Endpoint study record: Analytical methods. HPLC-RI. WIL Research Project 500674

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Printing Date 2013-06-12 12:19:03 CEST

Restriction of specific regulatory purposes

Confidentiality

Owner (TS 204240) 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) / WIL Research Europe B.V. /

's-Hertogenbosch / Netherlands

Legal entity

owner

WIL Research Europe B.V. / 's-Hertogenbosch / Netherlands

Endpoint study record: Analytical methods. HPLC-RI. WIL Research

Project 500674

UUID IUC5-e39aad31-4419-4cd5-a046-7ea12223d61e

Dossier UUID 0

Author TRJ / WIL Research Europe B.V. / 's-Hertogenbosch / Netherlands

Date 2013-06-12 12:18:54 CEST

Remarks

Administrative Data

Purpose flag key study; robust study summary

Study result type experimental result Study period 07 March 2013 - 03 May 2013

Reliability 1 (reliable without restriction)

Rationale for reliability incl.

deficiencies

The study has been performed according to EC recommendations and in compliance with the

principles of GLP.

Data source

Background information

A high performance liquid chromatographic method with refractive index (RI) detection was validated for the quantitative analysis of the

substance in water. The validation included the determination of the following parameters:

- Specificity

- Linearity

- Accuracy and repeatability

- Limit of quantification

- Stability of the analytical system and end solutions

- Stability of stock solutions

- Storage stability of samples

Reference
Endpoint study record: Analytical methods. HPLC-RI. WIL Research Project 500674

file:///C|/...net Files/Content.Outlook/6RSURWTF/Endpoint study record Analytical methods_ HPLC-RI_ WIL Research Project 500674.html[9/13/2013 2:27:56 PM]

Reference

type

Author Year Title Bibliographic

source

Testing

laboratory

Report

no.

Owner

company

Company

study no.

Report

date

study

report

Brekelmans,

MJC

2013 Development and

validation of an

analytical method for

the analysis of 1-

Propanesulfonic acid,

2-hydroxy-3-(2-

propen-1-yloxy)-,

sodium salt (1:1) in

water

WIL Research

Europe B.V.,

Hambakenwetering

7, 5231 DD ‘sHertogenbosch,

The Netherlands

500674 Society of

Chemical

Manufacturers

and Affiliates,

1850 M Street,

NW Suite 700,

WASHINGTON,

DC 20036, USA

2013-

06-10

Data protection claimed

yes

Materials and methods

Matrix / medium

water

Test guideline

Qualifier Guideline Deviations

according

to

other guideline: 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).

no

GLP compliance

yes

Test materials

Identity of test material same as for substance defined in section 1 (if not read-across)

yes

Test material identity

Identifier Identity

CAS number 52556-42-0

Test material form

other: liquid

Details on test material

- Name of test material (as cited in study report): 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1)

- Description: Clear colourless liquid 

- Storage condition of test material: At room temperature in the dark

Confidential details on test material

- Purity/Composition: 38.64% of CAS# 52556-42-0, 2.26% of CAS# 123-34-2, 59.1% of water 

- Lot/batch No.: 12E1101

- Expiration date of the lot/batch: 13 August 2013

Principles of analytical methods

Instrument / detector

other: HPLC-RI

Details on analytical method

Instrument: Alliance Separation Module 2695 (Waters, USA) 

Detector: Refractive Index Detector 2414 (Waters) 

Column: Synergi Hydro-RP 80A, 250 mm x 4.6 mm i.d., dp =4 μm (Phenomenex, USA)

Column temperature: 40°C ± 1°C 

Injection volume: 50 μL 

Mobile phase: Water containing 0.05M Na2HPO4, 0.05M NaH2PO4 and 0.05M NaNO3 
Endpoint study record: Analytical methods. HPLC-RI. WIL Research Project 500674

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Flow: 1 mL/min

RI detection: 30°C; sensitivity of 4; time constant of 1 seconds

Stock solutions of the substance in water: 5530 - 6660 mg/L (nominal)

Five calibration solutions (80 – 1200 mg/L nominal) in mobile phase: prepared from two stock solutions

Chromatogram: one test substance peak (see attached document).

Response: peak area of the test substance. 

Confirmatory method (if applicable)

Any other information on materials and methods incl. tables

SPECIFICITY

Single injection of test substance solution and a blank accuracy sample. If a peak at the retention time of the test

substance was present in the blank chromatogram, the response was compared with the limit of quantification.

LINEARITY

Duplicate analysis of five calibration solutions.The responses were plotted against the nominal concentrations using regression analysis

(linear least squares method) with a 1/concentration^2 weighting factor.

ACCURACY AND REPEATABILITY

Matrix: tap water purified by a water purification system (Millipore, USA)

Spiking solutions in water: (prepared from) a test substance stock solution in water.

Accuracy samples: Water was spiked with test substance at a target concentration of 100 or 1000 mg/L. The accuracy samples were

analysed without further treatment. The blank accuracy sample was prepared and treated similarly to the accuracy samples. Accuracy

samples were analysed by single injection.

LIMIT OF QUANTIFICATION (LOQ)

LOQ is the lowest concentration level at which an accuracy in the range 70-110% and a repeatability of = 20% was demonstrated and

was obtained from the data of the accuracy- and repeatability test.

STABILITY OF THE ANALYTICAL SYSTEM AND END SOLUTIONS

Each of three calibration solutions was injected throughout the validation sequence including the beginning and end.

STABILITY OF STOCK SOLUTIONS

Single injection of calibration solutions prepared from two fresh and two stored stock solutions. Storage conditions: at room temperature

for at least 12 hours.

STORAGE STABILITY OF SAMPLES

Additional accuracy samples were prepared and stored in the freezer (= -15°C) for at least 16 hours. On the day of analysis, the samples

were defrosted at room temperature and treated identically as freshly analysed accuracy samples.

Results and discussions

Recovery results and characteristics of analytical method

Characteristics of analytical method

SPECIFICITY

The chromatogram of the test substance solution showed one test substance peak. The chromatogram of the blank accuracy sample

showed no peak at the retention time of the test substance. Therefore the analytical method was found to be specific for the substance. 

LINEARITY

There was a linear relationship between response and test substance concentration in the range of 79.9 – 1200 mg/L. Since the

coefficient of correlation (r) was > 0.99 and the back calculated accuracies were in the range 85-115% the regression line was accepted.

Regression line: see attached document.

- Slope: 6.80E01

- Intercept: -1.55E02

- Weighting factor: 1/concentration^2

- r: 0.99993

ACCURACY AND REPEATABILITY

The mean accuracy at each concentration level fell in the criterion range of 70-110% and the coefficient of variation was = 20% (see
Endpoint study record: Analytical methods. HPLC-RI. WIL Research Project 500674

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table below). The analytical method was therefore accepted for the analysis of the test substance in water in the target concentration

range of 100 - 1000 mg/L. 

LIMIT OF QUANTIFICATION

The LOQ was assessed at 100 mg/L in water. 

STABILITY OF THE ANALYTICAL SYSTEM AND END SOLUTIONS

- CV (responses) over a 19.5-h time interval at 79.9 mg/L nominal: 1.7% (n=8)

- CV (responses) over a 19.5-h time interval at 156 mg/L nominal: 1.5% (n=8)

- CV (responses) over a 18.1-h time interval at 1200 mg/L nominal: 1.3% (n=8)

Since the CV values were = 20% the analytical system and end solutions were found stable over at least a 18.1-h time interval.

STABILITY OF THE STOCK SOLUTIONS

Because the CV (response factors) of the calibration solutions prepared with fresh and stored stock solutions was = 10% (2.1% was

measured) the stock solutions were stable when stored at room temperature for at least 1 day.

STORAGE STABILITY OF SAMPLES

Mean accuracy of the 100 mg/L accuracy samples stored in the freezer was 111% which is outside the criterion range of 70 – 110%.

Since this is only slightly outside the range and since mean accuracy of the stored 1000 mg/L accuracy samples was 102%, it was still

concluded that the samples were stable during storage in the freezer.

Independent laboratory validation (if applicable)

Any other information on results incl. tables

Accuracy and Repeatability

Target

concentration

[mg/l]

Nominal

concentration

[mg/l]

Analysed

concentration

[mg/l]

Accuracy

[%]

Mean accuracy

[%]

Coefficient of

variation

[%]

100 102 110 108 106 1.8

102 109 107

102 109 107

102 106 104

102 106 103

1000 1020 1039 102 104 1.3

1020 1073 105

1020 1072 105

1020 1071 105

1020 1060 104

Storage stability of samples in the freezer

Target concentration

[mg/l]

Nominal concentration

[mg/l]

Analysed concentration

[mg/l]

Accuracy

[%]

Mean accuracy

[%]

100 104 121 117 111

104 110 106

1000 1035 1050 101 102

1035 1056 102

Overall remarks, attachments

Attached background material

Attached document Remarks

HPLC chromatograms and Regression line-TS 204240.pdf /

23.61 KB (application/octet-stream)

HPLC-RI chromatogram of 156 mg/L calibration solution and of blank

accuracy sample + Regression line

Applicant's summary and conclusion

Conclusions
Endpoint study record: Analytical methods. HPLC-RI. WIL Research Project 500674

file:///C|/...net Files/Content.Outlook/6RSURWTF/Endpoint study record Analytical methods_ HPLC-RI_ WIL Research Project 500674.html[9/13/2013 2:27:56 PM]

For the quantitative analysis of 1-Propanesulfonic acid, 2-hydroxy-3-(2-propen-1-yloxy)-, sodium salt (1:1) in water, an HPLC-RI method

was developed and successfully validated for the following parameters:

Specificity: specific

Linearity (79.9 – 1200 mg/L, nominal): r = 0.99993

Accuracy (100 and 1000 mg/L, nominal): 106 and 104%

Repeatability (100 and 1000 mg/L, nominal): 1.8 and 1.3%

Limit of quantification: 100 mg/L

Stability analytical system and end solutions (room temp. 18.1 h): stable

Stability stock solutions (room temp., 1 day): stable

Storage stability (= -15°C, at least 16 h): stable

The study has been performed according to EC recommendations and in compliance with the principles of GLP.