Pentaerythritol, mixed esters with linear and branched fatty acids

Genetic toxicity: in vitro

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Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

06 May - 29 Aug 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP-Guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

September 1995

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

OECD Guideline 472 (Genetic Toxicology: Escherichia coli, Reverse Mutation Assay)

Version / remarks:

September 1995

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Method

Target gene:

his operon (S. typhimurium strains)
trp operon ( E. coli strains)

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

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

co-factor supplemented post-mitochondrial fraction (S9-mix), prepared from livers of male rats treated with Aroclor 1254

Test concentrations with justification for top dose:

0, 10, 33, 100, 333 and 1000 µg/plate

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

- S9: sodium azide (1 μg/plate, TA1535 and TA100); 9-aminoacridine (75 µg/plate, TA 1537); 2-nitrofluorene (1 µg/plate, TA98 and TA 1538); methylmethanesulfonate (1000 µg/plate, WP2 uvrA); +S9: 2-aminoanthracene (1 μg/pate, all strains)

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 48 to 72 h
- Expression time (cells in growth medium): 48 to 72 h

DETERMINATION OF CYTOTOXICITY
- Method: inspection of the bacterial background lawn wit a dissecting microscope

Evaluation criteria:

Revertant colonies were counted and the mean and standard deviation were calculated and compared to the controls.
All Salmonella tester strains must demonstrate the presence of the deep rough mutation and the deletion of the uvrA gene. Cultures of the TA98 and TA100 strains must demonstrate the presence of the pKM101 plasmid R-factor. All WP2 uvrA cultures must demonstrate the deletion of the uvrA gene. All cultures must demonstrate the characteristic mean number of spontaneous revertants in the vehicle controls. Tester strain titers must be above 30.000.000 cells/ml. The mean of each positive control must be at least three-fold increased to the controlls. A minimum of three non-toxic dose levels are recquired to evaluate assay data.

Statistics:

Mean 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:

other: Unacceptable vehicle control values for the tester strain TA1537 and TA 98 were repeated.

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 1538

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 specified

Positive controls validity:

valid

Test results 3

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity, but tested up to precipitating concentrations

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS

- Precipitation: >100 µg/plate

RANGE-FINDING/SCREENING STUDIES: Yes

COMPARISON WITH HISTORICAL CONTROL DATA: Yes

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:

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		Base-pair substitution type			Frameshift type
		TA 1535	TA1537	TA98	TA100	TA 1538	WP2uvrA
-	Vehicle	12	4	17	111	5	21
-	10	3	4	15	115	6	16
-	33	5	5	19	113	6	11
-	100	9	4	19	98	7	12
-	333	7	5	11	116	4	10
-	1000	8	6	21	120	7	13
Positive controls - S9	Name	SA	9AA	2NF	SA	2NF	MMS
	Concentrations (μg/plate)	1.0	75	1.0	1.0	1.0	1000
	Number of colonies/plate	241	40	105	377	179	143
+	Vehicle	14	5	18	133	7	11
+	10	9	5	27	114	12	16
+	33	9	3	21	113	8	13
+	100	8	7	26	108	9	13
+	333	9	4	27	115	6	8
+	1000	10	5	17	111	9	13
Positive controls + S9	Name	2AA	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	1.0	1.0	1.0	1.0	1.0	10
	Number of colonies/plate	72	127	888	904	783	57

SA: sodium azide

9AA : 9-aminoacridine

MMS: methylmethanesulfonate

2-AA: 2-aminoanthracene

2NF: 2-nitrofluorene

 

 

Table 2: Test results of experiment 2/3:

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 3 plates)
		Base-pair substitution type			Frameshift type
		TA 1535	TA1537	TA98	TA100	TA 1538	WP2uvrA
-	Vehicle	14	10	23	126	11	27
-	10	7	13	16	117	7	27
-	33	9	15	23	124	8	19
-	100	5	13	22	120	6	18
-	333	12	9	17	110	11	16
-	1000	8	14	17	125	5	21
Positive controls - S9	Name	SA	9AA	2NF	SA	2NF	MMS
	Concentrations (μg/plate)	1.0	75	1.0	1.0	1.0	1000
	Number of colonies/plate	429	757	125	601	221	195
+	Vehicle	8	5	19	147	14	24
+	10	9	7	17	142	16	30
+	33	10	5	19	136	18	25
+	100	10	5	20	132	13	31
+	333	10	4	17	138	12	19
+	1000	10	7	19	125	12	19
Positive controls + S9	Name	2AA	2AA	2AA	2AA	2AA	2AA
	Concentrations (μg/plate)	1.0	1.0	1.0	1.0	1.0	10
	Number of colonies/plate	85	97	530	647	1041	88

SA: sodium azide

9AA : 9 -aminoacridine

MMS: methylmethanesulfonate

2-AA: 2 -aminoanthracene

2NF: 2 -nitrofluorene

 

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative