2-Propenal, 2-fluoro-3-(4-morpholinyl)-, (Z)-

Hydrolysis

Administrative data

Endpoint:

hydrolysis

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2019-02-05 to 2019-09-05

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Remarks:

GLP compliant study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 111 (Hydrolysis as a Function of pH)

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

(2Z)-2-fluoro-3-(morpholin-4-yl)prop-2-enal

EC Number:

813-789-9

Cas Number:

152873-67-1

Molecular formula:

C7H10FNO2

IUPAC Name:

(2Z)-2-fluoro-3-(morpholin-4-yl)prop-2-enal

Test material form:

solid

Specific details on test material used for the study:

Name: Fluormorpholinacrylaldehyd
Synonym: FMA / (2Z)-2-Fluor-3-(morpholin-4-yl)acrylaldehyd
CAS No.: 152873-67-1
Water Solubility: 260 g/L at 20°C, pH 7 (according to ibacon project No. 127201185)
Batch No.: BXR82PT
Certificate of Analysis/Date: May 29, 2018
Certificate of Analysis/Date (re-analysis): September 04, 2019 (according to ibacon project No. 127201103)
Purity: 86.7%
Purity (re-analysis): 98.6% (according to ibacon project No. 127201103)
Physical Appearance/Colour: Solid, orange-brownish
Retest Date: November 01, 2018
Expiry Date: August 21, 2021 (according to ibacon project No. 127201103)
Storage Conditions at Test Facility: After receipt at the test facility for 8 days at 4°C ± 4 °C until storage recommendations were provided; thereafter at 20°C ± 5°C, in the dark

Radiolabelling:

no

Study design

Analytical monitoring:

yes

Details on sampling:

Preliminary Test (Tier 1)
Test Duration: The incubation was terminated after 5 days.
Sampling of the Test Samples: pH 4, 7 and 9: 0, 3, 24, 120 h
At each sampling point, samples were taken in duplicate.

Main Test (Tier 2)
Test Duration: The incubation at pH 4 was terminated after 25 d (20°C) and 11 d (40°C and 50°C). The incubation at pH 9 was terminated after 30 d (20°C and 40°C) and 22 d (50°C).
Sampling of the Test Samples:
pH 4
20°C: 0 h, 1 d, 2 d, 4 d, 7 d, 11 d, 15 d, 22 d, 25 d
40°C: 0 h, 2 h, 4 h, 6 h, 1 d, 2 d, 3 d, 4 d, 7 d, 11 d
50°C: 0 h, 2 h, 4 h, 6 h, 1 d, 2 d, 3 d, 4 d, 8 d, 11 d
pH 9
20°C: 1 d, 3 d, 7 d, 11 d, 15 d, 22 d, 30 d
40°C: 1 d, 3 d, 7 d, 11 d, 15 d, 22 d, 30 d
50°C: 1 d, 2 d, 3 d, 4 d, 8 d, 11 d, 18 d, 22 d
At each sampling point, samples were taken in duplicate.

Buffers:

Sterile aqueous solutions buffered at pH 4, 7 and 9.
The pH of each buffer solution was measured with a calibrated pH meter.
pH 4: 0.05 M acetate buffer
410 mL acetic acid (0.1 M) was added to 90 mL sodium acetate (0.1 M). The solution was filled up to 1000 mL with pure water.
pH 7: 0.05 M phosphate buffer
296 mL NaOH (0.1 M) was added to 500 mL potassium dihydrogenphosphate (0.1 M). The solution was filled up to 1000 mL with pure water.
pH 9: 0.05 M boric acid buffer
213 mL NaOH (0.1 M) was added to 500 mL of a solution of boric acid (0.1 M) in potassium chloride (0.1 M). The solution was filled up to 1000 mL with pure water.
Anoxic Conditions: To avoid oxidation the buffer solutions were purged with inert gas (nitrogen) prior to sterilisation.
Sterile Conditions: Degased buffer solutions and the used glassware were sterilised using an autoclave (20 min at 121°C) prior to application or heated in a oven (180°C, > 3 h) prior to application.

Details on test conditions:

Sterile aqueous solutions buffered at pH 4, 7 and 9.
Anoxic Conditions: To avoid oxidation the buffer solutions were purged with inert gas (nitrogen) prior to sterilisation.
Sterile Conditions: Degased buffer solutions and the used glassware were sterilised using an autoclave (20 min at 121°C) prior to application or heated in a oven (180°C, > 3 h) prior to application.
Test Units
Test Vessels: Glass flasks (hermetically closed) were used for the test.Test Conditions
Temperature:
Preliminary test (Tier 1): 50°C ± 0.5°C
Main test (Tier 2):
pH 4: 20°C ± 0.6°C; 40°C ± 0.2°C; 50°C ± 0.3°C
pH 9: 20°C ± 0.6°C; 40°C ± 0.6°C; 50°C ± 0.3°C
Light Conditions: The samples were incubated in the dark.
Anoxic Conditions: To avoid oxidation the buffer solutions were purged with inert gas (nitrogen) prior to sterilisation.
Sterile Conditions: Degased buffer solutions and the used glassware were sterilised using an autoclave (20 min at 121°C) prior to application or heated in a oven (180°C, > 3 h) prior to application.
Application
Stock/Application Solutions
Stock Solution of the Test Item: Two individual stock solutions (A and B) were prepared in acetonitrile with a concentration of 2.0 g/L.
Application Solution of the Test Item:
The stock solutions were used for application.
Application of the Test Samples
The final concentration of the test item in the aqueous phase was below 0.01 M or half of its water solubility and the content of organic was 1% v/v. The sample solutions were prepared with a volume of 50 mL and 200 mL and aliquots of 10 mL were transferred to the test vessels for incubation. Each replicate (A and B) was prepared individually.
Application procedure (per replicate):
Tier 1:
pH 4, 7 and 9, Volume stock solution 500 µL, Total sample volume 50 mL, Test item concentration 20 mg/L
Tier 2:
pH 4 and 9, Volume stock solution 2000 µL, Total sample volume 200 mL; Test item concentration 20 mg/L
Preparation of the Blank Samples
Blank Samples:A blank sample for each pH was prepared which consisted of the buffer solution without application of the test item.
Number of Samples: One blank sample of each buffer solution was prepared.
Course of the Study
Aqueous samples were incubated and samples were taken at specific time points.
Preliminary Test (Tier 1)
Test Duration: The incubation was terminated after 5 days.
Main Test (Tier 2)
Test Duration:
The incubation at pH 4 was terminated after 25 d (20°C) and 11 d (40°C and 50°C). The incubation at pH 9 was terminated after 30 d (20°C and 40°C) and 22 d (50°C).
Test Parameters
Temperature: The temperature was recorded continuously.
pH-Value: The pH of the sample solution was determined at each sampling point.
Sterility of the Test Solution: A sterility confirmation test was carried out during Tier 1 and Tier 2. A commercially available dip slide kit was used for a total count of bacteria and to determine the total count of yeast and moulds.

Duration of test

Duration of test 1

Duration:

25 d

pH:

4

Temp.:

20 °C

Initial conc. measured:

20.08 mg/L

Remarks:

Reported initial conc. is mean value of duplicate determination.

Duration of test 2

Duration:

11 d

pH:

4

Temp.:

40 °C

Initial conc. measured:

20.08 mg/L

Remarks:

Reported initial conc. is mean value of duplicate determination.

Duration of test 3

Duration:

11 h

pH:

4

Temp.:

50 °C

Initial conc. measured:

20.38 mg/L

Remarks:

Reported initial conc. is mean value of duplicate determination.

Duration of test 4

Duration:

30 h

pH:

9

Temp.:

20 °C

Initial conc. measured:

20.28 mg/L

Remarks:

Reported initial conc. is mean value of duplicate determination.

Duration of test 5

Duration:

30 d

pH:

9

Temp.:

40 °C

Initial conc. measured:

20.28 mg/L

Remarks:

Reported initial conc. is mean value of duplicate determination.

Duration of test 6

Duration:

22 d

pH:

9

Temp.:

50 °C

Initial conc. measured:

19.7 mg/L

Remarks:

Reported initial conc. is mean value of duplicate determination.

Number of replicates:

Two samples of solutions of each pH value at each test temperature were taken at each sampling point.

Positive controls:

no

Negative controls:

no

Results and discussion

Preliminary study:

Samples were incubated at three different pH-values and at one temperature. The test was carried out for 5 days. During incubation the concentration of the test item remained stable at pH 7 (recovery after 5 days: 95.6% of the nominal applied concentration). In case of pH 4 and 9 recoveries were < LOQ (2 mg/L) and 43.4% of the nominal applied concentration after 5 days, respectively. Thus the test item can be stated as hydrolytically stable at pH 7 and hydrolytically unstable at pH 4 and 9. The main test (Tier 2) was performed at pH 4 and 9.
Values given are averages obtained from duplicate samples.

Test performance:

1) Concerning (Study Plan): 2.2.3 Test Conditions, page 7
According to Study Plan: The temperatures will be maintained to at least ± 0.5°C.
Deviation to Study Plan:
Main test (Tier 2):
pH 4: 20°C ± 0.6°C
pH 9: 20°C ± 0.6°C; 40°C ± 0.6°C
Reason for Deviation: Technical reason during sample placement and incubation.
Presumed Effect on the Study: None. The deviation was of minor extent and only for a short period of time.

2) Concerning (Study Plan): 2.3.2 Application of the Test Samples, page 8
According to Study Plan: Application Procedure:
Main Test (Tier 2):
Per pH level and temperature at least 20 samples.
Deviation to Study Plan: pH 4, 20°C: 18 samples (including 0 h)
pH 9, 20°C: 14 samples
pH 9, 40°C: 14 samples
pH 9, 50°C: 18 samples
Reason for Deviation: Sample number based on the degradation behaviour.
Presumed Effect on the Study: None. Sufficient sampling points were chosen to calculate the DT50 for each pH and temperature.

3) Concerning (Study Plan): 2.7.1 Stock Solutions and Standard Solutions, page 9
According to Study Plan: Standard Solutions: For external standard calibration aliquots of the stock solution will be diluted with pure water/acetonitrile (1/1, v/v).
Deviation to Study Plan: Standard Solutions: For external standard calibration aliquots of the stock solution were diluted with pure water/acetonitrile (1/1, v/v) and buffer (pH 4)/acetonitrile (1/1 v/v) or buffer (pH 9)/acetonitrile (1/1 v/v).
Reason for Deviation: Error.
Presumed Effect on the Study: None. No matrix effect due to the buffer was observed.

4) Concerning (Study Plan): 2.7.3 Analytical Methods, page 10
According to Study Plan: HPLC-Conditions:
Temperature: 30°C
Deviation to Study Plan: HPLC-Conditions:
Temperature: 30°C or room temperature (approx. 22°C)
Reason for Deviation: Error.
Presumed Effect on the Study: None. Peak shape and retention time was equal at room temperature.

Transformation products:

not measured

Total recovery of test substance (in %)

Total recovery of test substance (in %) 1

% Recovery:

32.4

pH:

4

Temp.:

20 °C

Duration:

25 d

Total recovery of test substance (in %) 2

% Recovery:

< 10

pH:

4

Temp.:

40 °C

Duration:

7 d

Remarks on result:

other: % recovery after 7 days was below the limit of quantification

Total recovery of test substance (in %) 3

% Recovery:

< 10

pH:

4

Temp.:

50 °C

Duration:

2 d

Remarks on result:

other: % recovery after 2 days was below the limit of quantification

Total recovery of test substance (in %) 4

% Recovery:

85.5

pH:

9

Temp.:

20 °C

Duration:

30 d

Total recovery of test substance (in %) 5

% Recovery:

< 10

pH:

9

Temp.:

40 °C

Duration:

22 d

Remarks on result:

other: % recovery after 22 days was below the limit of quantification

Total recovery of test substance (in %) 6

% Recovery:

< 10

pH:

9

Temp.:

50 °C

Duration:

11 d

Remarks on result:

other: % recovery after 11 days was below the limit of quantification

Dissipation DT50 of parent compound

Dissipation DT50 of parent compound 1

pH:

4

Temp.:

20 °C

Hydrolysis rate constant:

0.046 d-1

DT50:

15 d

Type:

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

Dissipation DT50 of parent compound 2

pH:

4

Temp.:

40 °C

Hydrolysis rate constant:

0.377 d-1

DT50:

1.84 d

Type:

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

Dissipation DT50 of parent compound 3

pH:

4

Temp.:

50 °C

Hydrolysis rate constant:

1.257 d-1

DT50:

0.552 d

Type:

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

Dissipation DT50 of parent compound 4

pH:

9

Temp.:

20 °C

Hydrolysis rate constant:

0.006 d-1

DT50:

119 d

Type:

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

Dissipation DT50 of parent compound 5

pH:

9

Temp.:

40 °C

Hydrolysis rate constant:

0.063 d-1

DT50:

11.1 d

Type:

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

Dissipation DT50 of parent compound 6

pH:

9

Temp.:

50 °C

Hydrolysis rate constant:

0.188 d-1

DT50:

3.69 d

Type:

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

Details on results:

Samples were incubated at 20, 40 and 50°C. The test was carried out for 25 days (20°C) and 11 days (40/50°C). During incubation the concentration of the test item decreased. At the end of incubation recoveries were 32.4% of the nominal applied concentration at 20°C and < LOQ (2 mg/L) at 40 and 50°C.
Hydrolysis rate constants and corresponding DT50 and DT90 values for the test item were determined for each temperature:
20°C: k = 0.0462 d-1 ;DT50 = 15.0 d; DT90 = 49.9 d
40°C: k = 0.3768 d-1 ;DT50 = 1.8 d; DT90 = 6.1 d
50°C: k = 1.257 d-1 ;DT50 = 0.6 d; DT90 = 1.8 d
The hydrolysis rate constant and the corresponding DT50 value was calculated for a temperature of 25°C based on the arrhenius parameters:
25°C: k = 0.08 d-1 ;DT50 = 8.6 d
Samples were incubated at 20, 40 and 50°C. The test was carried out for 30 d (20°C and 40°C) and 22 d (50°C). During incubation the concentration of the test item decreased. At the end of incubation recoveries were 85.5% of the nominal applied concentration at 20°C and < LOQ (2 mg/L) at 40 and 50°C.
Hydrolysis rate constants and corresponding DT50 and DT90 values for the test item were determined for each temperature:
20°C: k = 0.0058 d-1 ;DT50 = 119.0 d; DT90 = 394.0 d
40°C: k = 0.0625 d-1 ;DT50 = 11.1 d; DT90 = 36.8 d
50°C: k = 0.1878 d-1 ;DT50 = 3.7 d; DT90 = 12.3 d
The hydrolysis rate constant and the corresponding DT50 value was calculated for a temperature of 25°C based on the arrhenius parameters:
25°C: k = 0.011 d-1 ;DT50 = 63.6 d

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

; Preliminary test (Tier 1): 96.8-98.4%; Main test (Tier 2): 98.5-101.9% of the nominal applied concentration

Conclusions:

The test item was found to be hydrolytically stable at pH 7 (DT50 > 1 year at 25°C) and hydrolytically unstable at pH 4 and 9.
The main test (Tier 2) was performed at pH 4 and 9. At the end of incubation at pH 4, recoveries (mean) were 32.4% of the nominal applied concentration at 20°C and < LOQ (2 mg/L) at 40 and 50°C. At pH 9 recoveries were 85.5% of the nominal applied concentration at 20°C and < LOQ (2 mg/L) at 40 and 50°C.

Executive summary:

Title: Fluormorpholinacrylaldehyd: Hydrolysis as a Function of pH [OECD 111]

Test Item: Fluormorpholinacrylaldehyd

Guidelines/Recommendations:

This study was based on the procedures indicated by the following internationally accepted guidelines and recommendations:

OECD Guideline for Testing of Chemicals No. 111: "Hydrolysis as a function of pH", adopted April 13, 2004.

GLP: Yes (certified laboratory)

Purpose: The purpose of the study was to determine the rate of hydrolysis of Fluormorpholinacrylaldehyd at different environmentally relevant pH-values and at different temperatures.

Test Setup: Test Vessels: Glass flasks were used for the test.

Aqueous Solution: Sterile aqueous solutions buffered at pH 4, 7 and 9.

Test Conditions: In the dark at,

Preliminary test (Tier 1): 50°C ± 0.5°C

Main test (Tier 2):

pH 4: 20°C ± 0.6°C; 40°C ± 0.2°C; 50°C ± 0.3°C

pH 9: 20°C ± 0.6°C; 40°C ± 0.6°C; 50°C ± 0.3°C

Treatment Rate: 20 mg/L (two individual replicates)

Results:

The test item was found to be hydrolytically stable at pH 7 (DT50 > 1 year at 25°C) and hydrolytically unstable at pH 4 and 9.

The main test (Tier 2) was performed at pH 4 and 9. At the end of incubation at pH 4, recoveries (mean) were 32.4% of the nominal applied concentration at 20°C and < LOQ (2 mg/L) at 40 and 50°C. At pH 9 recoveries were 85.5% of the nominal applied concentration at 20°C and < LOQ (2 mg/L) at 40 and 50°C. 

 Hydrolysis rate constants and corresponding DT50 and DT90 values for the test item were determined for each temperature:

20°C: k = 0.0462 d-1 ;DT50 = 15.0 d; DT90 = 49.9 d

40°C: k = 0.3768 d-1 ;DT50 = 1.8 d; DT90 = 6.1 d

50°C: k = 1.257 d-1 ;DT50 = 0.6 d; DT90 = 1.8 d

The hydrolysis rate constant and the corresponding DT50 value was calculated for a temperature of 25°C based on the arrhenius parameters: 25°C: k = 0.08 d-1 ;DT50 = 8.6 d

Samples were incubated at 20, 40 and 50°C. The test was carried out for 30 d (20°C and 40°C) and 22 d (50°C). During incubation the concentration of the test item decreased. At the end of incubation recoveries were 85.5% of the nominal applied concentration at 20°C and < LOQ (2 mg/L) at 40 and 50°C.

Hydrolysis rate constants and corresponding DT50 and DT90 values for the test item were determined for each temperature:

20°C: k = 0.0058 d-1 ;DT50 = 119.0 d; DT90 = 394.0 d

40°C: k = 0.0625 d-1 ;DT50 = 11.1 d; DT90 = 36.8 d

50°C: k = 0.1878 d-1 ;DT50 = 3.7 d; DT90 = 12.3 d

The hydrolysis rate constant and the corresponding DT50 value was calculated for a temperature of 25°C based on the arrhenius parameters: 25°C: k = 0.011 d-1 ;DT50 = 63.6 d

This study is classified acceptable and satisfies the guideline requirements for hydrolysis studies.