4,4'-Isopropylidenediphenol, oligomeric reaction products with 1-chloro-2,3-epoxypropane, reaction products with [(dimethylamino)methyl]phenol and piperazine

Biodegradation in water: screening tests

Link to relevant study record(s)

Reference

Endpoint:

biodegradation in water: ready biodegradability

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 2016-07-05 to 2016-09-16

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 301 C (Ready Biodegradability: Modified MITI Test (I))

Version / remarks:

1992

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: The Test Method Relating to New Chemical Substances (Japanese notification, Yakushokuhatsu 0331 No. 7, Heisei 23.03.29 Seikyoku No. 5, Kanpokihatsu No. 110331009,

Version / remarks:

21 December 2015

Deviations:

no

GLP compliance:

yes

Specific details on test material used for the study:

Name and physicochemical properties
Name of the new chemical substance (by IUPAC nomenclature) 4,4'-Isopropylidenediphenol, oligomeric reaction products with I-chloro-2,3-epoxypropane, reaction products with [(dimethylamino)methyl]phenol and piperazine
Alternate name THEMIS
CAS RN 159034-96-5
Molecular weight 11,000
Purity (%) of the new chemical substance sub'ected to the study 100.0
Lot number of the new chemical substance subjected to the study 151007
Melting point 120 to 140 ”C
Appearance at room tem erature Powder, pale pink
Stability Swell on exposure to water
Soften greater than or equal to 100”C
Solubility in solvents and the like Solvent Solubility
Dimethylsulfoxide Partially soluble (Visual)
Acetone Almost insoluble (Visual)
Tetrahydrofuran Almost insoluble (Visual)

Oxygen conditions:

anaerobic

Inoculum or test system:

activated sludge (adaptation not specified)

Details on inoculum:

Inoculum: Activated sludge
Source: Chemicals Evaluation and Research Institute, Japan Date of receipt: July 22, 2016 (BOD measurement and recovery test) Mixed liquor suspended solids (MLSS): 4350 mg/L (BOD measurement and recovery test)

Duration of test (contact time):

28 d

Initial conc.:

100 mg/L

Based on:

test mat.

Parameter followed for biodegradation estimation:

O2 consumption

Details on study design:

Test methods
This study was conducted in accordance with the Test Method Relating to New Chemical Substances (Japanese notification, Yakushokuhatsu 0331 No. 7, Heisei 23.03.29 Seikyoku No. 5, Kanpokihatsu No. 110331009, March 31, 2011; the latest revision, December 21, 2015) and OECD Guideline for Testing of Chemicals 301 C "Ready Biodegradability: Modified MITI Test (I)" (1992).
The test substance was exposed to activated sludge in a closed-system oxygen consumption measuring apparatus. The biochemical oxygen demand (BOD) was measured over a 28-day period. After this period, the amount of the dissolved organic carbon (DOC) and the residual amount of the test substance were measured. In addition, GPC, reversed phase HPLC and FT-IR measurement were performed to confirm presence or absence of transformation. The biodegradability of the test substance was evaluated from these results.

Degradability study conditions
This study was conducted under the following conditions.
l) Conditions
Temperature : 25 ± 1”C
Exposure period: 28 days (BOD measurement)
Stirring : Continuous stirring with magnetic stirrer
Test vessel : Culture bottle, glass
C02 absorbent: Soda lime, for carbon dioxide absorption, Kanto Chemical Co., Inc.
Test volume : 300 ml,
Concentration: Test substance, aniline (reference substance): 100 mg/L
Activated sludge: 30 mg/L

2) Test bottle contents and addition of the test substance
Outline of the test solution was shown in the table below. The test substance or reference substance and activated sludge were added to the media in test vessels. The medical paper was used for weighing of the test substance. In addition, to calculate the stirrer bar filing weight during BOD measurement period, the weight of the stirrer bar for using was weighed before BOD measurement.

BOD measurement
The BOD was measured continuously for 28 days.
Apparatus: Closed system oxygen consumption measuring apparatus OM-3100A, Ohkura Electric co. (ID code: Q)

Management and observations during BOD measurement period
In the apparatus, the temperature, stirring, recording meter and air-tightness of the closed system were checked once a day except holidays during BOD measurement period. Observation was made on color of the contents in test bottles using the [Inoculum blank] as a comparative reference. After the end of BOD measurement, observations on color of the contents in test bottles, dissolution state of the test substance and growth of the activated sludge were also made.

Judgment of degradability
The judgment in principle is made in accordance with the following criteria.

1) Readily biodegradable
The biodegradability based on BOD testing is 60% or more in two or more of three test vessels and the average biodegradability of the three test vessels is 60% or more. In addition to the above criterion, it should be confirmed that no degradation products are observed by direct analysis.

2) Non-biodegradable
Properties of not being readily biodegradable

Validity of the test
The test is considered valid if the following criteria are met.
1) The degradability of reference substance based on the BOD exceeds 40% after 7 days and 65% after 14 days, respectively.
2) The difference between the maximum and minimum values of the degradability of the test substance after 28 days is less than 20%.
3) The BOD in the [Inoculum blank] is less than 18 mg 02 (60 mg/L) in 28 days.

Reference substance:

aniline

Remarks:

Chemical name: Aniline Supplier: Kanto Chemical Co., Inc. Grade: Guaranteed reagent Lot number: 709H1215 Purity: 99.8% (Aniline was treated as 100% in purity.)

Key result

Parameter:

% degradation (DOC removal)

Sampling time:

28 d

Remarks on result:

not determinable because of methodological limitations

Key result

Parameter:

% degradation (test mat. analysis)

Value:

>= 0 - <= 7

St. dev.:

0

Sampling time:

28 d

Details on results:

Factors which might have affected the reliability of the test results
There was no specific factor which might have affected the reliability of the test results.

Observation of the contents after the end of BOD measurement
After the end of BOD measurement, observations were made on color of the contents and growth of the activated sludge in the test bottles using the [Inoculum blank] (bottle 6) as a comparative reference, and dissolution state of the test substance.
The content in the [Water + test substance] (bottle 1) and [Sludge + aniline] (bottle 5) were white, and the content in the [Sludge + test substance]-l, 2 and 3 (bottles 2, 3 and 4) were colorless.
Growth of the sludge was observed in the [Sludge + aniline] in contrast with the [Inoculum blank], whereas no growth of the sludge was observed in the [Sludge + test substance].
The test substance was not dissolved visibly in the [Water + test substance] (bottle 1) and the [Sludge + test substance]-l, 2 and 3 (bottles 2, 3 and 4).

pH measurement
At the end of BOD measurement, the pH values were 8.3 for the [Water + test substance] and 7.2, 7.2 and 7.3 for the [Sludge + test substance]-l, 2 and 3, respectively. [Table 1]

Degradability based on the BOD
The BOD *I in the [Water + test substance] after 28 days was 0.0 mg, and the BOD in the [Sludge + test substance]-l, 2 and 3 after 28 days (as corrected with the value in the [Inoculum blank]) were all -0.7 mg. (*1 Theoretical value = 73.7 mg)
The degradabilities based on the BOD after 28 days were calculated to be all 0 % (Calculated values were all -1 %).
The degradabilities based on the BOD after 7 and 14 days were calculated to be 70% and 80%, respectively, for the [Sludge + aniline]. [Table 1]

Degradability based on the DOC
The DOC*2 in the [Water + test substance] after 28 days was 0.9 mg, and the DOC in the [Sludge + test substance]-l, 2 and 3 after 28 days (as corrected with the value in the [Inoculum blank]) were all 0.5 mg. (*2 Theoretical value = 21.6 mg)
The degradability based on the DOC after 28 days was not calculated since the test substance was insoluble in water and the DOC in the [Water + test substance] after 28 days was less than 90% of the theoretical value. [Tables 1 and 2]

Degradability based on the residual test substance amount
The residual amounts of the test substance in the [Water + test substance] after 28 days was 30.6 mg, and that in the [Sludge + test substance]-l, 2 and 3 were 30.2, 30.5 and 28.5 mg, respectively. ( *3 Initial amount = 30.0 mg)
The degradabilities based on the residual amount of the test substance after 28 days were calculated to be 1%, 0%, and 7% for the [Sludge + test substancel-l, 2 and 3, respectively. [Tables 1 and 4]

Confirmation of presence or absence of transformation for NMP soluble component with GPC
The amounts of NMP soluble component in the [Water + test substance] after 28 days was 23.4 mg, and that in the [Sludge + test substance]-l, 2 and 3 were 22.6, 22.3 and 22.7 mg, respectively.
No change was detected in peak shapes in the GPC chromatogram of the test solutions in the [Water + test substance] and the [Sludge + test substance] compared to the peak shape of the standard solution for NMP soluble component. [Tables l and 5]

Confirmation of presence or absence of transformation for NMP soluble component with reversed phase HPLC
The HPLC chromatograms of the [Water + test substance] and the [Sludge + test substance]-l, 2 and 3 were compared to that of the standard solution for NMP soluble component. Although the decrease or increase on the peak tops were observed partially, new peak derived from transformation was not detected on the HPLC chromatogram.

Confirmation of presence or absence of transformation for NMP insoluble component with FT-IR
The obtained results from FT-IR measurement, the infrared radiation spectra of the [Water + test substance] and the [Sludge + test substance]-l, 2 and 3 were compared to that of the standard sample for NMP insoluble component (filtrated residue after vacuum drying). Although interference absorption bands derived from the sludge were observed partially, the characteristic absorption bands with the test substance were detected in the [Water + test substance] and the [Sludge + test substance].

Validity of the test
All of the validity criteria given in the guideline were fulfilled and hence the test was judged valid. [Table 1]

Table 1      Summary of the test results

a) BOD measurement, pH

Bottle No.	Sample description		BOD(mg)				pH
			day 14	day 21	day 28			day 28
	Water + test substance (Abiotic control)	0.0	0.0	0.0	0.0	7.6		8.3
2	Sludge + test substance -l (Test suspension -l)	1.3	1.5	2.7	3.8			7.2
3	Sludge + test substance -2 (Test suspension -2)	1.4	1.5	2.6	3.8			7.2
4	Sludge + test substance -3 (Test suspension -3)	1.2	1.3	2.5	3.8			7.3
5	Sludge + aniline (Activity control)		58.7	62.8	69.1			7.7
6	Inoculum blank (Control blank)		1.3	2.9	4.5	7.0		7.2

b) Measured values (day 28)

		Sludge + test substance			Water + test substance	Theoretical value
		Bottle 2	Bottle 3	Bottle 4	Bottle I
BOD'I	mg	-0.7	-0.7	-0.7	0.0	73.7
DOC"	mg	0.5	0.5	0.5	0.9	21.6
Test substance (Total dry weight)	mg	30.2	30.5
		101	102	95	102
NMP soluble component (GPC)	mg	22.6	22.3	22.7

*l : Value of [Sludge + test substance] is corrected with BOD or DOC value of [Inoculum blank].

c) Degradability (%)

		Sludge + test substance			Average
		Bottle 2	Bottle 3	Bottle 4
BOD'2
DOC"
Test substance				7	3

*2 : Where % degradability is calculated to be negative, this value is shown in parentheses.

*3 : Degradability is not calculated because the DOC in [Water + test substance] is less than 90% of theoretical value.

Table 2 Result of DOC Measurement

Bottle No.		Sample description	Added amount mg	Theoretical value mgC	DOC in bottle mgC	DOCs DOCb mec	Ratio to theoretical value	Degradability
		Water + test substance (Abiotic control)	30.0	21.6	0.9		4	*NA
2		Sludge + test substance-I (Test suspension -l )	30.0	21.6		0.5	2	*NA
3		Sludge + test substance-2 (Test suspension -2)	30.0	21.6		0.5	2	*NA
4		Sludge + test substance-3 (Test suspension -3)	30.0	21.6		0.5	2	*NA
5		Sludge + aniline (Activity control)
6		Inoculum blank (Control blank)			0.6
		Average (Bottle 2,3,4)					2

Equations:                                                                                                        * Not available

Theoretical value : Added amount(mg) x Carbon ratio

             Carbon ratio in the test substance            

Ratio to theoretical value :

[Sludge + test substance] —-100

         [Water + test substance] = (DOCc)+Theoretical valuex 100        

Table 3 Residual test substance amount in test bottles

Bottle No.	Sample description		c
		Total dry mg	Membrane filter Stirrer weight mg	bar filing Test "Eight mg	substance in bottle mg	Added amount mg	Residual test substance rate	Degradability
	Water + test substance (Abiotic control)		63.6	34. I	30.6	30.0	102
2	Sludge + test substance-I (Test suspension -l )		60.5	10.0	30.2	30.0	101
3	Sludge + test substance-2 (Test suspension -2)		60.5	2.8	30.5	30.0	102
4	Sludge + test substance-3 (Test suspension -3)		59.4	18.8	28.5	30.0	95	7
5	Sludge + aniline (Activity control)
6	Inoculum blank (Control blank)	79.5	63.3	6.9	9.3
	Average (Bottle 2,3,4)				29.7		99	3

Equations:

D=A-B-C (Bottle I and 6)

D=A-B.C-D of Bottle 6 (Bottle 2 to 4)

Degradability(%)=(l-Cs/Cc) x 100

Where Cs: amount (mg) in [Sludge + test substance]

Cc: amount (mg) in [Water + test substance]

Table 5 NMP soluble component amount in test bottles

Bottle No.		Sample description		Peak area mV • sec		NMP soluble component in bottle mg
		Water + test substance (Abiotic control)		3993.328		23.4
2		Sludge + test substance-I (Test suspension -l)		3866.852		22.6
3		Sludge + test substance-2 (Test suspension -2)		3821.843		22.3
4		Sludge + test substance-3 (Test suspension -3)		3875.533		22.7
5		Sludge + aniline (Activity control)
6		Inoculum blank (Control blank)				<0.2
		Detection limit		21		0.2

 

	Standard solution		Concentration E :				2320 mg/L
			Peak area		F:		4052.676 mV•sec
	Volume of solution for analysis				G :		0.010 L
	Volume of test solution for analysis				H :		0.300 L
	Volume of test solution in bottle				1 :		0.300 L
	Average recovery						97.9 0/0

Equations:

B=A+FxExGx (I*H) * (J*100)

Detection limit=F>

Detection limit (mg) was derived by rounding B up to first decimal place.

Validity criteria fulfilled:

yes

Interpretation of results:

not readily biodegradable

Conclusions:

It is concluded that the test substance is not readily biodegradable and no transformation product is generated under the conditions of this test

Executive summary:

This study was conducted in accordance with the OECD 301C to assess the Ready Biodegradability of test item.  The test substance was exposed to activated sludge in a closed-system oxygen consumption measuring apparatus. The biochemical oxygen demand (BOD) was measured over a 28-day period. After this period,theamount of the dissolved organic carbon (DOC) and the residual amount of the test substance were measured. In addition, GPC, reversed phase HPLC and FT-IR measurement wereperformed toconfirmpresence or absence of transformation. The biodegradability of the test substance was evaluated from these results.

The test substance is not readily biodegradable and no transformation product is generated under the conditions of this test.

Description of key information

The ready biodegradability was assessed in accordance with OECD 301C.

It is concluded that the test substance is not readily biodegradable and no transformation product is generated under the conditions of this test.

Key value for chemical safety assessment

Biodegradation in water:

under test conditions no biodegradation observed

Type of water:

freshwater