Dodecanoic acid, ester with oxybis[propanediol]

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

31 Jan - 10 Feb 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study with acceptable restrictions. Only five S. typhimurium strains and neither the tester strain TA102 nor an additional E. Coli strain were included in the testing.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1995

Report date:

1995

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no E. coli or TA102 was included in the testing.

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted in 1983

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

2,3-dihydroxypropyl laurate

EC Number:

205-526-6

EC Name:

2,3-dihydroxypropyl laurate

Cas Number:

142-18-7

Molecular formula:

C15H30O4

IUPAC Name:

2,3-dihydroxypropyl laurate

Details on test material:

- Name of test material (as cited in study report): only trade name given
- Substance class: surfactant, glyceryl laurate
- Chemical denomination: lauric acid monoglyceride
- Physical state: white solid
- Analytical purity: 100% (active substance)
- Purity test date: 17 Nov 1994
- Lot/batch No.: MD 5094081
- Expiration date of the lot/batch: Mar 1995
- Stability: 12 months (pure)
- Stability in solvent: not given
- Storage condition of test material: at room temperature

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254.

Test concentrations with justification for top dose:

First experiment: 8, 40, 200, 1000 and 5000 µg/plate with and without metabolic activation
Second experiment: 6.25, 12.5, 25, 50 and 100 µg/plate with and without metabolic activation

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Tween 80/bidistilled water (0.1 mL/plate)
- Justification for choice of solvent/vehicle: According to solubility properties which were tested prelininary before start of the study, the suspension medium Tween 80/bidist. water was chosen.

Controls

Untreated negative controls:

yes

Remarks:

untreated fresh cell suspensions

Negative solvent / vehicle controls:

yes

Remarks:

suspension medium Tween 80/bidist. water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: without S9: sodium azide (2 µg/plate) for TA100 and TA1535; 9-aminoacridine (80 µg/plate) for TA1537; 4-nitro-o-phenylendiamine (40 µg/plate) for TA98 and TA1538; with S9: 2 aminoanthracene (2.5 or 5 µg/plate) for all strains

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 replications in 2 independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: clearing or diminution of the background lawn

Evaluation criteria:

A combination of the following criteria was considered as a positive result:
The plate background of non-reverted bacteria did not show any growth reduction versus the respective negative controls.
The spontaneous mutation rates of each tester strain per plate were within the characteristic spontaneous mutation range (see Table 1 under "Any other information on materials and methods incl. tables").
As a rule, the positive control showed mutation rates exceeding the control values of TA100 at least by the factor 2.0 and those of the other tester strains at least by the factor 3.03.
- At more than one dose tested, the test substance caused at least a 2.0-fold increase in comparison with the negative controls in the tester strain TA100. For the other tester strains, an increase in the mutation rate of more than 3.0 above the corresponding negative controls was considered positive.

Reproducibility:
If the test substance induces reverse mutations in only one test, and if this effect cannot be reproduced in one or several repeated tests, the initially positive test data will lose their significance. The criteria for interpretation specified here do not apply absolutely. Other aspects in connection with this in vitro test system may as well be taken into account for the final assessment of the test substance.

Statistics:

Mean values and standard deviation were calculated.

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

from 40 µg/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: precipitations were not noted.

COMPARISON WITH HISTORICAL CONTROL DATA: number of revertants in the negative and vehicle controls (with and without S9) were all within the characteristic spontaneous mutation ranges of the test batches presented in Table 1 under "any other information on materials and methods incl. tables".

ADDITIONAL INFORMATION ON CYTOTOXICITY:
First experiment:
The background lawn was reduced or completely cleared in:
TA100 from 40 µg/plate (-S9) and from 200 µg/plate (+S9)
TA1535 from 40 µg/plate (-S9) and from 200 µg/plate (+S9)
TA1537 from 40 µg/plate (-S9 and +S9)
TA1538 from 40 µg/plate (-S9) and from 200 µg/plate (+S9)
TA98 from 200 µg/plate (+S9 and +S9)

Second experiment:
The background lawn was reduced or completely cleared in:
TA100 from 50 µg/plate (-S9) and from 100 µg/plate (+S9)
TA1535 from 100 µg/plate (-S9 and +S9)
TA1537 from 50 µg/plate (-S9 and +S9)
TA1538 from 50 µg/plate (-S9 and +S9)
TA98 from 100 µg/plate (+S9 and +S9)

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 2. Results of Experiment I.

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		TA 100	TA1535	TA1537	TA1538	TA98
–	Untreated	168	20	13	27	28
–	Vehicle	134	17	14	28	32
–	8	119	16	9	29	28
–	40	87	10	4	T	24
–	200	T	T	T	T	20
–	1000	T	T	T	T	T
–	5000	T	T	T	T	T
Positive controls, –S9	Name	Sodium azide	Sodium azide	9-AA	4-NOPD	4-NOPD
	Concentrations (μg/plate)	2	2	80	40	40
	Mean No. of colonies/plate (average of 3)	474	455	604	1675	1359
+	Untreated	152	22	17	23	51
+	Vehicle	159	18	13	31	38
+	8	126	18	12	23	32
+	40	132	13	12	25	30
+	200	96	15	5	23	27
+	1000	45	T	T	8	10
+	5000	20	T	T	T	T
Positive controls, + S9	Name	2-AA	2-AA	2-AA	2-AA	2-AA
	Concentrations (μg/plate)	5	2.5	2.5	5	5
	Mean No. of colonies/plate (average of 3)	1562	183	87	1339	940

2-AA = 2-aminoanthracene

9-AA = 9-aminoacridine

4-NOPD = 4-nitro-o-phenylendiamine

T = toxic

Table 3. Results of Experiment II.

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		TA 100	TA1535	TA1537	TA1538	TA98
–	Untreated	121	13	12	28	24
–	Vehicle	141	13	9	24	31
–	6.25	135	20	14	26	26
–	12.5	137	19	11	34	26
–	25	112	12	7	30	29
–	50	79	13	8	T	28
–	100	T	T	T	T	20
Positive controls, –S9	Name	Sodium azide	Sodium azide	9-AA	4-NOPD	4-NOPD
	Concentrations (μg/plate)	2	2	80	40	40
	Mean No. of colonies/plate (average of 3)	612	518	457	1741	1449
+	Untreated	128	14	12	26	38
+	Vehicle	129	16	17	27	26
+	6.25	135	14	14	27	42
+	12.5	114	13	12	30	27
+	25	129	17	13	28	26
+	50	119	15	8	21	28
+	100	111	15	12	T	23
Positive controls, + S9	Name	2-AA	2-AA	2-AA	2-AA	2-AA
	Concentrations (μg/plate)	5	2.5	2.5	5	5
	Mean No. of colonies/plate (average of 3)	1591	166	143	1499	780

2-AA = 2-aminoanthracene

9-AA = 9-aminoacridine

4-NOPD = 4-nitro-o-phenylendiamine

T = toxic

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative