Registration Dossier

Administrative data

Endpoint:
hydrolysis
Type of information:
experimental study
Adequacy of study:
key study
Study period:
Start of experiment: 02 December 2009 End of experiment: 08 December 2009
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies which did not affect the quality of relevant results.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2010
Report date:
2010

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 111 (Hydrolysis as a Function of pH)
Qualifier:
according to guideline
Guideline:
EU Method C.7 (Degradation: Abiotic Degradation: Hydrolysis as a Function of pH)
GLP compliance:
yes (incl. QA statement)
Remarks:
Date of inspection: 13th to the 22nd of October 2008 (GLP certificate is valid for 2 years)

Test material

Constituent 1
Reference substance name:
Pellicer
IUPAC Name:
Pellicer
Details on test material:
Name: Pellicer
Lot No.: 01-09-17
Chemical name: L-Lysine,N2,N6-bis[N-(1-oxododecyl)-L-α(or γ)-glutamyl]-, sodium salt (1;?)
Appearance: White powder
Storage condition: Room temperature (15-25 °C), in the dark and protected from humidity (with desiccant)
Manufacture date: 17 January 2009
Expiry Date: 17 January 2011
Safety precautions: Routine safety and hygienic procedures sufficient to ensure personnel health and safety

Study design

Analytical monitoring:
yes
Details on sampling:
- Sampling method: The reaction solutions were analysed at the start of the test and after five days with five replicate samples each.
- Sampling intervals/times for pH measurements: The pH of each buffer solution was checked with a calibrated pH meter.
- Sample storage conditions before analysis: The tubes were thermostated at 50 °C ± 1.0 °C. : Solutions were transferred into 25 ml screw cap tubes. Five tubes were prepared from each test solution
Buffers:
Buffer solutions: : Hydrolysis was examined at three different pH values: 4.0, 7.0 and 9.0 in the dark.

pH 4.0: 1 ml 0.2 M Sodium hydroxide and 125 ml 0.2 M Potassium hydrogen phthalate were be diluted to 500 ml with ultra-pure 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 ultra-pure 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 ultra-pure water. These sterile buffer solutions were prepared using reagent grade chemicals and ultra-pure, sterile water.

The pH of each buffer solution was checked with a calibrated pH meter.
Details on test conditions:
TEST SYSTEM
- Type, material and volume of test flasks, other equipment used:
- Sterilisation method: Nitrogen was bubbled into the water for five minutes before the preparation of the solutions in order to exclude oxygen.
- Lighting: carried out in the dark
- Measures taken to avoid photolytic effects: The hydrolysis reaction was carried out using a dark thermostat to avoid photolytic effects.
- Measures to exclude oxygen: Nitrogen was bubbled into the water for five minutes before the preparation of the solutions in order to exclude oxygen.
- If no traps were used, is the test system closed
- Dissolved oxygen: Nitrogen was bubbled into the water for five minutes before the preparation of the solutions in order to exclude oxygen.
Duration of testopen allclose all
Duration:
5 d
pH:
4
Temp.:
50
Initial conc. measured:
>= 5 - <= 100 other: µg/ml
Duration:
5 d
pH:
7
Temp.:
50
Initial conc. measured:
>= 5 - <= 100 other: µg/ml
Duration:
5 d
pH:
9
Temp.:
50
Initial conc. measured:
>= 5 - <= 100 other: µg/ml
Number of replicates:
5
Positive controls:
not specified
Negative controls:
not specified
Statistical methods:
None stated in the report

Results and discussion

Preliminary study:
The hydrolysis test was performed at 50 ± 1.0 °C, at pH 4, 7 and 9. Measured concentrations and pH values are summarised in Table 3. Please see this in the attachements section. In the course of the preliminary test Pellicer was proved to be hydrolytically stable at pH 4, 7 and 9.
Test performance:
The calibration series was prepared in water. It was measured at both analytical occasions. Concentrations of the calibration samples were 5, 10, 25, 50, and 100 μg / ml.
Transformation products:
no
Total recovery of test substance (in %)open allclose all
% Recovery:
< 10
pH:
4
Temp.:
50 °C
% Recovery:
< 10
pH:
7
Temp.:
50 °C
% Recovery:
< 10
pH:
9
Temp.:
50 °C
Details on results:
Pellicer is considered to be hydrolytically stable and therefore no additional testing is required.

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Remarks:
The validity criteris was fulfilled for this study
Conclusions:
The observed hydrolysis of Pellicer was less than 10 per cent after 5 days at a temperature of 50 °C at each of the three pH values.
Executive summary:

In the course of the preliminary test Pellicer proved to be hydrolytically stable at pH 4, 7 and 9. The observed hydrolysis of Pellicer was less than 10 per cent after 5 days at a temperature of 50 °C at each of the three pH values. Therefore Pellicer is considered to be hydrolytically stable under the conditions of this test.