Reaction mass of 3-(4-methyl-3-pentenyl)cyclohex-3-ene-1-carbonitrile and 4-(4-methyl-3-pentenyl)cyclohex-3-ene-1-carbonitrile

Hydrolysis

Link to relevant study record(s)

Reference

Endpoint:

hydrolysis

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019-05-24 to 2019-05-30

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)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Test Item Name: Azuril
- Chemical Name: Reaction mass of 3-(4-methyl-3-pentenyl)cyclohex-3-ene-1-carbonitrile and 4-(4-methyl-3-pentenyl)cyclohex-3-ene-1-carbonitrile
- Batch/Lot Number: A170421E
- CAS Number: 68084-04-8/21690-43-7
- Appearance: Clear, pale yellow liquid
- Expiry Date: 06 June 2019
- Purity: 99.35%
- Storage Condition: Room temperature (15 – 25°C), protected from humidity (tight closed container) and under inert gas

Radiolabelling:

no

Analytical monitoring:

yes

Details on sampling:

A total of 10 tubes with Test Media (aqueous buffers at pH 4, 7 and 9) were spiked with the Spiking Stock Solution (test item at 10 mg/mL). Five of them were processed immediately to determine the initial concentration of the test item and the other five tubes were incubated at 50°C for 5 days and after the incubation period all the tubes were processed and measured.

Four control tubes were also prepared with Test Media (aqueous buffers at pH 4, 7 and 9) without spiking to check the potential interfering components from the buffer. Two of them were processed immediately to determine the initial concentration of the test item and the other two tubes were incubated at 50°C for 5 days and after the incubation period both control tubes were processed and measured.

Buffers:

Aqueous buffers at pH 4, 7 and 9 were used as Test Media.

Details on test conditions:

Aqueous buffer solutions of different pH values (pH 4, 7 and 9) were treated with the test substance or without (control) and incubated under controlled laboratory conditions. The Test Solutions were kept between 50.1 – 50.5°C. After 5 days, buffer solutions were analysed for the test substance and for hydrolysis.

Duration:

0 h

pH:

4

Temp.:

50 °C

Initial conc. measured:

11.135 mg/L

Duration:

0 h

pH:

7

Temp.:

50 °C

Initial conc. measured:

11.321 mg/L

Duration:

0 h

pH:

9

Temp.:

50 °C

Initial conc. measured:

11.796 mg/L

Number of replicates:

Per each pH, two replicates for controls and five replicates for the test item solution were used.

Positive controls:

no

Negative controls:

yes

Remarks:

Test Media without spiking

Statistical methods:

To determine the test item concentration in the saturated solutions the following calculations were applied:

1. The corrected peak area and the relative peak area were calculated using the following equations:

Corrected Peak Area = Peak Area - Peak Area_Blank

Relative Peak Area = (Corrected Peak Area)/(Peak Area ISTD)

Where:
Corrected Peak Area is the corrected peak area (mVs)
Peak Area is the measured area value of the test item derivative from the individual injection (mVs)
Peak Area_Blank is the average area of the interfering compound from the corresponding Blank chromatograms (mVs)
Relative Peak Area is the relative peak area
Peak Area ISTD is the measured area value of the internal standard from the individual injection (mVs)

2. The average relative peak area from the parallel injections was calculated using the next formula:

Relative Peak Area_Average = Relative Peak Area_1+Relative Peak Area_2)/2

Where:
Relative Peak Area_Average is the average relative peak area of the individual injections
Relative Peak Area_1,2 are the relative peak area values of repeated injections

3. The concentration of the test solution was calculated using the following equation:

c_TestSol = (Relative Peak Area_Average - Intercept))/Slope ∙ AnDil./Enrich ∙ 1000

Where:
c_TestSol is the test item concentration in the test solution (mg/L)
Intercept is the intercept of the calibration curve
Slope is the slope of the calibration curve
AnDil. is the dilution of the analytical sample after the enrichment
Enrich is the enrichment during the sample preparation


The stability was determined:

Stability%= c_0/c_i ∙100%

Where:
c_0 and c_i are the test item concentrations measured on the initial and the end point of the experiment (mg/L)

Transformation products:

no

% Recovery:

101.8

pH:

4

Temp.:

50 °C

Duration:

120 h

% Recovery:

106.4

pH:

7

Temp.:

50 °C

Duration:

120 h

% Recovery:

100.2

pH:

9

Temp.:

50 °C

Duration:

120 h

Remarks on result:

not measured/tested

Details on results:

Reported recovery (%) values correspond to Stability (%) values. The Azuril is hydrolytically stable at 50°C and at pH 4, 7 and 9.

Measured initial concentrations

Sample	cTheroretical(mg/L)	pH	cTestSol(mg/L)	cAverage(mg/L)	% of cTheor	RSD (%)
190-HYD-100	0	4	<LOQ	<LOQ	N/A	N/A
190-HYD-101	0	4	<LOQ
190-HYD-102	10.732	4	11.356	11.135	103.8	1.9
190-HYD-103	10.732	4	11.222
190-HYD-104	10.732	4	11.157
190-HYD-105	10.732	4	10.798
190-HYD-106	10.732	4	11.142
190-HYD-107	0	7	<LOQ	<LOQ	N/A	N/A
190-HYD-108	0	7	<LOQ
190-HYD-109	10.732	7	10.918	11.321	105.5	2.8
190-HYD-110	10.732	7	11.543
190-HYD-111	10.732	7	11.690
190-HYD-112	10.732	7	11.089
190-HYD-113	10.732	7	11.364
190-HYD-114	0	9	<LOQ	<LOQ	N/A	N/A
190-HYD-115	0	9	<LOQ
190-HYD-116	10.732	9	11.223	11.796	109.9	3.3
190-HYD-117	10.732	9	11.882
190-HYD-118	10.732	9	11.848
190-HYD-119	10.732	9	11.717
190-HYD-120	10.732	9	12.313

 Measured concentrations after 5 days

Sample	cTheroretical(mg/L)	pH	cTestSol(mg/L)	cAverage(mg/L)	% of cTheor	RSD (%)
190-HYD-121	0	4	<LOQ	<LOQ	N/A	N/A
190-HYD-122	0	4	<LOQ
190-HYD-123	10.732	4	11.587	11.340	105.7	2.1
190-HYD-124	10.732	4	10.971
190-HYD-125	10.732	4	11.304
190-HYD-126	10.732	4	11.365
190-HYD-127	10.732	4	11.473
190-HYD-128	0	7	<LOQ	<LOQ	N/A	N/A
190-HYD-129	0	7	<LOQ
190-HYD-130	10.732	7	11.681	12.041	112.2	3.1
190-HYD-131	10.732	7	12.408
190-HYD-132	10.732	7	11.780
190-HYD-133	10.732	7	11.858
190-HYD-134	10.732	7	12.479
190-HYD-135	0	9	<LOQ	<LOQ	N/A	N/A
190-HYD-136	0	9	<LOQ
190-HYD-137	10.732	9	11.897	11.815	110.1	1.7
190-HYD-138	10.732	9	11.547
190-HYD-139	10.732	9	11.833
190-HYD-140	10.732	9	12.090
190-HYD-141	10.732	9	11.709

Stability of the test item after 5 days

pH	cTestSol(mg/L)		Stability (%)
	Day-0	Day-5
4	11.135	11.340	101.8
7	11.321	12.041	106.4
9	11.796	11.815	100.2

Validity criteria fulfilled:

yes

Conclusions:

Azuril is hydrolytically stable at 50°C.

Executive summary:

The potential for hydrolysis of Azuril was determined according to an OECD 111 guideline study. Azuril is hydrolytically stable at 50°C and at pH 4, 7 and 9.

Description of key information

The potential for hydrolysis of Azuril was determined according to an OECD 111 guideline study. Azuril is hydrolytically stable at 50°C and at pH 4, 7 and 9.