Fatty acids, C8-10, oxybis(2-hydroxy-3,1-propanediyl) esters

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 May - 10 June 1999

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1999

Report date:

1999

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

adopted May 26, 1983 (2)

Deviations:

yes

Remarks:

S. typhimurium TA 1538 instead of E. coli or S. typhimurium TA102 tested

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Version / remarks:

dated December 29, 1992

Deviations:

yes

Remarks:

S. typhimurium TA 1538 instead of E. coli or S. typhimurium TA102 tested

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Fatty acids, C8-10, oxybis(2-hydroxy-3,1-propanediyl) esters

EC Number:

295-371-0

EC Name:

Fatty acids, C8-10, oxybis(2-hydroxy-3,1-propanediyl) esters

Cas Number:

92044-91-2

Molecular formula:

UVCB substance - complex ester mixture

IUPAC Name:

Fatty acids, C8-10, oxybis(2-hydroxy-3,1-propanediyl) esters

Test material form:

liquid

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 1538

Metabolic activation:

with and without

Metabolic activation system:

Cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Phenobarbital or Naphthoflavone.

Test concentrations with justification for top dose:

1st experiment : 8; 40; 200; 1000 and 5000 µg/plate
2nd experiment: 20, 60, 200, 600 and 1800 µg/plate

Vehicle / solvent:

The test item was dissolved in DMSO and diluted with DMSO to the desired concentrations just before the start of the test. The solution medium was chosen according to the solubility properties tested preliminary before start of the study.

Controls

Untreated negative controls:

yes

Remarks:

Solution medium DMSO and the untreated fresh cell suspensions served as negative controls.

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

-S9: sodium azide in strains TA 100 and TA 1535, 9-aminoacridine in strain TA 1537, 4-nitro-o-phenylendiamine in strains TA 98 and TA 1538 +S9: 2-aminoanthracene in all strains

Positive control substance:

9-aminoacridine

sodium azide

other: 4-nitro-o-phenylendiamine; 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: triplicate plates in both experiments

DETERMINATION OF CYTOTOXICITY
- Method: mean number of revertant colonies, inspection of bacterial background lawn

Evaluation criteria:

A combination of the following criteria was considered as a positive result:
- The plate background of non-converted 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.

Statistics:

For each tester strain, the mean of the number of revertants and the standard deviations were calculated.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 1538, TA 1537, TA 1535, TA 100 and TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Slightly toxic effects were noted at the concentration of 1000 µg/plate or higher.

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

COMPARISON WITH HISTORICAL CONTROL DATA: the mean revertant number of the vehicle and positive controls was within their respective historical ranges.

Any other information on results incl. tables

Table 1: Experiment 1:

single data of revertant rates without and with metabolic activation

without S9-Mix

strain			TA 100			TA	1535			TA 1537			TA 1538			TA 98
	µg/plate	1	2	3	1		2	3	1	2	3	1	2	3	1	2	3
neg. control		121	115	103	15	8		13	11	13	11	21	12	11	14	23	23
solvent		94	97	90	21	19		14	12	19	8	4	18	10	21	18	22
cone. 1	8	107	101	106	18	12		15	22	12	13	13	16	8	32	31	19
cone. 2	40	77	112	93	8	12		8	13	19	13	12	6	7	19	19	20
cone. 3	200	99	124	89	15	13		12	8	5	15	8	8	6	32	16	30
cone. 4	1000	74	89	110	13	16		8	7	12	8	2	1	4	27	23	19
cone. 5	5000	48	66	32	8	14		11	0	0	0	0	0	0	25	10	13
pos. control	A	887	919	910	585	692		708	858	1407	849	2442	2297	2468	1870	1572	1520
pos. control	B	118	92	158	19	16		15	8	11	15	22	21	16	37	32	21

 

with S9-Mix

strain			TA 100			TA	1535			TA	1537			TA 1538			TA 98
	µg/plate	1	2	3	1		2	3	1		2	3	1	2	3	1	2	3
neg. control		106	119	116	11	8		16	12	18		4	27	18	22	36	30	27
solvent		127	120	83	25	8		14	15	13		13	20	16	22	40	29	29
cone. 1	8	143	108	126	11	15		13	16	13		8	10	16	22	30	33	41
cone. 2	40	120	120	120	17	12		10	15	13		10	14	12	15	35	23	37
cone. 3	200	103	106	105	14	11		13	6	6		8	19	13	15	36	33	33
cone. 4	1000	103	73	108	17	7		10	15	10		14	13	12	8	37	27	35
cone. 5	5000	65	60	69	11	10		16	6	6		1	7	17	11	24	25	35
pos. control	A	232	260	268	207	157		191	472	643		712	2729	2610	2571	2347	2101	2075
pos. control	B	1811	1690	1811	244	274		292	387	321		324	1293	1365	1359	4031	3965	3962

Slightly toxic effects were noted at the concentration of 1000 µg/plate orhigher.

Table 2: Experiment 2:

single data of revertant rates without and with metabolic activation

without S9-Mix

strain			TA 100			TA	1535			TA 1537			TA 1538			TA 98
	µg/plate	1	2	3	1		2	3	1	2	3	1	2	3	1	2	3
neg. control		116	111	120	14	11		5	13	17	8	17	11	13	14	20	24
solvent		108	121	110	13	16		20	15	14	12	12	11	15	24	26	36
cone. 1	20	91	95	102	16	21		16	17	13	19	28	21	20	19	15	25
cone. 2	60	103	107	82	12	14		10	10	8	14	12	10	12	14	19	27
cone. 3	200	126	90	93	19	11		28	15	6	10	6	8	8	27	25	20
cone. 4	600	105	119	81	6	19		18	12	4	16	8	13	6	17	29	14
cone. 5	1800	65	66	67	10	10		15	2	4	5	3	5	3	22	23	22
pos. control	A	858	870	866	671	686		669	1100	1004	1805	1937	1874	1847	1492	1537	1340
pos. control	B	140	130	100	28	15		13	22	12	28	24	31	16	28	27	24

 

with S9-Mix

strain			TA 100			TA	1535			TA	1537			TA 1538			TA 98
	µg/plate	1	2	3	1		2	3	1		2	3	1	2	3	1	2	3
neg. control		122	122	115	8	8		17	10	17		14	12	14	15	19	28	39
solvent		125	98	106	8	21		18	11	22		11	8	23	15	31	37	32
cone. 1	20	125	122	111	16	12		11	10	11		19	27	14	16	35	35	28
cone. 2	60	99	107	113	16	12		20	17	7		15	11	17	18	33	33	37
cone. 3	200	110	121	88	10	12		4	13	22		6	15	14	12	40	32	21
cone. 4	600	97	97	87	17	12		12	7	7		17	5	13	8	30	24	38
cone. 5	1800	82	67	75	14	7		17	10	6		10	7	11	11	25	35	28
pos. control	A	285	330	297	239	276		236	509	855		865	2323	2515	2037	2104	1698	1598
pos. control	B	1534	1368	1644	299	338		326	375	323		344	1435	1501	1527	3782	3720	4105

 

Table 3: Summarizing table of experiment 1 and 2 (mean values and standard deviation (SO))

experiment 1

treatment	strains:
groups      µg/pl.	TA 100 -  (SO) +     (SO)	-	TA 1535 (SO)     +		(SO)	-	TA 1537 (SO)     +		(SO)	-	TA (SO)	1538 +	(SO)	-	TA (SO)	98 +	(SO)
medium	113  9  113     7	12	3	12	4	11	1	11	7	14	6	22	4	20	6	31	5
solvent	94  3  110    24	18	4	16	9	13	5	13	1	11	7	19	3	21	2	33	7
substance 8	105  3  126     18	15	3	13	2	16	6	12	4	12	4	16	6	27	7	35	6
40	94  18 120    0	10	2	13	4	15	4	12	3	8	3	13	2	19	1	32	7
200	104 18 105    2	13	2	12	2	10	5	7	1	8	1	16	3	26	9	34	2
1000	91  18  95    19	12	4	11	5	9	2	13	3	2	2	11	2	23	4	33	6
5000	48  17  65    4	11	3	12	3	0	0	5	3	0	0	12	5	16	8	28	6
positive          A	905 17 253    19	662 17	67 2	185 270	25 24	1038 11	320 3	609 344	123 37	2402 20	92 3	2636 1339	82 40	1654 30	189 8	2174 3986	150 39
controls         B   123 33 1771  70

 

experiment 2

treatment groups            µg/pl.	strains: TA 100 -  (SO)  +     (SO)	-	TA 1535 (SO)     +		(SO)	-	TA 1537 (SO) +		(SO)	-	TA (SO)	1538 +	(SO)	-	TA (SO)	98 +	(SO)
medium solvent	116  4     119     4 113  7     110     14	10 16	4 4	11 16	5 7	13 13	4 2	13 14	4 6	13 12	3 2	13 16	2 7	19 29	5 6	29 33	10 3
substance 20	96  5    119    7	18	3	13	3	16	3	13	5	23	4	19	7	20	5	33	4
60	97  14    106    7	12	2	16	4	11	3	13	5	11	1	15	4	20	7	34	2
200	103 20    106    17	19	8	8	4	10	4	14	8	8	1	13	2	24	3	31	10
600	101 19    93    6	15	7	13	3	11	6	11	6	9	3	9	4	20	8	31	7
1800	66  1    75    7	11	3	13	5	4	2	8	2	4	1	10	2	23	1	29	5
positive           A	865  6     304     23	676 18	9 8	250 321	22 20	1303 21	437 8	743 347	203 26	1886 24	46 7	2292 1487	241 47	1456 26	103 2	1800 3869	268 207
controls         B 123 21 1515 139

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
negative