2,2-bis(hydroxymethyl)propane-1,3-diyl didocosanoate

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:

weight of evidence

Study period:

09 Jan - 07 Feb 2002

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP -guideline study with acceptable restrictions (no analytical purity reported)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2002

Report date:

2002

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no analytical purity reported

Test guideline 2

Qualifier:

equivalent or similar to guideline

Guideline:

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

Version / remarks:

adopted in 2000

Deviations:

yes

Remarks:

no analytical purity reported

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C16-18 and C18-unsatd., branched and linear, esters with trimethylolpropane

Cas Number:

403507-18-6

Molecular formula:

not applicable, substance is a UVCB

IUPAC Name:

Fatty acids, C16-18 and C18-unsatd., branched and linear, esters with trimethylolpropane

Details on test material:

- Name of test material (as cited in study report): only trade name given
- Physical state: dark amber, slightly viscous liquid
- Analytical purity: no data
- Stability under test conditions: room temperature, in the dark

Method

Target gene:

his operon

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 102

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with phonobarbitone/beta-naphthoflavone

Test concentrations with justification for top dose:

First and second experiment: 50, 150, 500, 1500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: acetone

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: -S9: N-ethyl-N´-nitro-N-nitrosoguanidine (ENNG, 3 or 5 µg/plate) for TA 100 and TA 1535; 9-Aminoacridine (9AA, 80 µg/plate) for TA 1537; mitomycin C (MMC, 0.5 µg/plate) for TA 102; 4-Nitroquinoline-1-oxide (4NQO, 0.2 µg/plate) for TA98

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: +S9: 2-Aminoanthracene (2AA, 1 or 2 µg/plate) for TA 100, TA 1535 and TA 1537; Benzo(a)pyrene (BP, 5 µg/plate) for TA 98; 1,8-Dihydroxyanthraquinone (DAN, 10 µg/plate) for TA 102

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 replications each in 2 independent experiments

DETERMINATION OF CYTOTOXICITY
- Method: Inspection of the bacterial background lawn

Evaluation criteria:

The test material may be considered positive mutagenic in this test system if the following criteria are met: the test item should induce a reproducible, dose-related and statistically significant increase in the revertant count in at least one strain of bacteria.

Statistics:

Mean values and standard deviation were calculated.

Results and discussion

Test results

Test results 1

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:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1. Test results of experiment 1 (plate incorporation).

 

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	TA102	TA98	TA1537
–	0	80 ± 5.7	24 ± 6.1	294 ± 18.1	24 ± 4.6	18 ± 2.5
–	50	81 ± 6.1	22 ± 5.9	297 ± 19.3	23 ± 1.0	14 ± 2.1
–	150	86 ± 3.1	23 ± 3.2	310 ± 18.2	21 ± 4.6	15 ± 4.7
–	500	82 ± 11.0	23 ± 2.1	330 ± 31.5	22 ± 4.2	19 ± 4.4
–	1500	89 ± 1.5	23 ± 4.0	311 ± 14.0	29 ± 5.7	16 ± 3.2
–	5000	85 ± 5.9 P	30 ± 2.6 P	321 ± 34.6 P	28 ± 4.2 P	16 ± 4.6 P
Positive controls, –S9	Name	ENNG	ENNG	MMC	4NQO	9AA
	Concentrations (μg/plate)	3	5	0.5	0.2	80
	Mean No. of colonies/plate (average of 3 ± SD)	439 ± 33.5	464 ± 29.4	740 ± 113.4	96 ± 5.9	1510 ± 150.1
+	0	92 ± 10.4	25 ± 4.7	339 ± 24.3	40 ± 3.2	23 ± 0.6
+	50	109 ± 11.5	23 ± 7.2	375 ± 14.6	39 ± 9.2	18 ± 2.0
+	150	99 ± 18.0	19 ± 1.5	359 ± 22.4	45 ± 1.2	24 ± 3.1
+	500	94 ± 17.7	15 ± 2.6	358 ± 23.7	45 ± 4.6	26 ± 1.5
+	1500	102 ± 9.6	23 ± 5.6	358 ± 32.1	35 ± 2.0	21 ± 5.2
+	5000	102 ± 6.8	22 ± 4.7	369 ± 72.2	39 ± 4.6	25 ± 2.1
Positive controls, +S9	Name	2AA	2AA	DAN	BP	2AA
	Concentrations (μg/plate)	1	2	10	5	2
	Mean No. of colonies/plate (average of 3 ± SD)	2112 ± 97.7	269 ± 37.8	706 ± 32.6	232 ± 19.2	217 ± 12.4

 

Table 2. Test results of experiment 2 (plate incorporation).

 

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates ± Standard deviation)
		Base-pair substitution type			Frameshift type
		TA 100	TA1535	TA102	TA98	TA1537
–	0	82 ± 8.1	21 ± 4.0	249 ± 20.7	18 ± 0.6	10 ± 2.6
–	50	70 ± 13.1	15 ± 2.6	269 ± 9.8	24 ± 1.5	12 ± 5.5
–	150	79 ± 15.5	18 ± 1.5	279 ± 34.8	24 ± 4.5	6 ± 2.3
–	500	81 ± 7.8	19 ± 6.4	268 ± 2.3	22 ± 6.1	7 ± 1.0
–	1500	88 ± 8.0	22 ± 6.1	275 ± 8.3	27 ± 7.0	13 ± 3.5
–	5000	75 ± 12.1 P	21 ± 3.6 P	290 ± 50.2 P	24 ± 2.9 P	10 ± 6.4 P
Positive controls, –S9	Name	ENNG	ENNG	MMC	4NQO	9AA
	Concentrations (μg/plate)	3	5	0.5	0.2	80
	Mean No. of colonies/plate (average of 3 ± SD)	679 ± 29.4	933 ± 340.6	1167 ± 40.7	188 ± 11.7	2357 ± 179.5
+	0	87 ± 21.1	10 ± 4.4	330 ± 21.2	40 ± 2.1	19 ± 3.8
+	50	86 ± 7.2	13 ± 4.0	369 ± 18.9	33 ± 4.6	17 ± 2.6
+	150	105 ± 10.4	11 ± 4.4	349 ± 27.5	42 ± 1.0	14 ± 7.1
+	500	80 ± 8.0	15 ± 6.1	364 ± 8.3	35 ± 2.5	18 ± 3.6
+	1500	85 ± 6.7	12 ± 3.0	369 ± 34.9	38 ± 2.0	18 ± 1.5
+	5000	88 ± 12.9 P	11 ± 2.5 P	365 ± 15.0 P	36 ± 8.2 P	19 ± 4.4 P
Positive controls, +S9	Name	2AA	2AA	DAN	BP	2AA
	Concentrations (μg/plate)	1	2	10	5	2
	Mean No. of colonies/plate (average of 3 ± SD)	3159 ± 145.2	313 ± 14.5	1021 ± 104.5	312 ± 32.4	222 ± 10.2

 

ENNG = N-ethyl-N-nitro-N-nitrosoguanidine

MMC = mitomycin C

4NQO = 4-nitroquinoline-N-oxide

9AA = 9-aminoacridine

2AA = 2 -aminoanthracene

BP = benzo(a)pyrene

DAN = 1,8 -dihydroxanthraquinone

P = Precipitate

 

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative