Formamide, N-[(2-bromo-5-hydroxy-4-methoxyphenyl)methyl]-N-[2-(4-hydroxyphenyl)ethyl]-

Hydrolysis

Link to relevant study record(s)

Reference

Endpoint:

hydrolysis

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2017-04-20 to 2017-06-26

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:

yes

Remarks:

See section: Any other information on materials and methods incl. tables

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

See section: Any other information on materials and methods incl. tables

Qualifier:

according to guideline

Guideline:

EPA OPPTS 835.2120 (Hydrolysis of Parent and Degradates as a Function of pH at 25°C)

Deviations:

yes

Remarks:

See section: Any other information on materials and methods incl. tables

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: I15EB1917
- Expiration date of the lot/batch: 2017-09-07 (retest date)
- Purity: 98.9% (based on HPLC)
- Purity test date: 2017-06-07 (certificate of analysis release date)

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: not indicated

Radiolabelling:

no

Analytical monitoring:

yes

Details on sampling:

- Sampling intervals for the parent/transformation products:
* Tier 1: at the beginning and after 5 days
* Tier 2: at the beginning and at several sampling points after t=0

- Sampling method:
* Tier 1: Concentration of the test item in the test sample was determined immediately after preparation. The samples taken at t=5 days were cooled to room temperature using running tap water. The samples were diluted in a 4:1 (v:v) ratio with acetonitrile and analyzed.
* Tier 2: Test samples were prepared and treated similarly as during the preliminary test.

Buffers:

- pH:
* Tier 1: pH 4.0, pH 7.0 and pH 9.0
- Tier 2: pH 7.0 and pH 9.0
- Type and final molarity of buffer:
* Acetate buffer pH 4, 0.1 M: Solution of 16.7% 0.1 M sodium acetate in water and 83.3% 0.1 M acetic acid in water. Buffer contained 0.0009% (w/v) sodium azide.
* Phosphate buffer pH 7, 0.1 M: Solution of 0.1 M potassium di-hydrogen-phosphate in water adjusted to pH 7 using 10N sodium hydroxide. Buffer contained 0.0009% (w/v) sodium azide.
* Borate buffer pH 9, 0.1 M: Solution of 0.1 M boric acid in water and 0.1 M potassium chloride in water adjusted to pH 9 using 10N sodium hydroxide. Buffer contained 0.0009% (w/v) sodium azide.

Details on test conditions:

TEST SYSTEM
- Type, material and volume of test flasks, other equipment used: sterile vessel
- Sterilisation method: 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.
- Lighting: in the dark
- Measures taken to avoid photolytic effects: vessels placed in the dark
- Measures to exclude oxygen: Nitrogen was passed through the buffer solutions for about 5 minutes.
- Temperature: 50.1°C +/- 0.1°C.
- If no traps were used, is the test system closed/open: under vacuum
- Is there any indication of the test material adsorbing to the walls of the test apparatus? no

TEST MEDIUM
- Volume used/treatment: 6 mL
- Kind and purity of water: Tap water purified by a Milli-Q water purification system (Millipore, Bedford, MA, USA)
- Preparation of test medium: The test item was spiked to the solutions at a target concentration of 1.9 mg/L using a spiking solution in DMSO.
- Renewal of test solution: not applicable
- Identity and concentration of co-solvent: not applicable

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Dissolved oxygen: no data

Duration:

5 d

pH:

4

Temp.:

50 °C

Initial conc. measured:

>= 0.971 - <= 0.977 mg/L

Duration:

5 d

pH:

7

Temp.:

50 °C

Initial conc. measured:

>= 0.998 - <= 1 mg/L

Duration:

5 d

pH:

9

Temp.:

50 °C

Initial conc. measured:

>= 0.993 - <= 0.999 mg/L

Duration:

121.03 h

pH:

7

Temp.:

50 °C

Initial conc. measured:

>= 1.07 - <= 1.07 mg/L

Duration:

117.97 h

pH:

7

Temp.:

70 °C

Initial conc. measured:

>= 1.04 - <= 1.07 mg/L

Duration:

439.23 h

pH:

9

Temp.:

50 °C

Initial conc. measured:

>= 1.01 - <= 1.03 mg/L

Duration:

437.63 h

pH:

9

Temp.:

70 °C

Initial conc. measured:

>= 1.02 - <= 1.05 mg/L

Number of replicates:

- Calibration solutions were injected in duplicate.
- Test samples were analyzed by single injection.

Positive controls:

no

Negative controls:

no

Statistical methods:

No data

Preliminary study:

The preliminary hydrolysis tests was performed at 50.1°C ± 1°C. At pH 7 and pH 9, a decrease in concentration of ≥10% was observed after 5 days. According to the guideline, the higher Tier test was required to determine the half-life time of the test item.
A degree of hydrolysis of < 10% was observed at pH 4 after 5 days. It demonstrated that the half-life time of the test item at 25°C is > 1 year. According to the guideline, no further tests were required.
Small responses at the retention time of the test item were observed in chromatograms of some of the blank buffer solutions. Contribution to the test samples based on peak area was <1% and therefore considered negligible.
The mean recoveries of the of the test item containing buffer solutions at t=0 fell within the criterion range of 90-110%. It demonstrated that the analytical method was adequate to support the hydrolysis study on the test item.

Transformation products:

no

% Recovery:

97

pH:

4

Temp.:

50 °C

Duration:

5 d

Remarks on result:

hydrolytically stable based on preliminary test

% Recovery:

100

pH:

7

Temp.:

50 °C

Duration:

5 d

Remarks on result:

hydrolytically stable based on preliminary test

% Recovery:

100

pH:

9

Temp.:

50 °C

Duration:

5 d

Remarks on result:

hydrolytically stable based on preliminary test

% Recovery:

107

pH:

7

Temp.:

50 °C

Duration:

121.03 h

% Recovery:

106

pH:

7

Temp.:

70 °C

Duration:

117.97 h

% Recovery:

102

pH:

9

Temp.:

50 °C

Duration:

439.23 h

% Recovery:

104

pH:

9

Temp.:

70 °C

Duration:

437.63 h

pH:

4

Temp.:

25 °C

DT50:

> 1 yr

Type:

(pseudo-)first order (= half-life)

Key result

pH:

7

Temp.:

25 °C

DT50:

> 1 yr

Type:

(pseudo-)first order (= half-life)

pH:

9

Temp.:

25 °C

DT50:

> 1 yr

Type:

(pseudo-)first order (= half-life)

Details on results:

- Small responses at the retention time of the test item were observed in chromatograms of some of the blank buffer solutions. Contribution to the test samples based on peak area was <1% and therefore considered negligible.
- The mean recoveries of the test item containing buffer solutions at t=0 fell within the criterion range of 90-110%.

Validity criteria fulfilled:

yes

Conclusions:

The preliminary test (Tier 1) and main study (Tier 2) according to OECD 111 were performed for the determination of the rate of hydrolysis of JNJ-28645409-AAA (T002078) at pH values normally found in the environment (pH 4-9) . The half life at pH 4, 7 and 9 at temperature 25°C was found to be > 1 year. Results of this study can be considered reliable without restrictions.

Description of key information

One study (Brekelmans, 2017) was performed according to OECD guideline 111 and regarded as a key study (Klimisch score of 1). A half-life time of > 1 year at 25 °C and pH 7 was determined for T002078.

Additional information

The determination of the hydrolysis as a function of pH of the substace was performed according to OECD guideline 111 (Brekelmans, 2017). The preliminary hydrolysis tests was performed at 50.1°C ± 1°C. At pH 7 and pH 9, a decrease in concentration of ≥10% was observed after 5 days. According to the guideline, the higher Tier test was required to determine the half-life time of the test item. A half-life time of > 1 year at 25 °C and pH 7 was determined for T002078.