Benzonitrile, 4-[(4-chloro-2-pyrimidinyl)amino]-

Hydrolysis

Link to relevant study record(s)

Reference

Endpoint:

hydrolysis

Type of information:

experimental study

Adequacy of study:

key study

Study period:

31 August 2016 - 21 November 2016

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)

Qualifier:

according to guideline

Guideline:

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

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

yes

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: I15FC2164, Janssen Pharmaceutica N.V.
- Expiration date of the lot/batch: 23-JUN-2017
- Manufacture date: 24-JUN-2015
-Certificate of analysis date: 01-JUL-2015

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature until 23-JUN-2017.

Radiolabelling:

no

Analytical monitoring:

yes

Details on sampling:

- Sampling intervals for the parent/transformation products:
* Tier 1: at the beginning and after 5 days

- 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.

Buffers:

-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 10 or 1N sodium hydroxide. Buffer contained 0.0009% (w/v) sodium azide.
-Borate buffer pH 9, 0.01 M: Solution of 0.1 M boric acid in water and 0.1 M potassium chloride in water adjusted to pH 9 using 1 or 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.:

40 °C

Initial conc. measured:

ca. 0.205 mg/L

Duration:

5 d

pH:

9

Temp.:

40 °C

Initial conc. measured:

ca. 0.202 mg/L

Duration:

5 d

pH:

4

Temp.:

40 °C

Initial conc. measured:

ca. 0.202 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:

Preliminary test - Tier 1
The buffer solutions were 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. The test item was spiked to the solutions at a target concentration of 0.200 mg/L using a spiking solution in acetonitrile. 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 50.0°C ± 0.4°C.
Note: the spiking volume was < 1% of the sample volume. Nominal concentrations were not corrected for the spiking volume.
The concentration of the test item 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 in a 1:1 (v:v) ratio with acetonitrile and analysed. Blank buffer solutions containing a similar content of blank spiking solution 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.

Transformation products:

no

% Recovery:

103 - 105

pH:

4

Temp.:

20 °C

Duration:

5 d

Remarks on result:

hydrolytically stable based on preliminary test

% Recovery:

107 - 108

pH:

7

Temp.:

50 °C

Duration:

5 d

Remarks on result:

hydrolytically stable based on preliminary test

% Recovery:

109 - 113

pH:

9

Temp.:

60 °C

Duration:

5 d

Remarks on result:

hydrolytically stable based on preliminary test

Key result

pH:

7

Temp.:

20 °C

DT50:

> 10 000 h

Type:

other: calculated according to the Gustafson and Holden Model or First-Order Multi-Compartment (FOMC) Model using Computer Aided Kinetic Evaluation (CAKE) Version 3.3, developed by Tessella Ltd, Abingdon, Oxfordshire, UK, sponsored by Syngenta.

Key result

pH:

7

Temp.:

50 °C

DT50:

565 h

Type:

other: calculated according to the Gustafson and Holden Model or First-Order Multi-Compartment (FOMC) Model using Computer Aided Kinetic Evaluation (CAKE) Version 3.3, developed by Tessella Ltd, Abingdon, Oxfordshire, UK, sponsored by Syngenta.

Key result

pH:

4

Temp.:

20 °C

DT50:

8.26 h

Type:

other: calculated according to the Gustafson and Holden Model or First-Order Multi-Compartment (FOMC) Model using Computer Aided Kinetic Evaluation (CAKE) Version 3.3, developed by Tessella Ltd, Abingdon, Oxfordshire, UK, sponsored by Syngenta.

The DT50 values of the test item are summarized in the following table:

	DT50 (95% Confidence intervals) in hours
Temperature (°C)	pH 4	pH 7	pH 9
20	>10,000	> 10,000	> 10,000
50	174 (59 to 441)	565 (0^1 to 746826)	744 (0^1 to 16039)
60	7.67 (4.64 to 12.0)	8.26 (3.71 to 15.8)	13.4 (6.7 to 23.9)

¹ Variability and trend resulted in a negative value for the lower limit of the 90% Confidence Interval. As this is not

possible, the lower limit of the 90% Confidence Interval is reported as 0.

 

Validity criteria fulfilled:

yes

Conclusions:

The determination of the hydrolysis as a function of pH of JNJ-4754724-AAA (T002488) was performed according to the OECD guidelines for the testing of chemicals no. 111: “Hydrolysis as a Function of pH", April 13, 2004. Since for pH 4, 7 and 9 more than 10% hydrolysis was observed after 5 days an additional test was performed.

At pH 4, pH 7 and pH 9 no pseudo-first order reaction kinetic was observed above room temperature. Reaction rates were not constant during the experiment. Accordingly the Arrhenius equation could not be used to determine the rate constant and half-life time at 25°C. Instead, values for disappearance time 50 (DT50) were calculated according to the Gustafson and Holden Model or First-Order Multi-Compartment (FOMC) Model using Computer Aided Kinetic Evaluation (CAKE) Version 3.3. Confidence intervals of the DT50 were back calculated using the 90% Confidence Intervals of the α and β parameter as determined with CAKE. Confidence intervals of the DT50 were not calculated if the DT50 was >10000, as variability and trend of the corresponding data did not allow meaningful extrapolation.

The disappearance times (DT50) of the test item were:
> 10,000 hrs at pH 4, 7 and 9 resp. at 20°C
174, 565 and 744 hrs at pH 4, 7 and 9 resp. at 50°C
7.67, 8.26 and 13.4 hrs at pH 4, 7 and 9 resp. at 60°C

Description of key information

One study (Reingruber, 2017) was performed according to OECD guideline 111 and regarded as a key study (Klimisch score of 1). Values for disappearance time 50 (DT50) were calculated according to the Gustafson and Holden Model or First-Order Multi-Compartment (FOMC) Model.

The disappearance times (DT50) of the test item were:

> 10,000 hrs at pH 4, 7 and 9 resp. at 20°C

174, 565 and 744 hrs at pH 4, 7 and 9 resp. at 50°C

7.67, 8.26 and 13.4 hrs at pH 4, 7 and 9 resp. at 60°C

Additional information

The determination of the hydrolysis as a function of pH of JNJ-4754724-AAA (T002488) was performed according to the OECD guidelines for the testing of chemicals no. 111: “Hydrolysis as a Function of pH", April 13, 2004. Since for pH 4, 7 and 9 more than 10% hydrolysis was observed after 5 days an additional test was performed.

At pH 4, pH 7 and pH 9 no pseudo-first order reaction kinetic was observed above room temperature. Reaction rates were not constant during the experiment. Accordingly the Arrhenius equation could not be used to determine the rate constant and half-life time at 25°C. Instead, values for disappearance time 50 (DT50) were calculated according to the Gustafson and Holden Model or First-Order Multi-Compartment (FOMC) Model using Computer Aided Kinetic Evaluation (CAKE) Version 3.3. Confidence intervals of the DT50 were back calculated using the 90% Confidence Intervals of the α and β parameter as determined with CAKE. Confidence intervals of the DT50 were not calculated if the DT50 was >10000, as variability and trend of the corresponding data did not allow meaningful extrapolation.

The disappearance times (DT50) of the test item were:

> 10,000 hrs at pH 4, 7 and 9 resp. at 20°C

174, 565 and 744 hrs at pH 4, 7 and 9 resp. at 50°C

7.67, 8.26 and 13.4 hrs at pH 4, 7 and 9 resp. at 60°C