ue of 68000 Pa at 20°C. Linear extrapolation of the data results in 82000 Pa at 25oC . 3.4.7 Conclusion The test substance, 2 - Bromo - 3,3,3 - trifluoroprope ne , was fo und to have a vapour pressure of 82000 Pa at 25°C .Huntingdon Life Sciences WA G0005

18 Figure 1 Static vapour pressure apparatus

Key: A Pressure gauge B Vacuum C Sam

ple D Manometer fluid E Air FCV Fine control valve V 1 , V 2 Isolation valves19Huntingdon Life SciencesWAG0005

Tabl e 1 Determination of the vapour pressure of 2 - Bromo - 3,3,3 -trifluoropropene (Run 1) Temperature Temperature 1/Temperature Vapour pressure Log 10 (°C) (K) (1/K) (Pa) vapour pressure 0 273 0.00366 31000 4.49 5 278 0.003 60 3700 0 4.57 10 283 0.003 53 46000 4.66 15 288 0.00347 56000 4.75 20 293 0.00341 68000 4.8320Huntingdon Life SciencesWAG0005

Table 2 Determination of the vapour pressure of 2 - Bromo - 3,3,3 -trifluoropropene (Run 2) Temperature Temperature 1/Temperature Vapour pre ssure Log 10 (°C) (K) (1/K) (Pa) vapour pressure 0 273 0.00366 30000 4.48 5 278 0.003 60 37000 4.57 10 283 0.003 53 46000 4.66 15 288 0.00347 56000 4.75 20 293 0.00341 68000 4.8321Huntingdon Life SciencesWAG0005

Figure 2 Plot of log 10 Vp versus 1/T for 2 - Bromo - 3,3,3 - trifl uoropropene

4.45 4.50 4.55 4.60 4.65 4.70 4.75 4.80 4.85 4.90

0.00335 0.00340 0.00345 0. 00350 0.00355 0. 00360 0.00365 0. 00370Log10Vp

1/K Ru n 1 Ru n 2Huntingdon Life Sciences WA G0005

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### 3.5 Surface tension (EC Method A.5, OECD Method 115) 3.5.1 Method The surface tension

of a 1 g/l aqueous sol

ut io n was determined wi

th a surf ace tension

torsion balance using the OECD harmo nised ring method. 3.5.2 Definition and units Surface tension

( ) is defined as the free surface enthalpy per uni t of surface area and is reported in N/m (SI units) or mN/m (SI subunit). 3.5.3 Apparatus Surface tensio n torsion balance: Torsion Balance fo r Surf ace and Interfacial Tension Measurement, White Electrical Instrument C o., Malvern Link, Worcs. Ring dimensio ns: Ring radius 6.37 mm Wire radius 0.150 mm Calibrat io n: The calibrat io n factor, ø b , by which all instrument readings shall be m ul tiplied, was determined in accordance with:

ø b = o g   where  o = value ci

ted in the literature fo r the surface tension of w ater (m

N/m) at the test tem perature  g = measured value of the surface tensio n of water (mN/m) at that test temperature Li terature val

ue f or the surface tensio n of water at 20  C (  o ) = 72.7 m N/m . Measured surface tensio n of water at 20  C (  g ) = 72 .0 mN/ m. Therefore, the calibration factor, ø b = 1.01 3.5.4 Test solution preparation Test substance (approximately 50 mg ) was added to purified water (5 0 ml) in a vo lu metric flask and the sample was tight ly sealed and ul trasonicated fo r 30 minutes unt il the sample was dissolved , before re - equilibrat io n at 20oC and measurement. Duplicate samples were prepared in this manner.Huntingdon Life Sciences WA G0005

23 3.5.5 Procedure The test vessel was half filled wi th l iquid, put on the tensiom eter test platform, and raised so that the ring was ju st below the liquid surface. Th e plat fo rm w as lo wered to draw a la mella fro m the liquid surface. The maximum force which arose just before the lamella was torn off is the surface tension

a nd was recorded. Measurements were made at int ervals until a constant value (to within 0.5 mN/m) was recorded. 3.5.6 Results Test tem perature: 20°C Sol ution concentration: 1 g/l Sol ution age: Prepared immediately before measurement Ini ti al v alues fo r the surface tens io n were obtained as soon as possible af ter transfer of

t he aqueous sol

ution to the measurement vessel. Time Surface tension (mN/m) (minutes) Measured Calibration corrected A B A B 0 72.0 72.5 72.7 73.2 5 72.0 72.0 72.7 72.7 10 72.0 72.0 72.7 7 2.7 15 72.0 72.0 72.7 72.7 20 72.0 72.0 72.7 72.7 25 72.0 72.0 72.7 72.7 30 72.0 72.0 72.7 72.7 Overall mean calibrat io n corrected surface tensio n = 73.0 mN/ m Harkins - Jordan* corrected value = 72 .0 mN/m 3.5.7 Conclusion The surface tension

of a 1 g/l aqueous sol

ut io n of 2 - Bromo - 3,3,3 - trifluoropropene was fo und to be 72 .0 mN/ m. A s the resul t was greater than 60 mN/m, the test substance was not considered to be surface act ive.

*HARKINS, W.D. and JORDAN, H.F., J. Amer. Chem. Soc. 52 , 1751 (1930).Huntingdon Life Sciences WA G0005

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### 3.6 W ater solubility (EC Method A.6, OECD Method 105) 3.6.1 Definition and units The so lubilit y in water is specified by the saturation mass concentration of t he substance in water, and is a funct io n o f tem perature. Sol

ubili tyi s specified in uni

ts of m ass per vol

um e of sol ut io n, and is reported as grams/litre (g/l). 3.6.2 Preliminary test A preliminary test was conducted by shaking a known mass (approximately 10 0 mg) of t he test substance wi

th increasing vo lu mes of purified water and visually assessing disso lut io n. The so lubilit y of 2 - Bromo - 3,3,3 - trifluoropropene was estimated to be between 200 and 1000 mg/ l and a definit ive test was subsequently performed by the flask method . 3.6.3 Procedure for the definitive test Wheaton vials containing approximately 300 m g of 2 - Bromo - 3,3 ,3 - trifluoropropene and 30 ml of p urified water were purged wi th n it rogen, sealed and mixed at 20°C fo r 1, 2 and 3 days. O n each occasion , af ter the samples had been remov ed fro m the mixer and allowed to stand for 30 minutes, an aliquot (1 ml) of each u pper aqueous phase was diluted to vol ume ( 20 ml) wi

th mobile phase fo r analysis by high performance liquid chromatography (HPLC). The pH of an undiluted portion of each sample was measured. 3.6.4 HPLC conditions Instrument: Ag ilent 1200 Series Liquid Chromatogr aph Col umn: Zorbax Eclipse XDB - C8 (15 cm x 4.6 mm int ernal diameter) Col umn tem perature: 25 oC Mobile phase composit io n: Methano l:water (3:1 v/v) Flow rate: 1.0 ml/minute Inject io n vo lu me: 1 0 μl Detector: UV set at 2 1 0 nm Retent io n t ime: Approximately 3.0 minutesHuntingdon Life Sciences WA G0005

25 3.6.5 Preparation of calibration A stock calibrat io n so lut io n of approximate concentration 1 g/l was prepared by weighing test substance (approximately 25 mg) int o a 25 ml v olumetric flask and disso lving in and dilut ing to vol ume wi th met hano l . Calibrat io n so lut io ns in the range 5 to 10 0 mg/ l were prepared by dilut io ns of t he stock sol ut io n wi th mobile phase. 3.6.6 Calculations The concentration of 2 - Bromo - 3,3,3 - trifluoropropene in the analysed so lut io n (C A) was calculated fro m standards int roduced before and af ter samples (bracket ing standards) by the fo llo wing equat io n:

C A (m g/l) = sample peak area x s tandard co ncentration (mg l)

mean peak area of br acketing s tandards The concentration of 2 - Bromo - 3,3,3 - trifluoropropene in the test sol ut io ns (C B ) was then calculated thus: C B (g/l) = ( C A (m g/l) x dilution factor)/1000 wh ere the dilut io n factor was 2 0 fo r all samples. 3.6.7 Results and discussion The detector calibrat io n was fo und to be linear over the range 0 to 1 0 0 mg/ l of s tandard sol ut io ns in mobile phase with a regressio n coefficient of 0.99 9 9 ( Table 3, Figure 3 ). Table 4 presents a summary of t he resul ts of t he test and shows that the water sol ubili ty o f 2 - Bromo - 3,3,3 - trifluoropropene was 1.01 ± 0.08 g/l . Table 5 presents the primary d ata fo r thi s test. There were no significant peaks in the chro mat ograms of blank control samples. 3.6.8 Conclusion 2 - Bromo - 3,3,3 - trifluoropropene was found to have a water solubilit y of 1.0 g/l at 20  C.Huntingdon Life Sciences WA G0005

26 Table 3 Standard calibration for 2 - Bromo - 3,3,3 -trifluoropropene by HPLC

Standard concentration (mg/l) Peak area

99.14 691.32 79.31 554.36 59.48 414.39 39.66 265.24 19.83 132.63 9.914 61.616 4.957 32.474 Linear regression: y = 7.02 x - 4.99 (including x = 0, y = 0) r = 0.99 9 9 x = concentration y = peak area Figure 3 Standard calibration for 2 - Bromo - 3,3,3 -trifluoropropene by HPLC

0 100 200 300 400 500 600 700 800

0 10 20 30 40 50 60 70 80 90 100Peak area

Concentration (mg/l)Huntingdon Life Sciences WA G0005

27 Table 4 Measurement of water solubility at 20  C Sample Time (days) Concentration Mean number at 2 0°C (g/l) concentration pH (g/l) 1A, 1B 1 0.97, 1.10 1.04 6.6, 6.5 2A, 2B 2 0.88, 0.98 0.93 7.1, 6.8 3A, 3B 3 1.04, 1.07 1.05 7.4, 7.2 Water solubility * = 1.01 ± 0.08 g/l (C. of V. = 8 %) *Determined as the mean of all samples28Huntingdon Life SciencesWAG0005

Table 5 HLPC analysis of samples from the water solubility test for 2 - Bromo - 3,3,3 -trifluoropropene

Sample Peak area C A (mg/l) Dilution factor C B ( g/l) 39.66 mg/l std 256.90 - - - Sample 3A 341.44 51.90 20 1.04 Sample 3B 352.25 53.54 20 1.07 Sample 2A 288.04 43.78 20 0.876 Sample 2B 323.13 49.12 20 0.982 39.66 mg/l std 264.92 - - - Sample 1A 317.02 48.47 20 0969 Sample 1B 360.37 55.09 20 1.10 39.66 mg/l std 253.92 - - -Huntingdon Life Sciences WA G0 005

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### 3.7 Partition coefficient (E C Method A . 8, OECD Method 117) 3.7.1 Definition and units The parti tion coefficient (P) is defined as the ratio of t he equilibrium concentrations of a di ssolved substance in a two - phase system consisting of t wo largely immiscible so lvents, in thi s case, n - octano l and water. P ow = C C octanol water The parti tion coefficient is usually given in the form of its logarithm to base ten (log 10 P ow). 3.7.2 Preliminary estimation A value of

log 10 P ow fo r 2 - Bromo - 3,3,3 - trifluoropropene was est imated by a calculat io n method using a co mputer program (KOWWIN, Version 1.67, U.S. Envi ronmental

Protection Agency, 2000). The program e st imates the lo g 10 P ow value by re ference to com pound structures for which reliable log 10 P ow data i s available. An est imated log 10 P ow value of 2.5 was calculated for 2 - Bromo - 3,3,3 - trifluoropropene. 3.7.3 Method Parti tion coefficients in the range of log 10 P ow between 0 and 6 can be est imated using high performance liquid chromatography ( HPLC). T his is perf ormed on co lu mns containing hydrocarbon chains (e.g. C 8 , C 18 ) chemically bound on to silica. T he chemicals injected on to the col umn mo ve along itb y parti tioning between the mo bile so lvent phas e and the hydrocarbon stationary phase, their retent io n being proportional to thei r hydrocarbon - water parti tion coefficient. T he parti tion coefficient is deduced fro m the capaci ty f actor, k, given by k = t - t t r o o where t r is the measured retention time of samp le or reference and t o is the retention time o f the internal deadt ime standard. 3.7.4 Procedure In order to co rrelate the measured capacit y factor (k) of t he test mat eri al wi

th i ts P ow , six reference materials were selected fro m a list of r ecommended co mpounds. These ref erence com pounds were selected to have lo g 10 P ow values between 1.0 and 6.2. A n int ernal standard (formamide) wi

th no retention on the HPLC co lu mn was used to determine the deadt ime of the HPLC system.Huntingdon Life Sciences WA G0 005

30 The fo llo wing reference mixture was prepare d in HPLC mobile phase.

Reference substance Known log 10 P ow * Concentration (mg/l) Internal deadtime standard (I.D.S.) Deadtime only 200 Acetanilide 1.0 9.84 Nitrobenzene 1.9 9.98 Ethy lbenzoate 2.6 20.6 Br omobenzene 3.0 11.9 Benzylbenzoate 4.0 11.1 DDT 6.2 21.5 *see EEC Method A8 for these values. A sol ut io n of 2 - Bromo - 3,3,3 - trifluoropropene (83.7 mg/ l in HPLC mo bile phase, and also containing I.D.S. att he same concentration as in the ref erence mixture) was prepared and chro mat ographed in d

uplicate, bracketed by samples of t he reference mixture, using the fo llo wing condit io ns. 3.7.5 HPLC conditions Instrument: Ag ilent 1200 Series Liquid Chromatograph Col umn: Zorbax Eclipse XDB - C8 (5 cm x 4.6 mm

int ernal di ameter) Col umn tem perature: 25  C Mobile phase composit io n: Methano l :water (3:1 v/v) Flow rate: 1 .0 ml/minute Inject io n vo lu me: 10 μl Detector: UV set at 210 nm 3.7.6 Results A calibrat io n plot of

log 10 k versus lo g 10 P ow was constructed fo r the ref erence standards ( Table 6, Figure 4 ). The lo garithm of t he capacit y factor fo r the test substance was then calculated and the log 10 P ow value was determined by int erpolat io n. 2 - Bromo - 3,3,3 - trifluoropropene was th us determined to have a lo g 10 P o w value of 2.7 (equivalent P o w value of 550 ). Figures 5 and 6 present examples of chromatograms generated during thi s test. 3.7.7 Conclusion 2 - Bromo - 3,3,3 - trifluoropropene was found to have a log 10 P ow val ue of 2.7 .Huntingdon Life SciencesWAG000531 Table 6 HPLC determination of log 10 P o w for 2 - Bromo - 3,3,3 - trifluoropropene Sample C omponent t r (minutes) k log 10 k log 10 P o w Standard Run A Acetanilide 0.655 0.204 - 0.690 1.0 (t 0 = 0.544 minutes) Nitrobenzene 0.799 0.469 - 0.329 1.9 Ethy lbenzoate 1.040 0.912 - 0.040 2.6 Br omobenzene 1.225 1.252 0.098 3.0 Benzylbenzoate 1.544 1 .838 0.264 4.0 DDT 6.567 11.072 1.044 6.2 Sample A (t 0 = 0.544 minutes) 2 - Br omo - 3,3,3 - trifluoropropene 1.011 0.858 - 0.066 2.7 Sample B (t 0 = 0.544 minutes) 2 - Br omo - 3,3,3 - trifluoropropene 1.010 0.857 - 0.067 2.7 Standard Run B Acetanilide 0.657 0.203 - 0.692 1.0 (t 0 = 0.546 minutes) Nitrobenzene 0.801 0.467 - 0.331 1.9 Ethy lbenzoate 1.043 0.910 - 0.041 2.6 Br omobenzene 1.227 1.247 0.096 3.0 Benzylbenzoate 1.546 1.832 0.263 4.0 DDT 6.569 11.031 1.043 6.2 Linear regressi on (based on mean of standard runs A and B) : log 10 P o w = 3.07 log 10 k + 2. 94Huntingdon Life SciencesWAG000532 Figure 4 log 10 k versus log 10 P o w calibration of standard runs A and B

0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0

- 1.000 - 0.500 0.000 0.500 1.000 1.500log10Pow

log 10 kHuntingdon Life SciencesWAG000533 Figure 5 Chromatogram of standard materials with log 10 P ow between 1.0 and 6.2

min 0 1 2 3 4 5 6 7 mAU

0 20 40 60 80 100 VW D1 A, W avelength=210 nm (0PC_00F\002-0201.D)0.5440.6550.7991.0401.2251.5446.567Huntingdon Life SciencesWAG000534 Figure 6 Chromatogram of 2 - Bromo - 3,3,3 - trifluoropropene

min 0 1 2 3 4 5 6 7 mAU

0 20 40 60 80 100 120 140 VW D1 A, W avelength=210 nm (0PC_00F\003-0301.D)0.5441.011Huntingdon Life Sciences WA G0005

35 

### 3.8 Flash point (E C Method A . 9) 3.8.1 Method The flash po intof 2 - Bromo - 3,3,3 - trifluoropropene was determined in duplicate using Pensky Martens closed cup flash po intapparatus. 3.8.2 Definition and units The flash point is d e fined as the lo west tem perature (°C) at which liquid in a closed test vessel evo lves vapour, under the condit io ns defined in the test method, in such amounts that a flammable vapour/air mixture is produced in the test vessel. 3.8.3 Apparatus Pensky - Martens Close d Cup Flash Point apparatus. Calibrat io n: The apparatus and thermo meters used have been regularly calibrated by m eans of r ef erence standards covering the appropri ate tem perature range and fo und to be wi thin acceptable limit s. 3.8.4 Procedure The test cup was fil led wi th the prescribed vo lu me of test substance. The apparatus was the n assembled in a fume cupboard. The test flame was lit and adj usted such that itw as approximately 4 mm in diameter. The test substance was st irred cont inuously except during the appli cation of

t he test flame. The test substance was heated at 5 - 6°C/minute fro m approximately

23°C , the test flame applied at 1°C int ervals and observat io ns recorded. The test was repeated using a fresh sample. The baro met ric pressure was recorded. 3.8.5 Calculatio n Pressure correction to flash points: Corrected flash point = D + 0.025 (1013 - P) where D = determined flash point (°C) P = ambient barometric pressure (mbar) and 1013 = standard pressure (mbar)Huntingdon Life Sciences WA G0005

36 3.8.6 Results The baro met ric pressure at the time of testing was 10 31 mbar . In t he duplicate determin at io ns, no flash was observed before the test substance appeared to bo il at 34°C . This observat io n precluded obtaining a flash po intvalue. 3.8.7 Conclusion No flash point was determined fo r 2 - Bromo - 3,3,3 - trifluoroprope ne before the sample bo iled. The starting tem

perature fo r thi s test was approximately 23°C, but thi s resul t covers all tem peratures below the bo iling point. I f flammabilit y was observed during this test, addi tional work at colder tem

peratures would have b een conducted.Huntingdon Life Sciences WA G0005

37 

### 3.9 Explosive properties (E C Method A . 14) It was considered that the test substance was not explosive and therefore an assessment of thi s property

was made using a theoretical consideration of its structure. 3.9.1 Assessment of explosive propertie s Chemical regulatory

authori ti es requi re information on the explosive properti es of substances fo r classificat io n and labelling purposes. Test met hods fo r the determinat io n of e xplosive properti es (EEC Method A14) invo lve measuring the effect of t hree stimu li; shock, heat and fr ict io n. The test substance is considered to p ossess explosive properti es if a posi tive resul t is obtained in any one of these tests. According to the United Nat io ns Co mmittee of Experts on the Transport of Dangerous Goods explo sive properti es are associ ated wi th t he presence of

c ertain chemical groups in a mo lecule which can react to produce a very r apid increase in tem

perature or pressure. T hus, iti s possible to examine the chemical structure of a substance and ascertain whether o r not there are any groups present that are associ ated with explosive properti es. For this purpose a list of screening procedures has been published in Appendix 6 of the Uni ted Nat io ns ‘Reco mmendations on the Transport of Dangerous Goods, Manual o f Tests and Criteria’. The init ia l assessment of e xplo sive properti es is made by examining the chemical structures of t he com ponents wi

th a l ist of f unct io nal groups that ma y indicate explosive properti es in organic materials. This list i s reproduced below: Struct ural feature Examples C - C unsaturation Acetylenes, acetylides, 1,2 - dienes C - Metal, N - Metal Grignard reagents, organo - lithium compounds Contiguous nitrogen atoms Azides, aliphatic azo compounds, diazonium salts, hy drazines, sulphonylhydrazides Contiguou s oxy

gen atoms Peroxides, ozonides N - O Hydroxylamines, nitrates, nitro compounds, nitroso compounds, N - oxides, 1,2 - oxazoles N - halogen Chloramines, fluoroamines O - halogen Chlorates, perchlorates, iodosyl compounds While 2 - Bromo - 3,3,3 - trifluoropropene c ontains C - C unsaturation, iti s a single C - C double bond . E xplo sive properti es are associated wi th com pounds having a greater degree of unsaturation , as listed in the examples above. From a n examinat io n of

the chemical structure of t he test substance, itw as concl uded that itc ontained no fu nct io nal groups which are characterist ic o f explo sive properti es. 3.9.2 Conclusion From the assessment of the chemical structure it was established that the test substance, 2 - Bromo - 3,3,3 - trifluoropropene , would not have explosive properti es.Huntingdon Life Sciences WA G0005

38 

### 3.10 Auto - ignition temperature (liquids and gases) (E C Method A . 15) 3.10.1 Method The auto - ig nit io n tem

perature was evaluated using the method described in BS 4056. 3.10.2 Definition and units The auto - ig nit io n tem

perature is the lowest tem perature at which the test substance will ignite when mixed wi

th a iru nder the condi

tions defined in the test met hod. T he units are degrees Celsius (°C). 3.10.3 Apparatus Furnace consist ing of

a r ef ractory c ylinder, manufactured by Carbo lit e, fit ted wi th thermocouples for tem

p erature control

and measurement. 3.10.4 Procedure A known quant it y of 2 - Bromo - 3,3,3 - trifluoropropene was injected into a heated open Erlenmeyer flask containing ai r at 400°C. The contents of t he flask were observed in a darkened room for five minutes fo llo wing ad di tion of a sample or un

til igni tion occurred. Igni tion was not observed wi thin five minutes so the test was prepared wit h differing sample vo lu mes to a maximum o f 1000  l. In no case was ignit io n observed within five minutes . 3.10.5 Results At mo spheric pressure : 10 31 mbar Test temperature (°C) Test volume (μl) Time (s) Observation 400 70 >300 No ignition 400 1000 >300 No ignition 400 100 >300 No ignition 400 200 >300 No ignition 400 500 >300 No ignition 3.10.6 Conclusion 2 - Bromo - 3,3,3 - trifluoropropene was determined not to auto - ig nit e below 400°C .Huntingdon Life Sciences WA G0005

39 

### 3.11 Oxidising properties (E C Method A . 21) It was considered that the test substance was not oxidising and therefore an assessment of the property was made on a theoretical considerat io n of it s structure. 3.11.1 Assessm ent of oxidising properties Chemical regulatory

a uthori ti es requi re informat ion on the oxidising properti es of substances fo r classificat io n and labelling purposes. Test met hods fo r the determination of o xidising properti es (E C Method A . 21) invo lve measuring the potenti al f

or a material to increase the burning rate of a combust ible substance. The burning rates are then co mpared to those obtained fo r the co mbust ible material mixed wi th a r eference oxidiser. T he test substance is considered to possess oxidis ing properti es if any of the test mixtures burn faster than the reference mixture. It is possible to predict the oxidising potential of a mol ecule by analogy

w it h exist ing com pounds and therefore iti s possible to examine the chemical structure of a s ubsta nce and ascertain whether or not there are any groups present that are associated wi th o xidising properties. F or thi s purpose a screening procedure has been published in Appendix 6 of t he United Nat io ns ‘Reco mmendat io ns on the Transport of D angerous Goods , Manual of T ests and Criteria’. For organic co mpounds, the substance does not possess oxidising properti es if itd oes not contain oxygen, fluorine or chlorine. If these atoms are present, then the substance does not possess oxidising properties if the ato ms are chemically bonded only to carbon and hydrogen. From a n examinat io n of

t he chemical structure of 2 - Bromo - 3,3,3 - trifluoropropene , it was confirmed that itd idc ontain fluorine , but only chemically bonded to carbon . Consequent ly, the test substance wou ld not be expected to possess oxidising properties. 3.11.2 Conclusion From the assessment of the chemical structure it was established that the test substance, 2 - Bromo - 3,3,3 - trifluoropropene , would not have oxidising properties.Huntingdon Life Sciences WA G0005

40 

### 3.12 Soil adsorption (E C Method C . 19, OECD Method 121) 3.12.1 Definition and units The adsorption coefficient is defined as the ratio b etween the concentration of t he substance in the so il and the concentration of the substance in the aqueous phase, both measured when adsorption equilibrium is reach ed. The adsorption coefficient is usually given in the form of its logarithm to base ten (log 10 K oc ). 3.12.2 Method The sorption behaviour of the test substance on so il can be invest igated using high performance liquid chromatography ( HPLC). T his is perf

ormed on co mmercially available analyt ical col

umns packed wi

th a solid phase containing a mo derately polar stationary phase wi th l ipophilic and polar mo ieties (e.g. cyano - propyl bonded phase). The chemicals injected onto the col

umn mo ve along itb y part it io ning bet ween the mo bile phase and the stationar y phase. T he velocit y of

e ach co mponent thereby depends on the degree of a dsorption on the stationary phase. T he dual nature of t he stationary phase allows fo r int eraction of p olar and apolar parts of

a molecule in a similar way as is the case fo r soil. T his enables the relat io nship between the retention time on such a col umn and the adsorption coefficient on the organic parts of the soil to be established. The adsorption coefficient is deduced fro m the capacit y fa c tor (k) given by: k = t - t t r o o where t r is the measured retention time o f sample or reference and t o is the retention time o f the internal deadt ime standard 3.12.3 Procedure In order to co rrelate the me asured capacit y factor (k) of t he test mat e ri al wi

th i ts K oc , six reference materials were selected fro m a list of r ecommended co mpounds. T hese ref erence com pounds were selected to have lo g 10 K oc values between 1.3 and 5.6. An int ernal standard (formamide) wi

th no retention on the HPLC co lu mn was u sed to determine the deadt ime of the HPLC system.Huntingdon Life Sciences WA G0005

41 The fo llo wing reference mixture was prepared in HPLC mobile phase.

Reference substance Known log 10 K oc * Concentration (mg/l) Internal deadtime standard (I.D.S.) Deadtime only 20 0 Acetanilide 1.3 9.84 Nitrobenzene 2.4 9.98 Naphthalene 2.8 5.02 2 - Methy lnaphthalene 3.9 5. 58 Anthracene 4.4 22.8 DDT 5.6 21.5 *values from OECD Method 121, with the exception of nitrobenzene, 2 - methy lnaphthalene and anthracene, values for wh ich came from ‘HODS ON, J. and WILLIAMS, N.A., (1988). The Estimation of the Adsorption Coefficient (K

oc ) for Soils by High Performance Liquid Chromatography, Ch emosphere, Vol. 17, pp.67 - 77, Pergammo

n Journals Ltd’. A sol ut io n of 2 - Bromo - 3,3,3 - trifluoropropene ( 71.1 m g/l in HPLC mo bile phase, and also containing I.D.S. att he same concentration as in the ref erence mixture) was prepared and chro mat ographed in duplicate, bracketed by samples of t he reference mixture, using the fo llo wing condit io ns. 3.12.4 HPLC conditions Instrument: Agilent 1200 Seri es Liquid Chromatograph Col umn: Hy perClone CN (CPS) ( 2 5 cm x 4.6 mm int ernal diameter) Col umn tem perature: 25°C Mobile phase composit io n: Methano l : water (55:45 v/v) Flow rate: 1.0 ml/minute Inject io n vo lu me: 20 μl D etector: UV set at 210 nmHuntingdon Life Sciences WA G0005

42 3.12.5 Results A calibrat io n plot of

log 10 k versus lo g 10 K oc was constructed fo r the reference standards ( Table 7 , Figure 7 ). The lo gari thm o f the capacit y factor fo r the test substance was then calculated and the lo g 10 K oc value was determined by int erpolat io n. 2 - Bromo - 3,3,3 - trifluoropropene was th us determined to have a lo g 10 Ko c val ue of 2.2 (equivalent Ko c val

ue of 150 ). Figures 8 and 9 present examples of chromatograms generated during thi s test . 3.12.6 Conclusion 2 - Br omo - 3,3,3 - trifluoropropene was fo und to have a lo g 10 K oc value of 2.2 .43Huntingdon Life SciencesWAG0005

Table 7 HPLC determination of log 10 K oc for 2 - Bromo - 3,3,3 - trifluoropropene

Sample Component t r (minutes) k log 10 k log 10 K oc Standard Run A Acetanilide 3.672 0.189 - 0. 724 1.3 (t 0 = 3.089 minutes) Nitrobenzene 4.360 0.411 - 0.386 2.4 Naphthalene 5.470 0.771 - 0.113 2.8 2 - Methy lnaphthalene 6.184 1.002 0.001 3.9 Anthracene 8.508 1.754 0.244 4.4 DDT 14.670 3.749 0.574 5.6 Sample A (t 0 = 3.066 minutes) 2 - Br omo - 3,3,3 - trifluoropropene 4.228 0.379 - 0.421 2.2 Sample B (t 0 = 3.061 minutes) 2 - Br omo - 3,3,3 - trifluoropropene 4.223 0.380 - 0.421 2.2 Standard Run B Acetanilide 3.631 0.188 - 0.725 1.3 (t 0 = 3.056 minutes) Nitrobenzene 4.303 0.408 - 0.389 2.4 Naphthalene 5.383 0.761 - 0.118 2.8 2 - Methy lnaphthalene 6.080 0.990 - 0.005 3.9 Anthracene 8.360 1.736 0.239 4.4 DDT 14.568 3.767 0.576 5.6 Linear regression (based on mean of standard runs A and B) : log 10 K oc = 3.39 log 10 k + 3.6144Huntingdon Life SciencesWAG0005

Figure 7 lo g 10 k versus log 10 K oc calibration of standard runs A and B

0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0

- 1.000 - 0.800 - 0.600 - 0.400 - 0.200 0.000 0.200 0.400 0.600 0.800 1.000log10Koc

log 10k45Huntingdon Life SciencesWAG0005

Figure 8 Chromatogram of standard materials with log 10 K oc between 1.3 and 5.6

min 0 2.5 5 7.5 10 12.5 15 17.5 mAU

0 50 100 150 200 250 300 VW D1 A, W avelength=210 nm (1PC_00F\001-0101.D)3.0893.6724.3605.4706.1848.50814.67046Huntingdon Life SciencesWAG0005

Figure 9 Chromatogram of 2 - Bromo - 3,3,3 - trifluoropropene

min 0 2.5 5 7.5 10 12.5 15 17.5 mAU

0 20 40 60 80 100 120 140 VW D1 A, W avelength=210 nm (1PC_00F\002-0201.D)3.0664.228Huntingdon Life Sciences WA G0005

47 Annex 1 Eye R e search Centre GLP Compliance Statement 20 1 1