FINAL REPORT

Study Title

DEVELOPMENT AND VALIDATION OF 

AN ANALYTICAL METHOD FOR THE ANALYSIS OF 

CASTOR OIL, SULFATED, SODIUM SALT, 75% IN WATER

Author

I. van Doormalen - Schaap, BSc.

Test Facility

WIL Research Europe B.V.

Hambakenwetering 7

5231 DD ‘s-Hertogenbosch

The Netherlands

 

Laboratory Project Identification

Project 503334

Substance 205003/A

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Castor Oil, Sulfated, Sodium Salt, 75% Project 503334

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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. Aim of the study......................................................................................................................6

5.3. Storage and retention of records and materials .....................................................................6

6. MATERIALS......................................................................................................................................7

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

6.2. Electronic data capture...........................................................................................................7

6.3. List of deviations .....................................................................................................................7

7. VALIDATION OF AN ANALYTICAL METHOD.................................................................................8

7.1. Guidelines...............................................................................................................................8

7.2. Statement ...............................................................................................................................8

8. CARBON CONTENT ........................................................................................................................9

8.1. Reagents ................................................................................................................................9

8.2. Performance of the study .......................................................................................................9

8.3. Analytical method ...................................................................................................................9

8.4. Results................................................................................................................................. 10

8.5. Conclusion ........................................................................................................................... 10

APPENDICES

Appendix 1 Certificate of analysis ........................................................................................................ 11


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4. SUMMARY

Castor Oil, Sulfated, Sodium Salt, 75% is a complex mixture containing many sulfated and nonsulfated components. 

A general method is considered the best option for the analytical support of ecotoxicity studies with 

daphnids and algae and for the analytical support of a combined 28-day repeated dose toxicity study. 

Upon request of the sponsor, feasibility of SO3 content determination by titration was tested. The 

method which was successfully used for determination of the SO3 content in the test substance itself, 

proved not sufficiently sensitive to support ecotoxicity studies in the concentration range of 1-100 mg/l.

Total Organic Carbon (TOC) was considered to be a good alternative, since the test substance 

dissolves in aqueous solutions up to 200 mg/ml. TOC was therefore used for analysis of the samples 

from ecotoxicity studies with daphnids and algae and from the combined 28-day repeated dose toxicity 

study,

TOC analysis is a technique that does not require test substance specific validation. Therefore 

validation of an analytical method was omitted. Instead TOC content of the test substance was 

determined in order to convert concentrations from mg C/l measured to mg test substance/l. 
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5. INTRODUCTION

5.1. Preface

Sponsor Sulfated Castor Oil Consortium, Society of Chemical 

Manufacturers & Affiliates

1850 M Street NW, Suite 700

Washington, DC 20036

USA

Study Monitor Rodney Boatman

PCA Services

2704 Trail Wood Drive

Durham, NC 27705

USA

Test Facility WIL Research Europe B.V.

Hambakenwetering 7

5231 DD ‘s-Hertogenbosch

The Netherlands

Study Director I. van Doormalen - Schaap, BSc.

Study Plan Start : 15 October 2013

Completion : 03 December 2013

5.2. Aim of the study

The aim of the study was to develop and to validate an analytical method for the quantitative analysis 

of Castor Oil, Sulfated, Sodium Salt, 75% in water, ISO-medium, M2-medium and M7-medium vehicle,

to support the ecotoxicity studies with daphnids and algae and the combined 28-day repeated dose 

toxicity study.

5.3. Storage and retention of records and materials

Records and materials pertaining to the study including protocol, raw data, specimens (except 

specimens requiring refrigeration or freezing) 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.

Those specimens requiring refrigeration or freezing will be retained by WIL Research Europe for as 

long as the quality of the specimens permits evaluation but no longer than three months after 

finalization of the report.

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.
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6. MATERIALS

6.1. Test substance

6.1.1. Test substance information

Identification Castor Oil, Sulfated, Sodium Salt, 75%

CAS Number 68187-76-8

Description Clear amber liquid

Batch R-10135-064

Purity/Composition 74.4%

Test substance storage At room temperature in the dark

Stable under storage conditions until 18 July 2014 (expiry date) 

The certificate of analysis is shown in Appendix 1.

6.1.2. Study specific test substance information

Purity/composition correction factor required Yes, correction factor is 1.34 (74.4% purity)

6.2. Electronic data capture

System control, data acquisition and data processing were performed using the following programme:

Shimadzu TOC-Controle V version 2.10 (Shimadzu, Kyoto, Japan)

Temperature, relative humidity and/or atmospheric pressure during sample storage and/or 

performance of the studies was monitored continuously using the following programme:

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

USA)

6.3. List of deviations

6.3.1. List of protocol deviations

There were no deviations from the protocol.

6.3.2. List of standard operating procedures deviations

There were no deviations from standard operating procedures (SOPs).
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7. VALIDATION OF AN ANALYTICAL METHOD

7.1. Guidelines

European Commission: Guidance for Generating and Reporting Methods of Analysis in Support of 

Pre-registration Data Requirements for Annex II (Part A, section 4) and Annex III (Part A, section 5) of 

Directive 91/414, SANCO/3029/99 rev. 4 (11/07/00).

7.2. Statement

Castor Oil, Sulfated, Sodium Salt, 75% is a complex mixture containing many sulfated and nonsulfated components. 

Based on the complexity of the test substance and based on experience, development of a 

chromatographic method for analysis of all individual compounds is presumed not possible.

Gas chromatography fails since the test substance contains compounds with a relatively high 

molecular weight and of which part of it are salts. Liquid chromatography (HPLC) with 

spectrophotometric detection fails since the compounds in the test substance do not contain 

chromophoric groups. HPLC with refractive index detection is not sufficiently sensitive to measure 

concentrations in the concentration range of 1-100 mg/l i.e. the concentration range to be used in the 

ecotoxicity studies. HPLC with mass spectrometric detection fails on the differences in ionization of the 

various compounds in the test substance and on ion-suppression of co-eluting compounds. Full 

separation of this complex mixture is not possible. 

Since also spectroscopy is not an alternative since the compounds in the test substance do not 

contain chromophoric groups, a general method is considered the best alternative for the analytical 

support of ecotoxicity studies with daphnids and algae and for the analytical support of a combined 28-

day repeated dose toxicity study. Upon request of the sponsor, feasibility of SO3 content determination 

by titration, which is a general method, was tested. The method which was successfully used for 

determination of the SO3 content in the test substance itself, proved not sufficiently sensitive in the 

concentration range of 1-100 mg/l (results archived in the raw data).

Since the test substance dissolves in aqueous solutions up to 200 mg/ml, analysis of Total Organic 

Carbon (TOC) which is another general method was considered to be a good alternative for the

analysis of samples from the above mentioned studies. 

TOC analysis is a technique that does not require test substance specific validation. Therefore

validation of an analytical method was omitted. Instead TOC content of the test substance was 

determined in order to convert concentrations from mg C/l measured to mg test substance.
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8. CARBON CONTENT

8.1. Reagents

Sodium carbonate Merck, Darmstadt, Germany

Art.nr. 1.06649, Batch A0110492944.

Glucose Sigma-Aldrich, Steinheim, Germany.

Art. Nr. 67528, Batch 108k00301

All reagents were of analytical grade purity, unless specified otherwise. 

8.2. Performance of the study

For quantitative analysis of the test substance in aqueous solutions, Total Organic Carbon (TOC) 

analysis in the Non-Purgable Organic Carbon (NPOC) mode was used. In this mode, volatile 

compounds were not included.

This analytical technique does not require test substance specific validation. To calculate test

substance concentration from the measured carbon concentration, the carbon content of the test 

substance was determined.

Four samples (i.e. two samples for total carbon content and two samples for inorganic carbon content) 

of the test substance were taken for determination of the Total Organic Carbon (TOC) content. 

Calibration standards were Glucose as total carbon (TC) standard and Sodium carbonate as inorganic 

carbon (IC) standard. 

8.3. Analytical method

8.3.1. Analytical conditions

Instrument TOC-VCPH total organic carbon analyzer with SSM-5000A 

(Solid Sample module for Total Organic Carbon Analyzer) 

(Shimadzu, Kyoto, Japan)

Number of repeats 1

8.3.2. Sample measurements

The performance of the TOC analyzer was checked using Glucose and Sodium carbonate, duplicate 

samples were accurately weighed and analyzed once. The analytical system was considered suited 

for use when the carbon content of glucose (i.e. 40% carbon) was within 38 - 42% and the carbon 

content sodium carbonate (i.e. 56.6% carbon) was within 53.8 - 59.4%.

Duplicate test substance samples were accurately weighed and were analyzed once.
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8.4. Results

The carbon content results of glucose were 39.92 and 41.77%. The carbon content results of sodium 

carbonate were 53.85 and 55.38%. Since the results were within the criteria, the analytical system was

considered suited for determination of the Total Organic Carbon of the test substance. 

The results of the total carbon (TC) en inorganic carbon (IC) content of the test substance are given in 

Table 1. The Total Organic Carbon (TOC) is defined as the total carbon content minus the inorganic 

carbon content. The Total Organic Carbon is 54.48%. 

Table 1 The total and inorganic carbon content of test substance

TC content IC content

Sample 1 53.17% 0.04%

Sample 2 55.83% 0.00%

Mean 54.50% 0.02%

8.5. Conclusion

The Total Organic Carbon (TOC) content of the test substance was determined to be 54.48%. 
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Appendix 1 Certificate of analysis