## FINAL REPORT

Study Title



## ASSESSMENT OF ACUTE ORAL TOXICITY WITH

1- PROPANESULFONIC ACID , 2

- HYDROXY -3-(2-PROPEN-1-YLOXY)-

,

SODIUM SALT (1:1)

IN THE RAT (UP AND DOWN PROCEDURE)

Author

Drs. A.H.B.M. van Huygevoort

Test Facility

WIL Research Europe B. V.

Hambakenwetering 7

5231 DD ‘s

- Hertogenbosch

The Netherlands

Laboratory Project Identification

Project 500679

Substance 204240/A1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 2 - 1. CONTENTS

1. CONTENTS

................................................................................................ ...................................... 2

2. STATEMENT OF GLP COM PLIANCE

............................................................................................. 3

3. QUALITY ASSURANCE ST ATEMENT

............................................................................................ 4

4. SUMMARY

................................................................................................ ........................................ 5

5. INTRODUCTION

.............................................................................................................................. 6

5.1. Preface

................................................................................................ ..................................... 6

5.2. Aims of the study

...................................................................................................................... 6

5.3. Guidelines

................................................................................................ ................................ 6

5.4. Storage and retention of records and materials

....................................................................... 7

6. MATERIALS AND METHOD S

.......................................................................................................... 8

6.1. Test substance

......................................................................................................................... 8

6.1.1. Test substance information

............................................................................................. 8

6.1.2. Study specific test substance information

...................................................................... 8

6.1.3. Test substance preparation

............................................................................................ 8

6.2. Test system

.............................................................................................................................. 8

6.3. Animal husbandry

.................................................................................................................... 9

6.4. Study design

............................................................................................................................ 9

6.5. Treatment

................................................................................................ ................................ . 9

6.6. Observations

.......................................................................................................................... 10

6.7. Electronic data capture

.......................................................................................................... 10

6.8. Interpretation

.......................................................................................................................... 10

6.9. List of deviations

.................................................................................................................... 10

6.9.1. List of protocol deviations

............................................................................................. 10

6.9.2. List of standard operating procedures deviations

......................................................... 10

7. RESULTS

................................................................................................ ....................................... 11

7.1. Mortality (Table 1)

.................................................................................................................. 11

7.2. Clinical signs (Table 2)

........................................................................................................... 11

7.3. Body weights (Table 3)

.......................................................................................................... 11

7.4. Macroscopic findings (Table 4)

.............................................................................................. 11

8. CONCLUSION

................................................................................................ ................................ 11

T ABLES

Table 1: Mortality

................................................................................................ .................................... 12

Table 2: Clinical s igns

............................................................................................................................. 12

Table 3: Body w eights

............................................................................................................................ 12

Table 4: Macroscopic f indings

................................................................................................................ 13

APPE NDICES

APPENDIX 1: TEST SUBSTANCE CERTIFICATE OF ANALY SIS

....................................................... 141- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 5 - 4. SUMMARY

Assessment of acute oral t oxicity with 1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) in the rat (Up and Down procedure).

The study was carried out based on the guidelines described in:

OECD No.425 (2001) "Acute Oral Toxicity

– Up and Down procedure",

EPA, OPPTS 870.1100 (2002), "Acute Oral Toxicity"

1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) was administered by oral

gavage to five female Wistar rats at 2000 mg/kg body weight. Animals were subjected to daily

observatio ns and weekly determination of body weight. Macroscopic examination was performed a fter

terminal sacrifice (Day 15).

No mortality occurred.

Hunched posture and/or piloerection were seen in two females

The body weight gain shown by the animals over the study period was considered to be similar to that

expected of normal untreated animals of the same age and strain

.

No abnormalities were found at macroscopic post mortem examination of the animals.

The oral LD50 value of 1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) in

Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results , 1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1)

does not have to be classified and has no obliga tory labelling requirement for acute oral toxicity

according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of

the United Nations ( 2011 ) and Regulation (EC) No 1272/2008 on classification, labelling and

packaging of substances and mixtures.1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 6 - 5. INTRODUCTION

5.1. Preface

Sponsor Society of Chemical Manufacturers and Affiliates

1850 M Street, NW Suite 700

WASHINGTON, DC 20036

USA

Study Monitor Dr. C. T. Helmes

Test Facility WIL Research Europe B.V.

Hambakenwetering 7

523 1 DD 's

- Hertogenbosch

The Netherlands

Study Director Drs. A.H.B.M. van Huygevoort

Study Plan (in

- life phase) Start : 17 January 2013

Completion : 07 February 2013

5.2. Aims of the study

The objective of this study was to assess the toxicity of the test substance when administered in a

single dose to female rats at one or more defined dosages. Furthermore, the results of the study

allowed the test substance to be ranked according to most classification systems, currently in use.

This study should provide a rational basis for risk assessment in man.

The oral route was selected as it is a possible route of human exposure during manufacture, handling

or use of the test substance.

5.3. Guidelines

As required by the Dutch Act on Animal Experimentation (February 1997), this type of protocol was

reviewed and agreed by the Laboratory Animal Welfare Officer and the Ethical Committee (DEC 05

- 15) as required by the Dutch Act on Animal Experimentation (February 1997). The study procedures

described in this protocol are based on the following guidelines:

Organization for Economic Co

- operation and Development (OECD), OECD Guidelines for Testing of

Chemicals, Section 4, Health Effects, No.425, "Acute Oral Toxicity

– Up and Down procedure", Paris

Cedex, 200 8.

United Stat es Environmental Protection Agency (EPA). Health Effects Test Guidelines, OPPTS

870.1100, Acute Oral Toxicity. Office of Prevention, Pesticides and Toxic Substances (7101), EPA

712 -C- 02

- 190, 2002.1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 7 - 5.4. Storage and retention of records and materials

Records and materials pertaining to the study including protocol, raw data and the final report are

retained in the WIL Research Europe archives for a period of at least 2 years after finalization of the

report. After this period, the sponsor will be contacted to determine how the records and materials

should be handled. WIL Research Europe will retain information concerning decisions made.

WIL Research Europe will retain a test substance sample until the expiry date, but no longer than 10

years after finalization of the report. After this period the sample will be destroyed.1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 8 - 6. MATERIALS AND METHOD S

6.1. Test substance

6.1.1. Test substance information

Identification 1

- Propanesulfonic acid, 2

- hydroxy

- 3

- (2

- propen

- 1

- yloxy)

- , sodium salt (1:1)

Synonyms COPS_1 or CS

- AHPS

Structu re

O SO 3 -

OH Na +

Molecular formula C 6 H 11 NaO 5 S

Molecular weight 218.22

Description Clear colourless liquid

Batch 12E1101

Purity/Composition

Sodium -1- allyloxy -2-

hydroxypropane sulphonate

3- allyl

- 1,2

- propanediol

Water CAS Number Percent

52556

- 42

- 0 38.64

123

- 34

- 2 2.26

7732

- 18

- 5 59.1

Test substance storage At room temperature in the dark

Stability under storage conditions Stable

Expiry date 13 August 2013

See appendix I; Certificate of analysis.

6.1.2. Study specific test substance information

Purity/composition correction factor

required No

Hygroscopic No

Volatile No

Specific Gravity / Density 1.17

pH 7 at concentration of 40.9%

Stability at higher temperatures Yes, maximum temperature: 80 ° C,

maximum duration: 1 hour

6.1.3. Test substance preparation

The test substance was dosed undiluted as delivered by the sponsor.

No correction was made for the purity of the test substance.

6.2. Test s ystem

Species Rat, Wistar strain Crl:WI (outbred, SPF

- Quality). Recognized by

international guidelines as the recommended test system (e.g. OECD,

EC).

Source: Charles River Deutschland, Sulzfeld, Germany.

Number of animals 5 Females (nulliparous and non

- pregnant).

Age and body weight Young adult animals (approx. 9-10 weeks old) were selected. Body

weight variation did not exceed +/

- 20% of the sex mean.

Identification Earmark and tail mark

. Health inspection At least upon receipt of the animals and prior to dosing.1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 9 - 6.3. Animal husbandry

Conditions

Environmental controls for the animal room were set to maintain 18 to 24°C, a relative humidity of 40

to 70%, approximately 15 room air changes/hour, and a 12

- hour light/12

- hour dark cycle. Any

variations to these conditions were maintained in the raw data and had no effect on the outcome of the

study.

Accommodation

Individually housed in labeled Ma k rolon cages (MIII type, height 18 cm.) containing sterilized sawdust

as bedding material (Litalabo, S.P.P.S., Argenteuil, France) and paper as cage

- enrichment (Enviro

- dri,

Wm. Lill ico & Son (Wonham Mill Ltd), Surrey, United Kingdom).

Acclimatization period

The acclimatization period was at least 5 days before the start of treatment under laboratory

conditions. Accommodation was as described above except that the animals were group housed in

labeled Ma k rolon cages (MIV type; height 18 cm).

Diet

Free access to pelleted rodent diet (SM R/M

- Z from SSNIFF® Spezialdiäten GmbH, Soest, Germany).

Water

Free access to tap water.

Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed

according to facility standard procedures. There were no findings that could interfere with the study.

6.4. Study design

The toxicity of the test substance was assessed by stepwise treatment of five females. The first animal

was treated at a dose level of 2000 mg/kg. The absence or presence of mortality at one step

determined the next step, based on the test procedure defined in the guidelines. Based on the results

of this first animal, four additional animals were dosed at 2000 mg/kg.

6.5. Treatment

Method Oral gavage, using plastic feeding tubes

.

Fasting Animals were deprived of food overnight prior to dosing and until 3

- 4

hours after administration of the test substance. Water was available.

Frequency Single dosage, on Day 1.

Dose level (volume) 2000 mg/kg ( 1.71 mL /kg) body weight.

Dose volume calculated as dose level (g/kg) / specific gravity.1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 10

- 6.6. Observations

Mortality/Viability Twice daily.

Body weights Days 1 (pre

- administration), 8 and 15

.

Clinical signs At period ic intervals on the day of dosing (Day 1) and once daily

thereafter, until Day 15. The signs were graded according to fixed scales

and the time of onset, degree and duration were recorded:

Maximum grade 4: grading slight (1) to very severe (4)

Maximum grade 3: grading slight (1) to severe (3)

Maximum grade 1: presence is scored (1).

Necropsy At the end of the observation period, all animals were sacrificed by

oxygen/carbon dioxide procedure and subjected to necropsy.

Descriptions of all internal macroscopic abnormalities were recorded.

6.7. Electronic data capture

Observations/measurements in the study were recorded electronically using the following program s:

REES Centron Environmental Monitoring system version SQL 2.0 (REES scientific, Trenton, NJ,

USA) ; TOXDATA version 8.0 (WIL Research Europe B.V., ‘s

- Hertogenbosch, The Netherlands): Clinical

signs, Body weights.

6.8. Interpretation

The LD50 w as estimated as exceeding 2000 mg/kg b.w.

The results were evaluated according to the Globally Harmonized System of Classification and

Labelling of Chemicals (GHS) of the United Nations and the EC criteria for classification and labelling

of dangerous substances and preparations (Council Directive 67/548/EEC and all adaptations to

technical progress and amendments of this Directive published in the Official Journal of the European

Union ).

6.9. List of deviations

6.9.1. List of protocol deviations

1. The animals were identified with an ear and tail mark.

Evaluation: The identification method was suitable for this study

.

The study integrity was not adversely affected by the deviations.

6.9.2. List of standard operating procedures deviations

Any deviations from standard operating procedures were evaluated and filed in the study file. There

were no deviations from standard operating procedures that affected the integrity of the study.1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 11

- 7. RESULTS

7.1. Mortality ( Table 1 )

No mortality occurred after dosing 5 females at 2000 mg/kg

.

7.2. Clinical s igns ( Table 2 )

Hunched posture and/or piloerection were seen in two females

7.3. Body w eights ( Table 3 )

The body weight gain shown by the animals over the study period was considered to be similar to that

expected of normal untreated animals of the same age and strain

.

7.4. Macroscopic f indings ( Table 4 )

No abnormalities were found at macroscopic post mortem examination of the animals.

8. CONCLUSION

The oral LD50 value of 1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) in

Wistar rats was established to exceed 2000 mg/kg body weight.

Based on these results , 1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1)

does not have to be classified and has no obligatory labelling requirement for acute oral toxicity

according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) of

the United Nations ( 2011 ) and Regulation (EC) No 1272/2008 on classification, labelling and

packaging of substances and mixtures

.1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 12

- TABLE 1 MORTALITY DATA

TEST DAY

HOURS AFTER TREATMENT 1

0 1

2 1

4 2 3 4 5 6 7 8 9 10 11 12 13 14 15

FEMALES 2000 MG/KG

- - - - - - - - - - - - - - - - - FEMALES 2000 MG/KG

- - - - - - - - - - - - - - - - -

TABLE 2 CLINICAL SIGNS

TEST DAY

HOURS AFTER TREATMENT MAX

GRADE 1

0 1

2 1

4 2 3 4 5 6 7 8 9 10 11 12 13 14 15

FEMALES 2000 MG/KG

ANIMAL 1

Posture

Hunched posture (1)

- 1 1

- - - - - - - - - - - - - - Skin / fur

Piloerection (1)

- 1 1

- - - - - - - - - - - - - -

FEMALES 2000 MG/KG

ANIMAL 2

Posture

Hunched posture (1) 1

- - - - - - - - - - - - - - - -

ANIMAL 3

No clinical signs noted

- - - - - - - - - - - - - - - - -

ANIMAL 4

No clinical signs noted

- - - - - - - - - - - - - - - - - ANIMAL 5

No clinical signs noted

- - - - - - - - - - - - - - - - -

- = Sign not observed

TABLE 3 BODY WEIGHTS (GRAM)

SEX/DOSE LEVEL ANIMAL DAY 1 DAY 8 DAY 15

FEMALES 2000 MG/KG

1 153 184 198

MEAN 153 184 198

ST.DEV.

--- --- ---

N 1 1 1

FEMALES 2000 MG/KG

2 167 195 201

3 184 219 229

4 186 212 218

5 190 222 246

MEAN 182 212 224

ST.DEV. 10 12 19

N 4 4 41- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 13

- TABLE 4 MACROSCOPIC FINDINGS

ANIMAL ORGAN FINDING DAY OF DEATH

FEMALES 2000 MG/KG

1 No findings noted Scheduled necropsy

Day 15 after treatment

FEMALES 2000 MG/KG

2 No findings n oted Scheduled necropsy

Day 15 after treatment

3 No findings noted Scheduled necropsy

Day 15 after treatment

4 No findings noted Scheduled necropsy

Day 15 after treatment

5 No findings noted Scheduled necrops y

Day 15 after treatment1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 14

- APPENDIX 1 : TEST SUBSTANCE CERTIFICATE OF ANALY SIS1- Propanesulfonic acid, 2

- hydroxy -3- (2

- propen -1- yloxy)

- , sodium salt (1:1) Project 500679

- Page 15

-