Registration Dossier

Administrative data

Endpoint:
hydrolysis
Type of information:
experimental study
Adequacy of study:
key study
Study period:
11 February 2011 – 27 April 2011
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: This study has been performed according to OECD and EC guidelines and in compliance with GLP principles.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2011
Report Date:
2011

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 111 (Hydrolysis as a Function of pH)
Version / remarks:
(2004)
Deviations:
no
Qualifier:
according to
Guideline:
EU Method C.7 (Degradation: Abiotic Degradation: Hydrolysis as a Function of pH)
Version / remarks:
(Official Journal of the European Union no. L142, May 31, 2008)
Deviations:
no
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): E-C104
- Description: Blue powder
- Batch: MB-1
- Test substance storage: At room temperature protected from light
- Stability under storage conditions: Stable
- Expiry date: 23 December 2015
Radiolabelling:
no

Study design

Analytical monitoring:
yes
Details on sampling:
The concentration of the test substance in the test samples was determined immediately after preparation (t=0) and after 5 days. The samples taken at t=5 days were cooled to room temperature using running tap water. The samples were diluted by a factor of 2.5 with buffer pH 4, 7 or 9 and analysed.
Buffers:
- pH: 4
- Type and final molarity of buffer: Acetate buffer, 0.1 M
- Composition of buffer: solution of 16.6% 0.1 M sodium acetate and 83.4% 0.1 M acetic acid. The buffer contains 0.0009% (w/v) sodium azide.

- pH: 7
- Type and final molarity of buffer: Phosphate buffer, 0.1 M
- Composition of buffer: solution of 0.1 M potassium dihydrogenphosphate adjusted to pH 7 using 10 N sodium hydroxide. The buffer contains
0.0009% (w/v) sodium azide.

- pH: 9
- Type and final molarity of buffer: Borate buffer, 0.1 M
- Composition of buffer: solution of 0.1 M boric acid and 0.1 M potassium chloride adjusted to pH 9 using 10 N sodium hydroxide. The buffer
contains 0.0009% (w/v) sodium azide.
Details on test conditions:
Preliminary test - Tier 1:

Test substance solutions were prepared in the buffer solutions at a target concentration of 1000 mg/l. Each solution was filter-sterilised through a 0.2 μm FP 30/0.2 CA-S filter (Whatman, Dassel, Germany) and transferred into a sterile vessel. To exclude oxygen, nitrogen gas was purged through the solution for 5 minutes. For each sampling time, duplicate sterile vessels under vacuum were filled with 6 ml test solution and placed in the dark in a temperature controlled environment at 49.9°C +/- 0.2°C.

Blank buffer solutions were treated similarly as the test samples and analysed at t=0.

The pH of each of the test solutions (except for the blanks) was determined at each sampling time.
Duration of test
Duration:
5 d
Initial conc. measured:
952 - 1 017 mg/L
Number of replicates:
2 measurements at each pH (separate vessels)

Results and discussion

Preliminary study:
At pH 4, pH 7 and pH 9, a degree of hydrolysis of < 10% was observed after 5 days. It demonstrated that the half-life time of the test substance at 25°C is > 1 year. According to the guideline, no further tests were required.

No test substance was detected in blank buffer solutions.
Test performance:
The mean recoveries of the buffer solutions fell within the criterion range of 90-110%. It demonstrated that the analytical method was adequate to support the hydrolysis study on the test substance.
Total recovery of test substance (in %)open allclose all
% Recovery:
97 - 98
pH:
4
Temp.:
49.9 °C
Duration:
5 d
% Recovery:
99 - 101
pH:
7
Temp.:
49.9 °C
Duration:
5 d
% Recovery:
98 - 99
pH:
9
Temp.:
49.9 °C
Duration:
5 d

Any other information on results incl. tables

Preliminary test – hydrolysis of the test substance at pH 4, pH 7 and pH 9 measured at 268 nm

 

pH code

Sampling time

Analysed concentration
[mg/l]

Degree of hydrolysis
[%]

Actual pH

Individual

Mean

 

 

 

 

 

 

pH 4

0 hours

955

 

 

4.0

 

 

967

 

 

4.0

 

5 days

955

0.6

0.1

4.0

 

 

966

-0.5

 

4.0

 

 

 

 

 

 

pH 7

0 hours

1013

 

 

7.1

 

 

1017

 

 

7.1

 

5 days

1033

-1.8

-1.2

7.1

 

 

1021

-0.6

 

7.1

 

 

 

 

 

 

pH 9

0 hours

1015

 

 

9.0

 

 

1012

 

 

9.0

 

5 days

1027

-1.3

-1.1

9.0

 

 

1023

-0.9

 

9.0

 

 

 

 

 

 

 

Preliminary test – hydrolysis of the test substance at pH 4, pH 7 and pH 9 measured at 617 nm

pH code

Sampling time

Analysed concentration
[mg/l]

Degree of hydrolysis
[%]

Actual pH

Individual

Mean

 

 

 

 

 

 

pH 4

0 hours

952

 

 

4.0

 

 

970

 

 

4.0

 

5 days

954

0.7

0.7

4.0

 

 

954

0.7

 

4.0

 

 

 

 

 

 

pH 7

0 hours

1000

 

 

7.1

 

 

1011

 

 

7.1

 

5 days

1014

-0.9

-0.6

7.1

 

 

1010

-0.4

 

7.1

 

 

 

 

 

 

pH 9

0 hours

1010

 

 

9.0

 

 

1007

 

 

9.0

 

5 days

1004

0.5

0.5

9.0

 

 

1003

0.6

 

9.0

 

 

 

 

 

 

Applicant's summary and conclusion

Validity criteria fulfilled:
yes
Conclusions:
The preliminary test (Tier 1) was performed for the determination of the rate of hydrolysis of E-C104 at pH values normally found in the environment (pH 4-9).

At each pH value a degree of hydrolysis of < 10% was observed after 5 days. According to the guideline, performance of the main study (Tier 2) was not required. The half-life time of the test substance at 25°C was > 1 year at each pH tested.