Hydrocotyle asiatica, ext.

Hydrolysis

Administrative data

Endpoint:

hydrolysis

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 111 (Hydrolysis as a Function of pH)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method C.7 (Degradation: Abiotic Degradation: Hydrolysis as a Function of pH)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Hydrocotyle asiatica, ext.

EC Number:

283-640-5

EC Name:

Hydrocotyle asiatica, ext.

Cas Number:

84696-21-9

Molecular formula:

not applicable

IUPAC Name:

Hydrocotyle asiatica, ext.

Test material form:

solid

Specific details on test material used for the study:

Test Item name: Centella asiatica dry extract
Batch No. 972/43/A
Appearance dark green, powder
Retesting date: August 31, 2020
Storage: room temperature

Study design

Buffers:

Buffer solutions
pH 4.0: 1 mL 0.2 M Sodium hydroxide and 125 mL 0.2 M Potassium hydrogen phthalate were diluted to 500 mL with Ultrapure water.
pH 7.0: 74 mL 0.2 M Sodium hydroxide and 125 mL 0.2 M Potassium dihydrogen phosphate were diluted to 500 mL with Ultrapure water.
pH 9.0: 53.5 mL 0.2 M Sodium hydroxide and 125 mL 0.2 M Boric acid and Potassium chloride were diluted to 500 mL with Ultrapure water.
These sterile buffer solutions were prepared using reagent grade chemicals and ultrapure, sterile water. The pH of each buffer solution was checked with a calibrated pH meter to a precision of at least 0.01.

Details on test conditions:

Hydrolysis was examined at three different pH values: 4.0, 7.0 and 9.0, at 50  0.5C using a dark thermostat to avoid photolytic effects

Test Solutions

About 20 mg of test item was weighted to the nearest 0.01 mg into a volumetric flask and dissolved in each buffer solution to a total volume of 200 mL.
Test solution was transferred into 20 mL headspace–vials under sterile circumstances, under a laminar flow hood. The tubes were entirely filled with the solution. From each test solution five tubes were prepared and from each control solution two tubes were prepared.
The tubes were thermostated at 50 ± 0.5 °C.

The test solutions were analysed at the start of the test and after five days with five replicate samples each.
Two replicate tubes filled with control buffers were analysed at the start and at the end of the incubation.

Aqueous and control samples were hundredfold diluted with the mobile phase and analysed by LC/MS/MS method.

Results and discussion

Transformation products:

not specified

Dissipation DT50 of parent compound

Dissipation DT50 of parent compound 1

pH:

4

Temp.:

50 °C

Remarks on result:

other: unstable

Dissipation DT50 of parent compound 2

pH:

7

Temp.:

50 °C

Remarks on result:

other: unstalbe

Dissipation DT50 of parent compound 3

pH:

9

Temp.:

50 °C

Remarks on result:

other: unstable

Applicant's summary and conclusion

Conclusions:

The hydrolysis test was performed at 50  0.5 C, at pH 4, 7 and 9.
In the course of the preliminary test Madecassicoside and Asiaticoside components was found to be unstable at pH 4, 7 and 9 at 50 ± 0.5 °C, therefore the hydrolysis main test will be needed to perform.