4-[(4-HYDROXYPYRIMIDIN-2-YL)AMINO]BENZONITRILE

Adsorption / desorption

Link to relevant study record(s)

Reference

Endpoint:

adsorption / desorption: screening

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2016-04-13 to 2016-04-14

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 121 (Estimation of the Adsorption Coefficient (Koc) on Soil and on Sewage Sludge using High Performance Liquid Chromatography (HPLC))

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.19 (Estimation of the Adsorption Coefficient (KOC) on Soil and Sewage Sludge Using High Performance Liquid Chromatography (HPLC))

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: European Commission (EC), Technical Guidance Document on Risk Assessment Part III, Chapter 4.3: “Use of (Quantitative) Structure Activity Relationships ((Q)SARs) - Soil and Sediment Sorption”, 2003.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

See study report

Type of method:

HPLC estimation method

Media:

other: HPLC column

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: M15GC2731
- Expiration date of the lot/batch: 2016-07-23 (retest date)
- Purity test date: 2015-03-29

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: At room temperature protected from light

Radiolabelling:

no

Test temperature:

Column temperature: 35°C ± 1°C

Details on study design: HPLC method:

EQUIPMENT
- Apparatus: Acquity UPLC system (Waters, Milford, MA, USA)
- Type: Acquity UPLC TUV detector (Waters)
- Type, material and diemension of analytical (guard) column: Acquity UPLC HSS Cyano, 100 mm x 2.1 mm i.d., dp = 1.8 µm (Waters)
- UV detection: 210 nm
- Column temperature: 35°C ± 1°C
- Flow: 0.4 mL/min
- Injection volume: 5 µL

MOBILE PHASES
- Mobile phase at pH 4: 55/45 (v/v) methanol/acetate buffer pH 4, 0.1M
- Mobile phase pH 7: 50/50 (v/v) methanol/phosphate buffer pH7, 0.1M
- Solutions at pH 4
* Solution of the unretained compound: A 5.0 g/L stock solution of formamide (99.2%, [75-12-7], Alfa Aesar, Karlsruhe, Germany) in methanol was used. The stock solution was diluted to obtain an end solution of 55/45 (v/v) methanol/0.1 M acetate buffer pH 4.
The formamide blank solution was 55/45 (v/v) methanol/0.1 M acetate buffer pH 4.
* Reference substance solutions: Stock solutions of the reference substances at concentrations of approximately 1 g/L in methanol were used. The stock solutions were diluted to obtain an end solution of 55/45 (v/v) methanol/0.1 M acetate buffer pH 4.
The blank solution for the mixture of reference substances was 55/45 (v/v) methanol/0.1 M acetate buffer pH 4.
* Test solution:
1000 mg/L stock solution of the test item was prepared in DMSO. In order to dissolve the test item the stock solution was ultrasonicated for 10 minutes. The stock solution was diluted to obtain an end solution of 1% DMSO in 55/45 (v/v) methanol/0.1 M acetate buffer pH 4.The final concentration of the test item solution was 10.0 mg/L. The test item blank solution was 1% DMSO in 55/45 (v/v) methanol/0.1 M acetate buffer pH 4.
- Solution at pH7
* Solution of the unretained compound: A 5.0 g/L stock solution of formamide (99.2%, [75-12-7], Alfa Aesar, Karlsruhe, Germany) in methanol was used. The stock solution was diluted to obtain an end solution of 50/50 (v/v) methanol/phosphate buffer pH 7, 0.1 M. The formamide blank solution was 50/50 (v/v) methanol/phosphate buffer pH 7, 0.1 M.
* Reference substance solutions: Stock solutions of the reference substances at concentrations of approximately 1 g/L in methanol were used. The stock solutions were diluted to obtain an end solution of 50/50 (v/v) methanol/0.1 M phosphate buffer pH 7. The blank solution for the mixture of reference substances was 50/50 (v/v) methanol/0.1 M phosphate buffer pH 7.
* Test solution: A 1000 mg/L stock solution of the test item was prepared in DMSO. In order to dissolve the test item the stock solution was ultrasonicated for 10 minutes. The stock solution was diluted to obtain an end solution of 1% DMSO in 50/50 (v/v) methanol/0.1 M phosphate buffer pH 7.The final concentration of the test item solution was 10.0 mg/L. The test item blank solution was 1% DMSO in 50/50 (v/v) methanol/0.1 M phosphate buffer pH 7.

REFERENCE SUBSTANCES
- Identity: Acetanilide; Monuron; 2,5-Dichloroaniline; Naphthalene; Benzoic acid phenylester; Fenthion; Phenanthrene; 4,4’-DDT

REPETITIONS
- Number of determinations: The reference substance and test item solutions were injected in duplicate. Blank solutions were analysed by single injection.

EVALUATION
- Calculation of capacity factors k': from retention time of reference items, the test item, and then onsorbed chemical item
- Determination of the log Koc value: a correlation of log k' versus log Koc of the reference item is plotted using linear regression. The log k' value of the test item is then used to calculate its log Koc value.

Key result

Type:

Koc

Value:

11.6 dimensionless

pH:

4

Remarks on result:

other: Extrapolated value

Key result

Type:

log Koc

Value:

1.06 dimensionless

pH:

4

Remarks on result:

other: Extrapolated value

Key result

Type:

Koc

Value:

5.1 dimensionless

pH:

7

Remarks on result:

other: Extrapolated value

Key result

Type:

log Koc

Value:

0.71 dimensionless

pH:

7

Remarks on result:

other: Extrapolated value

Details on results (HPLC method):

- At pH 4:
* Formamide (t0): tr1 = 0,685min tr2= 0,685min
* Acetanilide: tr1 = 0,876min tr2= 0,944min
* Monuron: tr1 = 1,222min tr2= 1,22min
* 2,5-Dichloroaniline: tr1 = 1,177min tr2= 1,396min
* Naphthalene: tr1 = 1,7min tr2= 1,694min
* Benzoic acid phenylester: tr1 = 1,703min tr2= 2,15min
* Fenthion: tr1 = 2,994min tr2= 2,977min
* Phenanthrene: tr1 = 2,81min tr2= 3,251min
* 4,4’-DDT: tr1 = 9,741min tr2= 9,637min
* The equation of the regression line was: log k’ = 0.359 x log Koc – 0.854 (r = 0.97, n = 16).
* Average retention time for the test substance: 0.916 min

- At pH 7:
* Formamide (t0): tr1 = 0,687min tr2= 0,686min
* Acetanilide: tr1 = 0,955min tr2= 0,947min
* Monuron: tr1 = 1,237min tr2= 1,228min
* 2,5-Dichloroaniline: tr1 = 1,418min tr2= 1,405min
* Naphthalene: tr1 = 1,719min tr2= 1,707min
* Benzoic acid phenylester: tr1 = 2,184min tr2= 2,169min
* Fenthion: tr1 = 3,027min tr2= 3,003min
* Phenanthrene: tr1 = 3,306min tr2= 3,28min
* 4,4’-DDT: tr1 = 9,831min tr2= 9,738min
* The equation of the regression line was: log k’ = 0.347 x log Koc – 0.780 (r = 0.98, n = 16).
* Average retention time for the test substance: 0.887 min

Transformation products:

not measured

The following pKa values in the logarithm range of 1 – 14 for acidic and basic groups in the molecular structure of the test item were calculated using the Perrin calculation method. Based on the calculations, the UPLC analysis was performed at pH 4 and pH 7.

6 -Hydroxy-Pyrimidine (acidic): pKa: 8.61

Validity criteria fulfilled:

yes

Conclusions:

The HPLC method using soil-adsorption-reference data was applied for the determination of the adsorption coefficient (Koc) of JNJ-40370226-AAA (T002487). The extrapolated log Koc values of the test item at pH 4 and pH 7 were determined to be 1.06 and 0.71 (at pH4 and pH7 respectively) using the HPLC estimation method according to OECD guideline 121. The results of the test can be considered reliable without restriction.

Description of key information

This endpoint is covered with a GLP-study (Ciric, 2016) performed according to standard test guideline OECD 121. The log Koc of T002487 was determined to be 0.71 using the HPLC estimation method. The partition coefficients in soil, sediment, suspended matter and raw sewage sludge were calculated from the Koc value and the fraction of organic carbon in the compartment as stipulated in the ECHA guidance Chapter 16 - Table R.16-11.

Key value for chemical safety assessment

Koc at 20 °C:

5.1

Additional information

[LogKoc: 0.71]