FINAL REPORT

Study Title

EVALUATION OF THE MUTAGENIC ACTIVITY OF 1-

PROPANESULFONIC ACID, 2-HYDROXY-3-(2-PROPEN-1-

YLOXY)-, SODIUM SALT (1:1) IN THE SALMONELLA 

TYPHIMURIUM REVERSE MUTATION ASSAY AND THE 

ESCHERICHIA COLI REVERSE MUTATION ASSAY

(WITH INDEPENDENT REPEAT)

Author

C.M. Verspeek-Rip

Test Facility

WIL Research Europe B.V.

Hambakenwetering 7

5231 DD ‘s-Hertogenbosch

The Netherlands

 

Laboratory Project Identification

Project 500680

Substance 204240/A

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1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

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1. CONTENTS

1. CONTENTS.................................................................................................................................. 2

2. STATEMENT OF GLP COMPLIANCE.......................................................................................... 3

3. QUALITY ASSURANCE STATEMENT......................................................................................... 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 METHODS ...................................................................................................... 7

6.1. Test substance...................................................................................................................... 7

6.1.1. Test substance information ......................................................................................... 7

6.1.2. Study specific test substance information.................................................................... 7

6.1.3. Test substance preparation......................................................................................... 8

6.2. Reference substances .......................................................................................................... 8

6.2.1. Negative control .......................................................................................................... 8

6.2.2. Positive controls .......................................................................................................... 8

6.3. Test system........................................................................................................................... 8

6.4. Cell culture............................................................................................................................ 9

6.5. Metabolic activation system .................................................................................................. 9

6.5.1. Preparation of S9-fraction ........................................................................................... 9

6.5.2. Preparation of S9-mix ................................................................................................10

6.6. Study design ........................................................................................................................10

6.6.1. Dose range finding test ..............................................................................................10

6.6.2. Mutation assay...........................................................................................................10

6.6.3. Colony counting .........................................................................................................11

6.7. Electronic data capture.........................................................................................................11

6.8. Interpretation........................................................................................................................11

6.8.1. Acceptability of the assay...........................................................................................11

6.8.2. Data evaluation and statistical procedures .................................................................11

6.9. List of deviations ..................................................................................................................12

6.9.1. List of protocol deviations...........................................................................................12

6.9.2. List of standard operating procedures deviations .......................................................12

7. RESULTS....................................................................................................................................12

7.1. Dose range finding test ........................................................................................................12

7.2. Mutation assay.....................................................................................................................13

8. DISCUSSION AND CONCLUSION .............................................................................................13

9. REFERENCES............................................................................................................................13

TABLES

Table 1 Experiment 1: Mutagenic response of 1-Propanesulfonic acid, 2-hydroxy-3-

(2-propen-1-yloxy)-, sodium salt (1:1) in the Salmonella typhimurium reverse 

mutation assay and in the Escherichia coli reverse mutation assay ...........................14

Table 2 Experiment 2: Mutagenic response of 1-Propanesulfonic acid, 2-hydroxy-3-

(2-propen-1-yloxy)-, sodium salt (1:1) in the Salmonella typhimurium reverse 

mutation assay and in the Escherichia coli reverse mutation assay ...........................15

APPENDICES 

APPENDIX 1 SUMMARY TABLES..................................................................................................14

APPENDIX 2 SUPPORTING MATERIALS AND METHOD..............................................................16

APPENDIX 3 DETAILED TABLES...................................................................................................17

APPENDIX 4 HISTORICAL CONTROL DATA OF THE SOLVENT CONTROL ...............................27

APPENDIX 5 HISTORICAL CONTROL DATA OF THE POSITIVE CONTROL SUBSTANCES.......28

APPENDIX 6 CERTIFICATE OF ANALYSIS ...................................................................................29

 

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4. SUMMARY

Evaluation of the mutagenic activity of 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, 

sodium salt (1:1) in the Salmonella typhimurium reverse mutation assay and the Escherichia coli 

reverse mutation assay (with independent repeat).

1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) was tested in the 

Salmonella typhimurium reverse mutation assay with four histidine-requiring strains of Salmonella 

typhimurium (TA1535, TA1537, TA98 and TA100) and in the Escherichia coli reverse mutation assay 

with a tryptophan-requiring strain of Escherichia coli (WP2uvrA). The test was performed in two 

independent experiments in the presence and absence of S9-mix (rat liver S9-mix induced by a 

combination of phenobarbital and ß-naphthoflavone).

The study procedures described in this report were based on the most recent OECD and EC 

guidelines.

Batch 12E1101 of 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) was a 

clear colourless liquid. 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) was 

dissolved in Milli-Q water. 

In the dose range finding test, 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt 

(1:1) was tested up to concentrations of 5000 μg/plate in the absence and presence of S9-mix in the 

strains TA100 and WP2uvrA. 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt 

(1:1) did not precipitate on the plates at this dose level. The bacterial background lawn was not 

reduced at any of the concentrations tested and no biologically relevant decrease in the number of 

revertants was observed. Results of this dose range finding test were reported as part of the first 

experiment of the mutation assay.

Based on the results of the dose range finding test, 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-

yloxy)-, sodium salt (1:1) was tested in the first mutation assay at a concentration range of 100 to 

5000 μg/plate in the absence and presence of 5% (v/v) S9-mix in tester strains TA1535, TA1537 and 

TA98. In an independent repeat of the assay with additional parameters, 1-Propanesulfonic acid, 2-

hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) was tested at the same concentration range as the 

first assay in the absence and presence of 10% (v/v) S9-mix in tester strains TA1535, TA1537, TA98, 

TA100 and WP2uvrA. The bacterial background lawn was not reduced at any of the concentrations 

tested and no biologically relevant decrease in the number of revertants was observed. 

1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) did not induce a significant 

dose-related increase in the number of revertant (His+

) colonies in each of the four tester strains 

(TA1535, TA1537, TA98 and TA100) and in the number of revertant (Trp+

) colonies in tester strain 

WP2uvrA both in the absence and presence of S9-metabolic activation. These results were confirmed 

in an independently repeated experiment.

In this study, the negative and strain-specific positive control values were within the laboratory 

historical control data ranges indicating that the test conditions were adequate and that the metabolic 

activation system functioned properly.

Based on the results of this study it is concluded that 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen1-yloxy)-, sodium salt (1:1) is not mutagenic in the Salmonella typhimurium reverse mutation assay 

and in the Escherichia coli reverse mutation assay. 

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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

5231 DD ‘s-Hertogenbosch

The Netherlands

Study Director C.M. Verspeek-Rip

Study Plan Start : 04 February, 2013

Completion : 14 February, 2013

5.2. Aims of the study

The objective of this study was to evaluate the test substance for its ability to induce reverse mutations 

in a gene of histidine-requiring Salmonella typhimurium bacterial strains resulting in histidineindependent strains, and in a gene of tryptophan-requiring Escherichia coli bacterial strain resulting in 

a tryptophan-independent strain.

Background of the test system

The Salmonella typhimurium reverse mutation assay and the Escherichia coli reverse mutation assay 

have been shown to be rapid and adequate indicators for the mutagenic activity of a wide range of 

chemical compounds.

The assay was conducted in the absence and presence of a metabolizing system (S9-mix). 

The Salmonella typhimurium strains used in this study were TA1535, TA1537, TA98 and TA100. The 

Escherichia coli strain used was WP2uvrA. The strains TA1537 and TA98 are capable of detecting 

frameshift mutagens, strains TA1535, TA100 and WP2uvrA are capable of detecting base-pair 

substitution mutagens (1-5).

5.3. Guidelines

The study procedures described in this report were based on the following guidelines:

- Organisation for Economic Co-operation and Development (OECD), OECD Guidelines for Testing 

of Chemicals; Guideline no. 471: "Genetic Toxicology: Bacterial Reverse Mutation Test" (Adopted 

July 21, 1997).

- European Community (EC). Commission regulation (EC) No. 440/2008, Part B: Methods for the 

Determination of Toxicity and other health effects, Guideline B.13/14: "Mutagenicity: Reverse 

Mutation Test using Bacteria”. Official Journal of the European Union No. L142, 31 May 2008.

- Code of Federal Regulations, Title 40 – Protection of Environment. Section 799.9510 Toxic 

Substances Control Act (TSCA) bacterial reverse mutation test. 

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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. 

6. MATERIALS AND METHODS 

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

Structure

O SO3

-

OH

Na+

Molecular formula C6H11NaO5S

Molecular weight 218.22

CAS Number 52556-42-0

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 6; Certificate of analysis1

. 

6.1.2. Study specific test substance information

Purity/composition correction factor 

required

No 

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

maximum duration: 1 hour

Stability in vehicle Water: 1 year

Solubility in vehicle Water: 1-45%

1

 The C of A provided for Lot No. 12E1101 is correct as originally provided. The value reported as Dry Residue is made up of 

two components which together add up to the 40.9 % reported as Dry Residue on the C of A. When doing the Dry Residue test 

only the water (59.1%) volatilizes, the sodium-1-alloxy-2-hydroxypropane sulphonate and the 3-allyl-1,2-propanediol together 

comprise the Dry Residue, respectively 94.5% and 5.5%. On the basis of the entire sample (water present) this pecentage will 

be 38.64 and 2.26 as reported previously.

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6.1.3. Test substance preparation

No correction was made for the purity/composition of the test compound. 

The test substance was dissolved in Milli-Q water (Millipore Corp., Bedford, MA., USA). The stock 

solution was filter (0.22 μm)-sterilized. Test substance concentrations were used within 2.5 hours after 

preparation.

6.2. Reference substances

6.2.1. Negative control

The vehicle of the test substance, which was Milli-Q water. 

6.2.2. Positive controls

Without metabolic activation (-S9-mix):

Strain Chemical Concentration/plate Solvent

TA1535 sodium azide (SA) 5 μg Saline

(Sigma Aldrich Chemie, Steinheim, Germany) 

TA1537 ICR-191 (Acros Organics, Geel, Belgium) 2.5 μg DMSO

TA98 2-nitrofluorene (NF) (Merck) 10 μg DMSO

TA100 methylmethanesulfonate (MMS) (Sigma) 650 μg DMSO

WP2uvrA 4-nitroquinoline N-oxide (4-NQO) (Sigma) 10 μg DMSO

With metabolic activation (+S9-mix):

The positive control substance used for all tester strains was 2-aminoanthracene (2AA) (Sigma). The 

following concentrations were used:

Strain Concentration/plate Amount of S9-mix Solvent

TA1535 2.5 μg 5 and 10% DMSO

TA1537 2.5 μg 5% DMSO 

TA1537 5 μg 10% DMSO 

TA98 1 μg 5 and 10% DMSO

TA100 1 μg 5% DMSO

TA100 2.5 μg 10% DMSO

WP2uvrA 10 μg 5 and 10% DMSO

Solvents for reference substances

Saline = physiological saline (B. Braun, Melsungen AG, Germany)

DMSO = dimethyl sulfoxide (Merck) 

6.3. Test system

Test system Salmonella typhimurium bacteria and Escherichia coli bacteria

Rationale Recommended test system in international guidelines (e.g. OECD, EC).

Source Trinova Biochem GmbH, Germany (Master culture from Dr. Bruce N. Ames) 

(TA1535: 2006, TA1537: 2009, TA98: 2006, TA100: 2006) and (Master culture 

from The National Collections of Industrial and Marine Bacteria, Aberdeen, UK) 

(WP2uvrA, 2008)

The characteristics of the different Salmonella typhimurium strains were as follows:

Strain Histidine mutation Mutation type

TA1537 hisC3076 Frameshift

TA98 hisD3052/R-factor* Frameshift

TA1535 hisG46 Base-pair substitutions

TA100 hisG46/R-factor* Base-pair substitutions

*: R-factor = plasmid pKM101 (increases error-prone DNA repair)

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Each tester strain contained the following additional mutations:

rfa : deep rough (defective lipopolysaccharide cellcoat)

gal : mutation in the galactose metabolism

chl : mutation in nitrate reductase

bio : defective biotin synthesis

uvrB : loss of the excision repair system (deletion of the ultraviolet-repair B gene)

The Salmonella typhimurium strains are regularly checked to confirm their histidine-requirement, 

crystal violet sensitivity, ampicillin resistance (TA98 and TA100), UV-sensitivity and the number of 

spontaneous revertants.

The Escherichia coli WP2uvrA strain detects base-pair substitutions. The strain lacks an excision repair 

system and is sensitive to agents such as UV. The sensitivity of the strain to a wide variety of 

mutagens has been enhanced by permeabilization of the strain using Tris-EDTA treatment (Ref.1). 

The strain is regularly checked to confirm the tryptophan-requirement, UV-sensitivity and the number 

of spontaneous revertants.

Stock cultures of the five strains were stored in liquid nitrogen (-196°C).

6.4. Cell culture

Preparation of bacterial cultures

Samples of frozen stock cultures of bacteria were transferred into enriched nutrient broth (Oxoid LTD, 

Hampshire, England) and incubated in a shaking incubator (37°C, 150 spm), until the cultures reached 

an optical density of 1.0 ± 0.1 at 700 nm (109 cells/ml). Freshly grown cultures of each strain were 

used for a test.

Agar plates

Agar plates (ø 9 cm) contained 25 ml glucose agar medium. Glucose agar medium contained per liter: 

18 g purified agar (Oxoid LTD) in Vogel-Bonner Medium E, 20 g glucose (Fresenius Kabi, Bad 

Homburg, Germany). The agar plates for the test with the Salmonella typhimurium strains also 

contained 12.5 μg/plate biotin (Merck) and 15 μg/plate histidine (Merck) and the agar plates for the 

test with the Escherichia coli strain contained 15 μg/plate tryptophan (Acros Organics).

Top agar

Milli-Q water containing 0.6% (w/v) bacteriological agar (Oxoid LTD) and 0.5% (w/v) sodium chloride 

(Merck) was heated to dissolve the agar. Samples of 3 ml top agar were transferred into 10 ml glass 

tubes with metal caps. Top agar tubes were autoclaved for 20 min at 121 ± 3°C.

 

Environmental conditions

All incubations were carried out in a controlled environment at a temperature of 37.0 ± 1.0°C (actual 

range 36.4 – 39.8°C) in the dark. Temporary deviations of maximally 2 hours (in the range of 

38.0 – 39.0°C) occurred due to addition of plates (which were at room temperature) to the incubator or 

due to opening and closing the incubator door. Based on laboratory historical data these deviations 

are considered not to affect the study integrity. The temporary deviation from the temperature (with a 

maximum of 39.8°C) is explained in protocol deviation 1. 

6.5. Metabolic activation system

Rat liver microsomal enzymes were routinely prepared from adult male Wistar rats, which were 

obtained from Charles River, Sulzfeld, Germany.

6.5.1. Preparation of S9-fraction

The animals were housed at WIL Research Europe in a special room under standard laboratory 

conditions, as described in the Standard Operating Procedures. The rats were orally dosed for three 

consecutive days with a suspension of phenobarbital (Bufa B.V., IJsselstein, The Netherlands) 

(80 mg/kg body weight) and ß-naphthoflavone (Sigma Aldrich Chemie) (100 mg/kg body weight) in 

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corn oil (Roth, Karlsruhe, Germany) (they were denied access to food for 3 to 4 hours preceding each 

dosing). One day after the final exposure (24 h), the rats were sedated using oxygen/carbon dioxide 

and then killed by decapitation. The rats received a limited quantity of food during the night before 

sacrifice. The livers of the rats were removed aseptically, and washed in cold (0°C) sterile 0.1 M 

sodium phosphate buffer (pH 7.4) containing 0.1 mM Na2-EDTA (Merck). Subsequently the livers were 

minced in a blender and homogenized in 3 volumes of phosphate buffer with a Potter homogenizer. 

The homogenate was centrifuged for 15 min at 9000 g. The supernatant (S9) was transferred into 

sterile ampoules, which were stored in liquid nitrogen (-196°C) for a maximum of 1 year.

Before use, all S9 batches were characterised with the mutagens Benzo-(a)-pyrene (Sigma) and 

2-aminoanthracene (Sigma), which require metabolic activation, in tester strain TA98 at concentrations 

of 5 μg/plate and 1 μg/plate, respectively.

6.5.2. Preparation of S9-mix

S9-mix was prepared immediately before use and kept on ice. S9-mix contained per 10 ml: 30 mg 

NADP (Randox Laboratories Ltd., Crumlin, United Kingdom) and 15.2 mg glucose-6-phosphate 

(Roche Diagnostics, Mannheim, Germany) in 5.5 ml or 5.0 ml Milli-Q water (first or second experiment 

respectively) (Millipore Corp., Bedford, MA., USA); 2 ml 0.5 M sodium phosphate buffer pH 7.4; 1 ml 

0.08 M MgCl2 solution (Merck); 1 ml 0.33 M KCl solution (Merck). The above solution was filter 

(0.22 μm)-sterilized. To 9.5 ml of S9-mix components 0.5 ml S9-fraction was added (5% (v/v) 

S9-fraction) to complete the S9-mix in the first experiment and to 9.0 ml of S9-mix components 1.0 ml 

S9-fraction was added (10% (v/v) S9-fraction) to complete the S9-mix in the second experiment. 

6.6. Study design

6.6.1. Dose range finding test

Selection of an adequate range of doses was based on a dose range finding test with the strains 

TA100 and WP2uvrA, both with and without 5% (v/v) S9-mix. Eight concentrations, 3, 10, 33, 100, 333, 

1000, 3330 and 5000 μg/plate were tested in triplicate. This dose range finding test was reported as a 

part of the first experiment of the mutation assay. The highest concentration of 1-Propanesulfonic acid, 

2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) used in the subsequent mutation assay was 

5000 μg/plate. 

6.6.2. Mutation assay

At least four different doses (increasing with approximately half-log steps, see protocol deviation 2) of 

the test substance were tested in triplicate in each strain. In the first experiment 1-Propanesulfonic 

acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) was tested both in the absence and presence 

of 5% (v/v) S9-mix in tester strains TA1535, TA1537 and TA98. In an independent repeat of the assay 

with additional parameters, the test substance was tested both in the absence and presence of 

10% (v/v) S9-mix in all tester strains.

The negative control (vehicle) and relevant positive controls were concurrently tested in each strain in 

the presence and absence of S9-mix.

Top agar in top agar tubes was melted by heating to 45°C. The following solutions were successively 

added to 3 ml molten top agar: 0.1 ml of a fresh bacterial culture (109

cells/ml) of one of the tester 

strains, 0.1 ml of a dilution of the test substance in Milli-Q water and either 0.5 ml S9-mix (in case of 

activation assays) or 0.5 ml 0.1 M phosphate buffer (in case of non-activation assays). The ingredients 

were mixed on a Vortex and the content of the top agar tube was poured onto a selective agar plate. 

After solidification of the top agar, the plates were inverted and incubated in the dark at 37.0 ± 1.0 °C 

for 48 ± 4 h. After this period revertant colonies (histidine independent (His+

) for Salmonella 

typhimurium bacteria and tryptophan independent (Trp+

) for Escherichia coli were counted.

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6.6.3. Colony counting

The revertant colonies (histidine independent or tryptophan independent) were counted manually if 

less than 40 colonies per plate were present. If more than 40 colonies were present, these were

counted automatically with a Biocount 4000 Pro-S-colony counter. Plates with abundant test 

substance precipitate which interfered with automated colony counting were counted manually and the 

evidence of test substance precipitate on the plates was recorded. The condition of the bacterial 

background lawn was evaluated, both macroscopically and microscopically by using a dissecting 

microscope.

6.7. Electronic data capture

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

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

USA): Temperature. 

6.8. Interpretation

6.8.1. Acceptability of the assay

A Salmonella typhimurium reverse mutation assay and/or Escherichia coli reverse mutation assay is 

considered acceptable if it meets the following criteria:

a) The negative control data (number of spontaneous revertants per plate) should be within the 

laboratory historical range for each tester strain (see APPENDIX 4). 

b) The positive control chemicals should produce responses in all tester strains, which are within the 

laboratory historical range documented for each positive control substance. Furthermore, the mean 

plate count should be at least three times the concurrent vehicle control group mean (see 

APPENDIX 5). 

c) The selected dose range should include a clearly toxic concentration or should exhibit limited 

solubility as demonstrated by the preliminary toxicity range-finding test or should extend to 

5 mg/plate.

6.8.2. Data evaluation and statistical procedures

No formal hypothesis testing was done.

A test substance is considered negative (not mutagenic) in the test if:

a) The total number of revertants in tester strain TA100 is not greater than two (2) times the 

concurrent control, and the total number of revertants in tester strains TA1535, TA1537, TA98 or 

WP2uvrA is not greater than three (3) times the concurrent vehicle control.

b) The negative response should be reproducible in at least one independently repeated experiment.

A test substance is considered positive (mutagenic) in the test if:

a) The total number of revertants in tester strain TA100 is greater than two (2) times the concurrent 

control, or the total number of revertants in tester strains TA1535, TA1537, TA98 or WP2uvrA is 

greater than three (3) times the concurrent vehicle control. 

b) In case a repeat experiment is performed when a positive response is observed in one of the tester 

strains, the positive response should be reproducible in at least one independently repeated 

experiment.

The preceding criteria were not absolute and other modifying factors might enter into the final 

evaluation decision.

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6.9. List of deviations

6.9.1. List of protocol deviations

1. The temperature was recorded to be outside the range of 37.0 ± 1.0°C as specified in the 

protocol for approximately 2 hours in the first mutation assay (with a maximum of 39.8°C).

Evaluation: This short term deviation was observed within three hours after initiation of the test 

and was caused by adjustment of the temperature in the incubator after placing an amount of 

selective agar plates in the incubator. The negative control data (number of spontaneous 

revertants per plate) were within the laboratory historical range for each tester strain, therefore 

this short deviation of the temperature had no effect on the results of the study.

2. In the second mutation assay in tester strain TA100 in the absence of S9-mix, four dose levels

were tested instead of five dose levels as specified in the protocol. 

Evaluation: No bacteria were added to the lowest dose level of 100 μg/plate. Since the four 

highest dose levels were tested, and no toxicity and/or precipitate was observed, testing of a 

lower dose level would have given no additional information. Therefore this deviation had no 

influence on the study result.

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. 

7. RESULTS

7.1. Dose range finding test 

1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) was tested in the tester 

strains TA100 and WP2uvrA with concentrations of 3, 10, 33, 100, 333, 1000, 3330 and 5000 μg/plate 

in the absence and presence of S9-mix.

This dose range finding test is reported as a part of the first experiment of the mutation test (Table 1). 

The individual data are presented in APPENDIX 3.

Precipitate

Precipitation of 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) on the 

plates was not observed at the start or at the end of the incubation period in both tester strains. 

Toxicity

To determine the toxicity of 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1), 

the reduction of the bacterial background lawn, the increase in the size of the microcolonies and the 

reduction of the revertant colonies were examined. The definitions are stated in APPENDIX 2.

No reduction of the bacterial background lawn and no biologically relevant decrease in the number of 

revertants were observed.

Mutagenicity 

In the dose range finding test, no increase in the number of revertants was observed upon treatment 

with 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) under all conditions 

tested.

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7.2. Mutation assay 

Based on the results of the dose range finding test, 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-

yloxy)-, sodium salt (1:1) was tested up to concentrations of 5000 μg/plate in the absence and 

presence of S9-mix in two mutation assays. The first mutation experiment was performed with the 

strains TA1535, TA1537 and TA98 and the second mutation experiment was performed with the 

strains TA1535, TA1537, TA98, TA100 and WP2uvrA. The results are shown in Table 1 and Table 2,

the individual data are presented in APPENDIX 3.

Precipitate

Precipitation of 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) on the 

plates was not observed at the start or at the end of the incubation period. 

Toxicity 

In both mutation assays, there was no reduction of the bacterial background lawn and no biologically 

relevant decrease in the number of revertants at any of the concentrations tested in all tester strains in 

the absence and presence of S9-mix.

Mutagenicity 

In both mutation assays, no increase in the number of revertants was observed upon treatment with 

1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) under all conditions tested.

8. DISCUSSION AND CONCLUSION

All bacterial strains showed negative responses over the entire dose range, i.e. no significant doserelated increase in the number of revertants in two independently repeated experiments. 

The negative and strain-specific positive control values were within the laboratory historical control 

data ranges indicating that the test conditions were adequate and that the metabolic activation system 

functioned properly.

Based on the results of this study it is concluded that 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen1-yloxy)-, sodium salt (1:1) is not mutagenic in the Salmonella typhimurium reverse mutation assay 

and in the Escherichia coli reverse mutation assay.

9. REFERENCES

1 Leonardo, J.M., Dornfeld, S.S. and Peak, M.J., 1984, Evaluation of E. coli K12 343 \ 113 and 

derived strains for microbial mutagenicity assays. Mutation Res., 130, 87-95.

2 Ames, B.N., McCann, J. and Yamasaki, E., 1975, Methods for detecting carcinogens and 

mutagens with the Salmonella/mammalian microsome mutagenicity test, Mutation Res., 31, 347-

364.

3 Maron, D.M. and Ames, B.N., 1983, Revised methods for the Salmonella mutagenicity test, 

Mutation Res., 113, 173-215. Erratum, 1983, Mutation Res., 113, 533.

4 Green, M.H.L. and Muriel, W.J., 1976, Mutagen testing using Trp+ reversion in Escherichia coli, 

Mutation Res., 38, 3-32.

5 Vogel, H.J. and Bonner, D.M., 1956, Acetylornithinase of Escherichia coli: partial purification and 

some properties. J. Biol. Chem., 218, 97-106.

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 14 - 

APPENDIX 1 SUMMARY TABLES

Table 1 Experiment 1: Mutagenic response of 1-Propanesulfonic acid, 2-hydroxy-3-(2-

propen-1-yloxy)-, sodium salt (1:1) in the Salmonella typhimurium reverse 

mutation assay and in the Escherichia coli reverse mutation assay

Dose Mean number of revertant colonies/3 replicate plates (± S.D.) with

(μg/plate) different strains of Salmonella typhimurium and one Escherichia coli strain

TA1535 TA1537 TA98 TA100 WP2uvrA

Without S9-mix

 positive control 949 ± 46 538 ± 16 934 ± 49 702 ± 26 1185 ± 82

 solvent control 13 ± 5 6 ± 1 17 ± 2 104 ± 6 28 ± 2 

 3 104 ± 14 36 ± 4 

 10 97 ± 12 33 ± 2 

 33 94 ± 17 32 ± 4 

 100 13 ± 3 6 ± 1 16 ± 3 99 ± 13 34 ± 7 

 333 10 ± 6 6 ± 2 17 ± 4 105 ± 6 28 ± 3 

 1000 11 ± 1 5 ± 1 17 ± 2 104 ± 14 31 ± 4 

 3330 10 ± 3 8 ± 1 18 ± 2 115 ± 2 39 ± 6 

 5000 13 ± 3 7 ± 2 19 ± 5 91 ± 6 30 ± 4 

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

With S9-mix1

 positive control 410 ± 6 478 ± 20 785 ± 93 1048 ± 52 458 ± 25

 solvent control 13 ± 3 8 ± 1 24 ± 2 116 ± 8 35 ± 8 

 3 97 ± 21 41 ± 1 

 10 105 ± 16 39 ± 1 

 33 106 ± 5 31 ± 1 

 100 13 ± 1 7 ± 2 23 ± 5 94 ± 11 29 ± 4 

 333 8 ± 4 7 ± 1 25 ± 5 109 ± 14 30 ± 1 

 1000 9 ± 3 7 ± 3 27 ± 5 98 ± 13 30 ± 2 

 3330 11 ± 4 5 ± 4 27 ± 4 100 ± 10 40 ± 5 

 5000 12 ± 5 6 ± 3 25 ± 2 106 ± 6 31 ± 3 

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

Solvent control: 0.1 ml Milli-Q water

1 The S9-mix contained 5% (v/v) S9 fraction

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 15 - 

APPENDIX 1 – continued –

Table 2 Experiment 2: Mutagenic response of 1-Propanesulfonic acid, 2-hydroxy-3-(2-

propen-1-yloxy)-, sodium salt (1:1) in the Salmonella typhimurium reverse 

mutation assay and in the Escherichia coli reverse mutation assay

Dose Mean number of revertant colonies/3 replicate plates (± S.D.) with

(μg/plate) different strains of Salmonella typhimurium and one Escherichia coli strain

TA1535 TA1537 TA98 TA100 WP2uvrA

Without S9-mix

 positive control 822 ± 22 598 ± 4 1056 ± 105 743 ± 49 1024 ± 64

 solvent control 11 ± 2 6 ± 2 15 ± 3 122 ± 18 26 ± 7 

 100 7 ± 4 5 ± 2 19 ± 4 2) 19 ± 3 

 333 10 ± 2 5 ± 3 19 ± 2 125 ± 6 18 ± 3 

 1000 10 ± 2 4 ± 1 21 ± 2 137 ± 33 17 ± 4 

 3330 10 ± 3 4 ± 1 21 ± 1 130 ± 11 23 ± 2 

 5000 10 ± 4 3 ± 2 19 ± 4 119 ± 11 15 ± 3 

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

With S9-mix1

 positive control 183 ± 21 474 ± 5 373 ± 39 339 ± 6 316 ± 33

 solvent control 9 ± 1 4 ± 1 27 ± 7 109 ± 24 26 ± 7 

 100 7 ± 3 5 ± 0 28 ± 2 128 ± 3 27 ± 4 

 333 7 ± 3 8 ± 3 21 ± 2 152 ± 19 27 ± 4 

 1000 7 ± 3 6 ± 3 28 ± 4 132 ± 15 33 ± 5 

 3330 6 ± 1 6 ± 3 27 ± 5 116 ± 13 32 ± 2 

 5000 10 ± 1 4 ± 2 28 ± 3 124 ± 11 17 ± 3 

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

Solvent control: 0.1 ml Milli-Q water

1 The S9-mix contained 10% (v/v) S9 fraction

2 Not determined

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 16 - 

APPENDIX 2 SUPPORTING MATERIALS AND METHOD

Bacterial background lawn evaluation

The condition of the bacterial background lawn is evaluated (if indicated), both macroscopically and 

microscopically by using a dissecting microscope (results are normal unless indicated in tables).

Definition Characteristics

Normal Distinguished by a healthy microcolony lawn.

Slightly reduced Distinguished by a slight thinning of the microcolony lawn.

Moderately reduced Distinguished by a moderate thinning of the microcolony lawn.

Extremely reduced Distinguished by an extreme thinning of the microcolony lawn and an 

increase in the size of the microcolonies compared to the solvent control 

plate. 

Absent Distinguished by a complete lack of any microcolony background lawn.

Precipitation evaluation

Evidence of test article precipitate on the plates is recorded by addition of the following precipitation 

definition.

Definition Characteristics

Slight Precipitate Distinguished by noticeable precipitate on the plate.

However, the precipitate does not influence automated counting of the plate.

Moderate Precipitate Distinguished by a marked amount of precipitate on the plate, requiring the 

plate to be hand counted.

Heavy Precipitate Distinguished by a large amount of precipitate on the plate, making the 

required hand count difficult.

Evaluation of the reduction in the number of revertants

The reduction in the number of revertant colonies compared to number of revertants in the solvent 

control is evaluated as follows:

A reduction of 21-40%: slight reduction.

A reduction of 41-60%: moderate reduction.

A reduction of 61-99%: extreme reduction.

If the size of the microcolonies was increased to small colonies due to an extremely reduced 

background lawn the reduction is evaluated as microcolonies. If no revertant colonies are observed on 

the plates the reduction is evaluated as a complete lack of revertants.

However, any mean plate count equal to the minimal value of the historical control data range should 

be considered not toxic.

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 17 - 

APPENDIX 3 DETAILED TABLES

Individual plate counts; (following pages)

Experiment 1

Strain TA1535

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

 WITHOUT S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 934 912 1001 949 ± 46

 solvent control 19 12 9 13 ± 5 

 100 10 14 16 13 ± 3 

 333 5 17 7 10 ± 6 

 1000 12 10 12 11 ± 1 

 3330 7 10 13 10 ± 3 

 5000 15 14 9 13 ± 3 

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

 WITH S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 406 417 407 410 ± 6 

 solvent control 16 10 12 13 ± 3 

 100 14 13 13 13 ± 1 

 333 7 12 4 8 ± 4 

 1000 7 12 7 9 ± 3 

 3330 12 14 6 11 ± 4 

 5000 10 8 17 12 ± 5 

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

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APPENDIX 3 – continued –

Experiment 1

Strain TA1537

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

 WITHOUT S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 527 530 556 538 ± 16

 solvent control 7 7 5 6 ± 1 

 100 6 5 7 6 ± 1 

 333 8 4 5 6 ± 2 

 1000 5 5 4 5 ± 1 

 3330 9 9 7 8 ± 1 

 5000 7 8 5 7 ± 2 

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

 WITH S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 496 456 483 478 ± 20

 solvent control 8 7 9 8 ± 1 

 100 6 7 9 7 ± 2 

 333 6 8 7 7 ± 1 

 1000 4 7 9 7 ± 3 

 3330 5 9 2 5 ± 4 

 5000 3 9 6 6 ± 3 

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

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1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

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APPENDIX 3 – continued –

Experiment 1

Strain TA98

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

 WITHOUT S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 912 990 901 934 ± 49

 solvent control 19 16 17 17 ± 2 

 100 13 17 18 16 ± 3 

 333 14 15 22 17 ± 4 

 1000 17 15 18 17 ± 2 

 3330 19 19 16 18 ± 2 

 5000 20 24 14 19 ± 5 

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

 WITH S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 823 679 854 785 ± 93

 solvent control 25 21 25 24 ± 2 

 100 28 22 19 23 ± 5 

 333 19 27 28 25 ± 5 

 1000 33 23 25 27 ± 5 

 3330 27 24 31 27 ± 4 

 5000 24 27 25 25 ± 2 

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

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1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

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APPENDIX 3 – continued –

Experiment 1

Strain TA100

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

 WITHOUT S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 708 724 673 702 ± 26

 solvent control 109 98 106 104 ± 6 

 3 96 120 95 104 ± 14

 10 92 111 88 97 ± 12

 33 110 95 76 94 ± 17

 100 107 105 84 99 ± 13

 333 109 98 109 105 ± 6 

 1000 95 120 98 104 ± 14

 3330 115 113 117 115 ± 2 

 5000 86 97 90 91 ± 6 

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

 WITH S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 996 1099 1050 1048 ± 52

 solvent control 108 123 116 116 ± 8 

 3 76 118 96 97 ± 21

 10 124 94 98 105 ± 16

 33 101 110 106 106 ± 5 

 100 107 85 91 94 ± 11

 333 118 116 93 109 ± 14

 1000 103 107 83 98 ± 13

 3330 109 90 101 100 ± 10

 5000 99 111 107 106 ± 6 

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

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1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

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APPENDIX 3 – continued –

Experiment 1

Strain WP2uvrA

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

 WITHOUT S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 1098 1197 1261 1185 ± 82

 solvent control 26 28 30 28 ± 2 

 3 32 37 39 36 ± 4 

 10 35 33 32 33 ± 2 

 33 35 28 32 32 ± 4 

 100 40 27 35 34 ± 7 

 333 28 25 31 28 ± 3 

 1000 34 33 27 31 ± 4 

 3330 34 39 45 39 ± 6 

 5000 26 29 34 30 ± 4 

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

 WITH S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 485 437 451 458 ± 25

 solvent control 28 44 33 35 ± 8 

 3 42 42 40 41 ± 1 

 10 40 39 38 39 ± 1 

 33 32 31 31 31 ± 1 

 100 30 25 32 29 ± 4 

 333 29 30 30 30 ± 1 

 1000 27 31 31 30 ± 2 

 3330 37 46 38 40 ± 5 

 5000 27 32 33 31 ± 3 

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

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 22 - 

APPENDIX 3 – continued –

Experiment 2

Strain TA1535

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

 WITHOUT S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 842 798 826 822 ± 22

 solvent control 13 11 10 11 ± 2 

 100 8 3 11 7 ± 4 

 333 8 9 12 10 ± 2 

 1000 12 8 9 10 ± 2 

 3330 8 9 13 10 ± 3 

 5000 10 7 14 10 ± 4 

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

 WITH S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 160 202 187 183 ± 21

 solvent control 8 10 8 9 ± 1 

 100 4 6 10 7 ± 3 

 333 10 6 4 7 ± 3 

 1000 9 9 4 7 ± 3 

 3330 6 7 5 6 ± 1 

 5000 11 9 9 10 ± 1 

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

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1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 23 - 

APPENDIX 3 – continued –

Experiment 2

Strain TA1537

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

 WITHOUT S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 596 595 603 598 ± 4 

 solvent control 8 4 6 6 ± 2 

 100 6 3 5 5 ± 2 

 333 8 5 3 5 ± 3 

 1000 4 4 3 4 ± 1 

 3330 3 4 5 4 ± 1 

 5000 3 5 2 3 ± 2 

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

 WITH S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 473 480 470 474 ± 5 

 solvent control 4 4 3 4 ± 1 

 100 5 5 5 5 ± 0 

 333 5 11 7 8 ± 3 

 1000 9 3 5 6 ± 3 

 3330 3 8 6 6 ± 3 

 5000 6 2 3 4 ± 2 

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

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 24 - 

APPENDIX 3 – continued –

Experiment 2

Strain TA98

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

 WITHOUT S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 1133 937 1098 1056 ± 105

 solvent control 18 13 15 15 ± 3 

 100 18 16 23 19 ± 4 

 333 19 18 21 19 ± 2 

 1000 22 22 19 21 ± 2 

 3330 22 21 20 21 ± 1 

 5000 15 23 18 19 ± 4 

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

 WITH S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 397 328 394 373 ± 39

 solvent control 21 26 34 27 ± 7 

 100 27 27 31 28 ± 2 

 333 23 21 20 21 ± 2 

 1000 23 30 30 28 ± 4 

 3330 33 24 25 27 ± 5 

 5000 26 31 27 28 ± 3 

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

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 25 - 

APPENDIX 3 – continued –

Experiment 2

Strain TA100

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

 WITHOUT S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 800 709 721 743 ± 49

 solvent control 138 126 103 122 ± 18

 100 1)

 333 130 119 127 125 ± 6 

 1000 113 174 123 137 ± 33

 3330 130 119 140 130 ± 11

 5000 117 109 131 119 ± 11

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

 WITH S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 334 346 338 339 ± 6 

 solvent control 137 96 95 109 ± 24

 100 126 126 131 128 ± 3 

 333 173 135 147 152 ± 19

 1000 129 119 149 132 ± 15

 3330 104 113 130 116 ± 13

 5000 114 135 124 124 ± 11

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

1) Due to a technical error, plates could not be determimed

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 26 - 

APPENDIX 3 – continued –

Experiment 2

Strain WP2uvrA

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

 WITHOUT S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 1023 1089 961 1024 ± 64

 solvent control 20 34 24 26 ± 7 

 100 22 18 17 19 ± 3 

 333 19 20 15 18 ± 3 

 1000 12 19 20 17 ± 4 

 3330 23 25 21 23 ± 2 

 5000 16 18 12 15 ± 3 

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

 WITH S9-MIX

 plate 1 2 3 MEAN SD

 dose (μg/plate)

 positive control 346 281 322 316 ± 33

 solvent control 24 21 34 26 ± 7 

 100 22 28 30 27 ± 4 

 333 23 31 28 27 ± 4 

 1000 34 27 37 33 ± 5 

 3330 34 31 32 32 ± 2 

 5000 15 17 20 17 ± 3 

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

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

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APPENDIX 4 HISTORICAL CONTROL DATA OF THE SOLVENT CONTROL

TA1535 TA1537 TA98 TA100 WP2uvrA

S9-mix - + - + - + - + - + 

Range 2 - 25 2 – 31 2 - 19 2 - 32 7 - 49 8 - 59 61 - 191 58 - 205 7 - 51 7 - 62

Mean 10 8 5 5 20 24 108 94 22 24 

SD 4 3 2 2 6 7 24 24 7 7 

n 1468 1469 1253 1251 1453 1451 1485 1449 1309 1290

SD = Standard deviation

n = Number of observations

Historical control data from experiments performed between January 2010 and January 2013. 

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 28 - 

APPENDIX 5 HISTORICAL CONTROL DATA OF THE POSITIVE CONTROL SUBSTANCES

TA1535 TA1537 TA98

S9-mix - + - + - + 

Range 245 - 1270 72 – 1016 113 - 1182 85 - 677 407 - 1518 202 - 1663 

Mean 841 259 539 353 1008 806

SD 122 116 265 103 134 290

n 1431 1437 1237 1242 1446 1428

TA100 WP2uvrA

S9-mix - + - + 

Range 348 - 1417 229 - 1752 138 - 1479 122 - 1233

Mean 872 1114 1058 358

SD 130 238 209 125

n 1452 152 1280 1264

SD = Standard deviation

n = Number of observations

Historical control data from experiments performed between January 2010 and January 2013. 

Final Report
1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) Project 500680

- Page 29 - 

APPENDIX 6 CERTIFICATE OF ANALYSIS

Final Report