Reaction products of Hydrogen amino-4-hydroxynaphthalene-2-sulfonate, Potassium substituted-5-{[2-(sulfonatooxy)ethyl]sulfonyl}benzenesulfonate and Sodium substituted-({4-[(2-chloroethyl)sulfonyl]butanoyl}amino)benzenesulfonate

Hydrolysis

Link to relevant study record(s)

Reference

Endpoint:

hydrolysis

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 111 (Hydrolysis as a Function of pH)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Name: FAT 40875/A TE
Physical appearance: Dark red powder.
Purity: 86.4% all organic components
Batch No: BOP 05-17 (BS-ROE 1550 NIM 25+26+27)
Manufactured Date: December, 11th 2017
Expirty date: December, 11th 2022.
Recommended Storage Condition: Refrigeration (+2 to +8 °C), Protect from light.

Radiolabelling:

no

Analytical monitoring:

yes

Details on sampling:

- Sampling intervals for the parent/transformation products: Day 0 and Day 5.
- Sampling method: HPLC
- Sampling intervals/times for pH measurements: Day 0 and Day 5.
- Sampling intervals/times for sterility check: Start date and study termination dates.
- Sample storage conditions before analysis: Samples were analysed on the sampling day.

Buffers:

- pH: 4
- Type and final molarity of buffer: Acetic acid solution, 0.2 M + Anhydrous sodium acetate solution 0.2 M.
- Composition of buffer: Acetic acid solution 205 mL + Anhydrous sodium acetate solution 45 mL.

Details on test conditions:

TEST SYSTEM
- Type, material and volume of test flasks, other equipment used: 20 mL amber coloured vials.
- Sterilisation method: All glassware used was sterilised using autoclaved before use. All buffer solutions were sterilised by passing through 0.2 µm sterilised filter. The experiment was performed in laminar flow chamber under aseptic condition. Sterility of the samples was checked at experimental start date and at study termination for each of the test systems.
- Lighting: No lighting.
- Measures taken to avoid photolytic effects: The prepared test item solutions taken in amber coloured vials were kept in a dark incubator in order to avoid photolytic effects.
- Measures to exclude oxygen: The buffer solutions were bubbled with nitrogen gas for approximately 5 minutes.


OTHER TEST CONDITIONS
- Adjustment of pH: The pH of the buffered test item solutions during the reactions were measured after equilibrating the test samples at room temperature using a calibrated pH meter. The pH of the test item solutions was measured on Day 1 and Day 5.

Duration:

5 d

pH:

4

Temp.:

50 °C

Initial conc. measured:

100 mg/L

Number of replicates:

2

Positive controls:

no

Negative controls:

yes

Remarks:

buffer solution

Preliminary study:

The hydrolysis of test item at 50 ± 0.5 °C after 5 days was found to be less than 10 % at pH 4.0. The test item was considered to be hydrolytically stable in pH 4.0 buffer (t1/2>1 Year).

Transformation products:

not specified

% Recovery:

88.1

St. dev.:

6.2

pH:

4

Temp.:

50 °C

Duration:

5 d

Remarks on result:

hydrolytically stable based on preliminary test

pH:

4

Temp.:

50 °C

DT50:

> 1 yr

Type:

not specified

Remarks on result:

hydrolytically stable based on preliminary test

Details on results:

TEST CONDITIONS
- pH, sterility, temperature, and other experimental conditions maintained throughout the study: Yes
- Anomalies or problems encountered (if yes): no

Limit of Detection (LOD) of the method was 0.77 µg/mL

Limit of Quantitation (LOQ) of the method was 5 µg/mL

Validity criteria fulfilled:

yes

Conclusions:

The test item was considered to be hydrolytically stable in pH 4.0 buffer (t1/2 >1 Year).

Executive summary:

A study was performed as per OECD Guideline No. 111 to determine the rate of hydrolysis of the test item as a function of pH at pH 4 buffer and temperature.

Test item at nominal concentration of about 100 mg/L (100 µg/mL) in sterile buffer solutions of pH 4.0 was incubated for 5 days at 50 ± 0.5 °C in the preliminary test. Hydrolysis reactions were monitored by analyzing the test item concentration at set intervals using an in-house developed and validated HPLC method.

 

The hydrolysis of test item after 5 days of incubation at 50 ± 0.5 °C was 6.8 % at pH 4.0. The hydrolysis of test item at 50 ± 0.5 °C after 5 days was found to be less than 10 % at pH 4.0. Hence, the test item was considered to be hydrolytically stable in pH 4.0 buffer (t1/2 >1 Year).

Description of key information

FAT 40875/A was considered to be hydrolytically stable in pH 4.0 buffer (t1/2 >1  Year).

Key value for chemical safety assessment

Half-life for hydrolysis:

1 yr

at the temperature of:

25 °C

Additional information

A study was performed as per OECD Guideline No. 111 to determine the rate of hydrolysis of the test item as a function of pH at pH 4 buffer and temperature. Test item at nominal concentration of about 100 mg/L (100 µg/mL) in sterile buffer solutions of pH 4.0 was incubated for 5 days at 50 ± 0.5 °C in the preliminary test.

 

The hydrolysis of test item after 5 days of incubation at 50 ± 0.5 °C was 6.8 % at pH 4.0. The hydrolysis of test item at 50 ± 0.5 °C after 5 days was found to be less than 10 % at pH 4.0. Hence, the test item was considered to be hydrolytically stable in pH 4.0 buffer (t1/2 >1 Year).