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Working for a better future

2-Bromo-3,3,3-Trifluoropropene

Abiotic Degradation: Hydrolysis as a Function of pH 

(Preliminary Test)

HLS study number: WAG0007

Version ID: Final

Issue date: 26 July 2012
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Sponsor and Test Facility 

Sponsor American Pacific Corporation

3883 Howard Hughes Parkway 

Suite 700

Las Vegas 

NV 89169

USA

Test facility Huntingdon Life Sciences

Eye Research Centre

Eye

Suffolk

IP23 7PX

UK
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Table of Contents

Sponsor and Test Facility...................................................................................................... 2

Compliance with Good Laboratory Practice .......................................................................... 4

Quality Assurance Statement................................................................................................ 5

Contributing Scientists .......................................................................................................... 6

Summary .............................................................................................................................. 7

1. Introduction .................................................................................................................. 8

1.1 Objective........................................................................................................... 8

1.2 Regulatory compliance ..................................................................................... 8

1.3 Study schedule ................................................................................................. 8

1.4 Maintenance of records .................................................................................... 8

2. Test Substance ............................................................................................................ 9

3. Abiotic Degradation: Hydrolysis as a Function of pH (Preliminary Test) .................... 10

3.1 Method ........................................................................................................... 10

3.2 Definition and units ......................................................................................... 10

3.3 Procedure....................................................................................................... 10

3.3.1 Preparation of buffer solutions............................................................. 10

3.3.2 Preliminary test ................................................................................... 10

3.4 GC conditions ................................................................................................. 11

3.5 Preparation of calibration ................................................................................ 12

3.6 Calculations.................................................................................................... 12

3.7 Results ........................................................................................................... 12

3.7.1 Linearity .............................................................................................. 12

3.7.2 Method verification .............................................................................. 12

3.7.3 Preliminary test ................................................................................... 12

3.7.4 pH of test solutions and incubation temperature.................................. 13

3.8 Conclusion...................................................................................................... 13

List of Annexes

Annex 1 Eye Research Centre GLP Compliance Statement 2011 .............................23


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Contributing Scientists

2-Bromo-3,3,3-Trifluoropropene: 

Abiotic degradation: hydrolysis as a function of pH (preliminary test)

Study management

P Sydney BSc MSc

Study Director 

Product Chemistry

Other responsible staff

L Lawson MChem

Scientist

Product Chemistry
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Summary

A study was performed to determine the rate of hydrolysis of 2-Bromo-3,3,3-

Trifluoropropene as a function of pH. The test was conducted in accordance with the 

preliminary test as described in Commission Regulation (EC) No. 440/2008 (Method C.7) 

and the OECD Guidelines for Testing of Chemicals (Method 111).

The preliminary study showed that at each of pH 4, 7 and 9 and 50  0.5oC, less than 10%

hydrolysis had occurred after 5 days, equivalent to a half-life of greater than 1 year under 

environmental conditions (25°C). No further testing was considered necessary.

2-Bromo-3,3,3-Trifluoropropene was determined to be hydrolytically stable under acidic,

neutral and basic conditions.
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1. Introduction

1.1 Objective

This study was designed to determine the rate of hydrolysis of 2-Bromo-3,3,3-

Trifluoropropene as a function of pH. Determination of the rate of hydrolysis of a test 

substance aids in predicting its environmental fate. 

1.2 Regulatory compliance

The test was conducted in accordance with the preliminary test as described in Commission

Regulation (EC) No. 440/2008 (Method C.7) and the OECD Guidelines for Testing of 

Chemicals (Method 111).

1.3 Study schedule 

Study initiation date : 06 December 2011

Experimental start date : 20 January 2012

Experimental completion date : 12 March 2012

Study completion date : 26 July 2012

1.4 Maintenance of records

Primary data from the test, and a copy of the final report, are stored in the archives of 

Huntingdon Life Sciences.
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2. Test Substance

Identity: 2-Bromo-3,3,3-trifluoropropene

Chemical structure: F

F

F

CH2

Br

CAS number: 1514-82-5

Appearance: Clear liquid with a slight yellow tint

Storage conditions: Room temperature

Lot number: KgF080

Purity: 99.9% excluding stabilisers

Stabilisers:

Expiry date: 01 August 2012

Date received: 12 September 2011
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3. Abiotic Degradation: 

Hydrolysis as a Function of pH (Preliminary Test) 

(EC Method C.7, OECD Method 111)

3.1 Method

The hydrolysis of the test substance was investigated according to the preliminary test as 

described in EC Method C.7 and OECD Method 111.

3.2 Definition and units

Hydrolysis refers to a reaction of a chemical RX with water. This reaction may be 

represented by the net exchange of the X with OH:

RX + HOH → ROH + HX

The concentrations can be expressed in units such as milligrams per litre or moles per litre.

3.3 Procedure

Tests were carried out using reagent grade chemicals, purified sterile water and sterilised 

glassware.

3.3.1 Preparation of buffer solutions

Buffer solutions were prepared as follows:

pH 4: 0.2M aqueous potassium dihydrogen orthophosphate (55 ml) was mixed with

0.07M aqueous disodium hydrogen orthophosphate dodecahydrate (125 ml) and purified 

water (320 ml). The pH was adjusted to 4.0  0.05 with orthophosphoric acid.

pH 7: 0.2M aqueous potassium dihydrogen orthophosphate (125 ml) was mixed with

1M sodium hydroxide (15 ml) and purified water (360 ml). The pH was adjusted to 7.0 

0.05 with 1M hydrochloric acid.

pH 9: 0.1M boric acid in 0.1M aqueous potassium chloride (250 ml) was mixed with

1M sodium hydroxide (10.5 ml) and purified water (240 ml). The pH was adjusted to 9.0 

0.05 with 1M hydrochloric acid.

3.3.2 Preliminary test

The following procedure was carried out at each of pH 4, 7 and 9:

Aliquots (100 μl) of a stock solution of 2-Bromo-3,3,3-Trifluoropropene in methanol

(approximately 2.5 g/l) were added to separate 10 ml Wheaton vials containing buffer 

solution (9.9 ml), which had been purged with nitrogen and pre-equilibrated at test 

temperature (50 ± 0.5°C).
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The samples, of nominal concentration 25 mg/l, were placed in a 50°C waterbath in the dark 

until sampling was required (after a short incubation period of approximately 5 minutes, and 

then after 2.4, 48, 120 and 240 hours).

At each sampling time, two samples were removed from the bath. After cooling, each vial 

was opened and the entire volume was quantitatively transferred to a separate vessel with a

portion (10 ml) of hexane. After shaking for 90 seconds, the upper hexane phase was 

analysed by gas chromatography (GC). 

Sample pH and incubation temperature were monitored over the period of the test.

Method verification samples were prepared at room temperature at each pH value, the 

samples being fortified and processed as above.

3.4 GC conditions

Instrument: Agilent 5890 Series Gas Chromatograph

Column: DB-5 (30 m x 0.32 mm internal diameter x 1.00 m film thickness)

Carrier gas: Helium

Flow: 2.0 ml/minute at 35°C

Oven temperature

Initial: 35°C (5 minutes)

Ramp: 5°C/minute

Final: 80°C (2 minutes)

Injection temperature: 175°C

Injection technique: Split (1:25)

Injection volume: 1 μl

Detector: ECD

Detector gas flows

Anode purge (N2): 10 ml/minute

Make-up (N2) 80 ml/minute

Detector temperature: 320C

Retention time: Approximately 2.7 minutes
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3.5 Preparation of calibration

A stock calibration solution of approximate concentration 500 mg/l was prepared by adding 

test substance (30 μl; density = 1.653 g/ml, Huntingdon Life Sciences Study Number

WAG0005) to a 100 ml volumetric flask containing hexane and dissolving in and diluting to 

volume with the same.

Calibration solutions in the range 5 to 50 mg/l were prepared by dilutions of the stock 

solution with hexane.

3.6 Calculations

The concentration of 2-Bromo-3,3,3-Trifluoropropene in the analysed solution (CA) was 

calculated from standards introduced before and after samples (bracketing standards) by the 

following equation: 

CA (mg/l) = 

mean peak area of bracketing standards

sample peak area x standard concentration (mg l)

3.7 Results

3.7.1 Linearity

Under the conditions described, the calibration of 2-Bromo-3,3,3-Trifluoropropene was found 

to be linear over the concentration range 0 to 50 mg/l, with a regression coefficient of 0.9992

(Table 1, Figure 1). An example standard chromatogram is shown in Figure 2.

It was considered that the analytical method was sufficiently sensitive to quantify test 

substance concentrations down to 10% or less of the initial concentration.

3.7.2 Method verification

An overall mean recovery value of 91% was determined, verifying the efficiency of sample 

processing.

3.7.3 Preliminary test

The preliminary study showed that at each of pH 4, 7 and 9 and 50  0.5oC, less than 10% 

hydrolysis had occurred after 120 hours (5 days), equivalent to a half-life of greater than 

1 year under environmental conditions (25°C). No further testing was considered necessary.

Table 2 summarises the results from the preliminary test, whilst Table 3 presents the primary 

data. Example chromatograms are shown in Figures 3 to 8. Although slow hydrolysis was 

evident, the t240h chromatograms did not show additional degradate peaks. It is possible that 

any degradation products may have been more water soluble than the parent compound and 

did not partition into the extraction solvent (hexane).
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3.7.4 pH of test solutions and incubation temperature

The pH values of test solutions and incubation temperature over the period of the test are 

presented in Table 4. The results show that there were no significant changes in pH and 

temperature with time.

3.8 Conclusion

2-Bromo-3,3,3-Trifluoropropene was determined to be hydrolytically stable under acidic, 

neutral and basic conditions.
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Table 1 Standard calibration for 2-Bromo-3,3,3-Trifluoropropene

Standard concentration (mg/l) Peak area

49.54 2270800

39.63 1833100

29.72 1392800

19.82 991250

9.908 536170

4.954 291960

Linear regression: y = 44990x + 58040 

(including x=0, y=0) r = 0.9992

x = concentration

y = peak area
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Table 2 Preliminary test results for hydrolysis of 2-Bromo-3,3,3-Trifluoropropene

Ct (mg/l)

pH t0h t2.4h t48h t120h t240h

Measured Mean Measured Mean Measured Mean Measured Mean Measured Mean

4 22.9, 21.8 22.3 20.1, 21.3 20.7 20.2, 20.5 20.3 19.8, 18.2 19.0 18.0, 18.3 18.1

7 21.1, 22.0 21.6 19.7, 19.8 19.7 20.3, 18.1 19.2 17.8, 19.9 18.9 18.1, 18.2 18.1

9 22.9, 22.0 22.5 21.5, 21.4 21.4 19.9, 19.2 19.5 20.2, 20.0 20.1 17.8, 19.7 18.7

where Ctis the concentration of 2-Bromo-3,3,3-Trifluoropropene in solution at time th (in hours)

At pH 4, ln Ct = 3.05 - 0.000714t; t0 = 21.2 mg/l, t120 = 19.5 mg/l; Hydrolysis at t120 = 8%

At pH 7, ln Ct = 3.01 - 0.000505t; t0 = 20.3 mg/l, t120 = 19.1 mg/l; Hydrolysis at t120 = 6%

At pH 9, ln Ct = 3.06 - 0.000587t; t0 = 21.4 mg/l, t120 = 19.9 mg/l; Hydrolysis at t120 = 7%
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Table 3 Measurement of 2-Bromo-3,3,3-Trifluoropropene in hydrolysis 

test solutions

Sample Peak area CA (mg/l)

19.82 mg/l std 959030 -

pH 4/t0 A 1125100 22.89

pH 4/t0 B 1069100 21.75

pH 7/t0 A 1037200 21.10

pH 7/t0 B 1083100 22.03

19.82 mg/l std 989660 -

pH 9/t0 A 1139100 22.94

pH 9/t0 B 1094000 22.03

pH 4/t2.4 A 998570 20.11

pH 4/t2.4 B 1056100 21.27

19.82 mg/l std 978910 -

pH 7/t2.4 A 981540 19.67

pH 7/t2.4 B 986880 19.78

pH 9/t2.4 A 1073000 21.51

pH 9/t2.4 B 1066200 21.37

19.82 mg/l std 998910 -

pH 4/t48 A 1011300 20.15

pH 4/t48 B 1026100 20.45

pH 7/t48 A 1016500 20.26

pH 7/t48 B 908840 18.11

19.82 mg/l std 990150 -

pH 9/t48 A 1015700 19.87

pH 9/t48 B 981250 19.20

pH 4/t120 A 1010500 19.77

pH 4/t120 B 928340 18.16

19.82 mg/l std 1036200 -

pH 7/t120 A 951140 17.84

pH 7/t120 B 1059300 19.86

pH 9/t120 A 1076300 20.18

pH 9/t120 B 1065900 19.99

19.82 mg/l std 1077700 -

19.82 mg/l std 975780 -

pH 4/t240 A 893770 17.95

pH 4/t240 B 910070 18.28

pH 7/t240 A 899690 18.07

pH 7/t240 B 906750 18.21

19.82 mg/l std 997760 -

pH 9/t240 A 892030 17.76

pH 9/t240 B 989290 19.70

19.82 mg/l std 993090 -
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Table 4 pH change and temperature during preliminary hydrolysis tests

Nominal pH Initial pH Final pH Initial temperature 

(°C)

Final temperature 

(°C)

4 4.0 4.1 50 50

7 7.1 7.1 50 50

9 9.0 9.0 50 50
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Figure 1 Standard calibration for 2-Bromo-3,3,3-Trifluoropropene

0.0E+00

5.0E+05

1.0E+06

1.5E+06

2.0E+06

2.5E+06

0 5 10 15 20 25 30 35 40 45 50

Peak area

Concentration (mg/l)
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Figure 2 Example standard chromatogram (19.82 mg/l)

0 2 4 6 8 10 12 14 min

counts

0

100000

200000

300000

400000

 ECD1 B, (0HY_00F\010F1001.D) 2.693
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Figure 3 Example pH 4 t0h chromatogram

0 2 4 6 8 10 12 14 min

counts

0

100000

200000

300000

400000

500000

 ECD1 B, (0HY_00F\011F1101.D) 2.692

Figure 4 Example pH 4 t240h chromatogram

0 2 4 6 8 10 12 14 min

counts

0

100000

200000

300000

400000

 ECD1 B, (3HY_00F\011F1101.D) 2.706
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Figure 5 Example pH 7 t0h chromatogram

0 2 4 6 8 10 12 14 min

counts

0

100000

200000

300000

400000

500000

 ECD1 B, (0HY_00F\013F1301.D) 2.693

Figure 6 Example pH 7 t240h chromatogram

0 2 4 6 8 10 12 14 min

counts

0

100000

200000

300000

400000

 ECD1 B, (3HY_00F\013F1301.D) 2.705
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Figure 7 Example pH 9 t0h chromatogram

0 2 4 6 8 10 12 14 min

counts

0

100000

200000

300000

400000

500000

 ECD1 B, (0HY_00F\016F1601.D) 2.693

Figure 8 Example pH 9 t240h chromatogram

0 2 4 6 8 10 12 14 min

counts

0

50000

100000

150000

200000

250000

300000

350000

400000

450000

 ECD1 B, (3HY_00F\016F1601.D) 2.705
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Annex 1 Eye Research Centre GLP Compliance Statement 2011