Glycine, N,N'-1,2-ethanediylbis-, reaction products with formaldehyde, iron chloride (FeCl3) and phenol, potassium salts

Hydrolysis

Link to relevant study record(s)

Reference

Endpoint:

hydrolysis

Type of information:

experimental study

Adequacy of study:

key study

Study period:

from 2020-03-05 to 2020-03-10

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:

no

Qualifier:

according to guideline

Guideline:

other: Council Regulation (EC) No. 440/2008, Method C.7 (2008)

Deviations:

no

Principles of method if other than guideline:

None

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Batch number FC-C 11140

CAS number 1463474-95-4

Purity (certified) 96.6% (UVCB)

Appearance Dark red-brown microgranules (solid)

Water solubility 50 – 100 g/L

pH 7 – 9 (1% in water)

Stability under test Not specified
conditions

Expiry date 2022-08-31

Recommended storage Keep in a dry place. Store at room temperature in the original container. Keep container tightly closed.

Receipt 2019-09-12

Identification parameters Name, batch number, state and color

Retention sample At least 1 g has been retained on 2019-09-16 and stored at 6 ± 2 °C.

Storage condition 18 – 25 °C, dark, in tightly closed original container

Radiolabelling:

no

Analytical monitoring:

yes

Details on sampling:

Type and Frequency of Measurements
Samples were taken at test start (0 h) and test end (120 h). All samples were analysed immediately (if possible, less than 30 min after sampling until start of analyses, but at least not more than 2.5% of the total study time) via HPLC-DAD as specified in Annex I of the report. The method was validated according to Annex II of the report.
The incubation temperature was checked automatically once per hour and at least once per day manually.

Preparation of the test samples After sampling, the samples were analysed directly.

Buffers:

Buffer solution pH 4 0.18 g of sodium hydroxide and 5.7555 g of mono potassium citrate were dissolved in 500 mL purified water.

Buffer solution pH 7 3.854 g of ammonium acetate were dissolved in 500 mL purified water.

Buffer solution pH 9 0.426 g sodium hydroxide, 1.8638 g potassium chloride and 1.5458 g boric acid were dissolved in 500 mL purified water.

Details on test conditions:

Replicates Duplicates per pH and sampling date, single injections

Test container (sterile) HPLC vials, brown glass, volume: 2 mL

Test volume 1 mL

Test concentration / solution 150 mg/L (pH 4, 7 and 9)

Application
The test solutions were prepared at test start by direct weighing.
The test item was dissolved in the respective buffer and sterilized by filtration through 0.20 µm sterile membrane filters. After sterilization of the test solutions only sterilized glassware and additional material was used.

After the vials were sealed they were transferred into the thermostat. The time between test item application and transfer to thermostat did not exceed 30 min.

Incubation time 120 h

Temperature 50 ± 0.5 °C

Light Photolytic effects were avoided by using an opaque water bath.
Samples were kept in the dark.

Duration:

120 h

pH:

4

Temp.:

50 °C

Initial conc. measured:

136 mg/L

Duration:

120 h

pH:

7

Temp.:

50 °C

Initial conc. measured:

149 mg/L

Duration:

120 h

pH:

9

Temp.:

50 °C

Initial conc. measured:

145 mg/L

Number of replicates:

Duplicates per pH and sampling date, single injections

Positive controls:

no

Negative controls:

no

Statistical methods:

Not applicable substance is hydrolytically stable.

Preliminary study:

Substance is hydrolytically stable

Hydrolysis Results

Summary of Hydrolysis Results for HBED-Fe at 50 °C

Mean concentration of duplicates at pH 4.0: at T0 = 145 mg/L and at T120h = 149 mg/L
Mean concentration of duplicates at pH 7.0: at T0 = 149 mg/L and at T120h = 151 mg/L
Mean concentration of duplicates at pH 9.0: at T0 = 145 mg/L and at T120h = 149 mg/L

[mg/L] Loss of
Test Item
[%]
0 1 136 136 -
2 135
120 1 138 139 0
2 139

Transformation products:

no

Details on hydrolysis and appearance of transformation product(s):

Substance is hydrolytically stable. No transformation products are formed.

% Recovery:

> 100

pH:

4

Temp.:

50 °C

Duration:

120 h

Remarks on result:

hydrolytically stable based on preliminary test

% Recovery:

> 100

pH:

7

Temp.:

50 °C

Duration:

120 h

Remarks on result:

hydrolytically stable based on preliminary test

% Recovery:

> 100

pH:

9

Temp.:

50 °C

Duration:

120 h

Remarks on result:

hydrolytically stable based on preliminary test

Key result

pH:

4

Temp.:

50 °C

Remarks on result:

hydrolytically stable based on preliminary test

Key result

pH:

7

Temp.:

50 °C

Remarks on result:

hydrolytically stable based on preliminary test

Key result

pH:

9

Temp.:

50 °C

Remarks on result:

hydrolytically stable based on preliminary test

Other kinetic parameters:

not applicable substance is hydrolyticallly stable.

Details on results:

The validity criteria were fulfilled:
The test temperature was 50 ± 0.5°C.
The pH of the buffer solutions were in the range of ± 0.1 pH at test temperature.
The sensitivity of the analytical method was sufficient to analyse test item concentrations at least down to 90 % reduction of the initial concentration.

Loss of Test Item [% of applied] of HBED-Fe at 50 °C after 120 Hours

Hydrolysis Time
[Hours] Loss of Test Item [%]
pH 4 pH 7 pH 9
120 0 0 0

Results with reference substance:

-

Hydrolysis Results for HBED-Fe at pH 4 and 50 °C

Hydrolysis Time [Hours]	Replicate	Concentration [mg/L]	Mean [mg/L]	Loss of test item %
0	1	136	136	-
0	2	135
120	1	138	139	0
120	2	139

Hydrolysis Results for HBED-Fe at pH 7 and 50 °C

Hydrolysis Time [Hours]	Replicate	Concentration [mg/L]	Mean [mg/L]	Loss of test item %
0	1	149	149	-
0	2	149
120	1	151	151	0
120	2	151

Hydrolysis Results for HBED-Fe at pH 9 and 50 °C

Hydrolysis Time [Hours]	Replicate	Concentration [mg/L]	Mean [mg/L]	Loss of test item %
0	1	145	145	-
0	2	145
120	1	149	149	0
120	2	149

Validity criteria fulfilled:

yes

Conclusions:

The test item HBED-Fe was concluded to be hydrolytically stable under all of the tested conditions .

Executive summary:

Hydrolysis as a function of pH was determined according to OECD Guideline No. 111 and Council Regulation (EC) No. 440/2008, Method C.7 for the test item HBED-Fe (batch number: FC-C 11140) from 2020-03-05 to 2020-03-10 at Noack Laboratorien, 31157 Sarstedt, Germany.

Analyses of the test item HBED-Fe were performed via HPLC DAD on a reversed phase analytical column using the test item as external standard. The analytical method was validated with satisfactory results with regard to linearity, accuracy, precision and specificity.

The study was conducted with a test item concentration of 150 mg/L in buffer solutions at pH 4, 7 and 9 at a test temperature of 50 °C (preliminary test). Samples were taken at test start (0 hours) and test end (120 hours). Pure test systems (buffer solution pH 4, 7 and 9) were analysed at test start and test end and there was no analytical interference with the test item.

No significant transformation was observed and therefore, the test item was considered as hydrolytically stable under this condition and a half-life of > 1 year could be assumed for environmental typical temperatures.

Loss of Test Item [% of applied] of HBED-Fe at 50 °C after 120 Hours

Hydrolysis Time [Hours]	pH 4	pH 7	pH
120	0	0	0

Description of key information

HBED is observed to be hydrolytically stable in a preliminary test performed according to OECD 111. Identical results were observed in the supporting study with the analogue substance EDDHMA-FeK.

Key value for chemical safety assessment

Half-life for hydrolysis:

365 d

at the temperature of:

12 °C

Additional information

No hydrolysis was observed for HBED-Fe. The half-life for hydrolysis at 12 deg C for the three pH's tested (4, 7 and 9) is > one year.