Butyl glycollate

Genetic toxicity: in vitro

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:

1996-MAR-06 through 1996-MAR-28

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Cross-reference

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: U.S. EPA: 798,5265, The Salmonella typhimurium reverse mutation assay Fed. Reg. 50, Subpart F, September 1985

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Butyl glycollate

EC Number:

230-991-7

EC Name:

Butyl glycollate

Cas Number:

7397-62-8

Molecular formula:

C6H12O3

IUPAC Name:

butyl glycolate

Constituent 2

Reference substance name:

Polysolvan O

IUPAC Name:

Polysolvan O

Constituent 3

Reference substance name:

Glycolic acid-n-butyl ester

IUPAC Name:

Glycolic acid-n-butyl ester

Details on test material:

- Name of test material (as cited in study report): Polysolvan O (butyl glycollate)
- Physical state: liquid
- Analytical purity: 98.1%
- Impurities (identity and concentrations): not quantified
- Purity test date: 22-February-1996
- Lot/batch No.: Gef. 34
- Expiration date of the lot/batch: 20-August-1996
- Stability under test conditions: stable for 4 h according to analytical certificate
- Storage condition of test material: darkness at about 5 °C in a refrigerator

Method

Target gene:

Different loci of different bacterial histidine operons.

Species / strain

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

- Type and identity of media:
- Properly maintained: yes
- Periodically checked for karyotype stability: yes

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Supernatant of rat liver homogenate (S9) from Aroclor 1254 induced male Sprague-Dawley rats.

Test concentrations with justification for top dose:

Both experiments:
+S9 mix: 0, 4, 20, 100,, 500, 2500, 5000 µg/plate
-S9 mix: 0, 4, 20, 100,, 500, 2500, 5000 µg/plate

Vehicle / solvent:

- Vehicle: DMSO
- Justification for choice of solvent/vehicle: soluble and stable in DMSO

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

Positive controls:

yes

Remarks:

Sodium azide, 9-Aminoacridine, 2-Nitrofluorene, 2-Aminoanthracene

Positive control substance:

other:

Remarks:

Positive control plates were included for each strain

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h at 37 °C
- Expression time (cells in growth medium): 48 h
- Selection time (if incubation with a selection agent): 48 h
- Fixation time (start of exposure up to fixation or harvest of cells): 48 h

SELECTION AGENT: Histidine

NUMBER OF REPLICATIONS: 3

NUMBER OF EXPERIMENTS: 2

DETERMINATION OF CYTOTOXICITY
- Method: reduced rate of spontaneous occurring colonies

Evaluation criteria:

According to OECD 471 and EU method

Statistics:

None

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:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

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

Results: Experiment 1: Number of mean revertant colonies per plate in S. typhimurium strains

 

Dose µg/plate	TA100	TA1535	1537	98
	With metabolic activation
0	141.3	12.7	8.7	26.0
4	120.3	11.0	7.3	20.7
20	133.7	9.7	9.3	27.0
100	137.0	9.7	8.7	22.0
500	135.3	11.7	9.3	24.3
2500	121.3	10.3	8.0	29.3
5000	142.7	10.0	9.0	29.7
	Without metabolic activation
0	126.0	11.3	7.3	21.7
4	120.3	10.0	9.0	23.7
20	133.7	10.0	7.0	22.0
100	137.0	9.0	8.3	26.7
500	135.3	10.0	8.3	22.7
2500	121.3	8.0	8.3	23.0
10000	142.7	11.0	7.3	19.0

 

Results: Experiment 2: Number of mean revertant colonies per plate in S. typhimurium strains

 

Dose µg/plate	TA100	TA1535	1537	98
	With metabolic activation
0	122.3	10.7	10.0	28.7
4	136.0	8.0	8.7	27.3
20	124.0	11.3	8.7	32.7
100	127.3	10.3	8.3	22.7
500	135.3	8.3	8.0	26.0
2500	138.0	9.7	8.0	30.3
5000	139.0	11.7	9.7	29.3
	Without metabolic activation
0	124.3	11.0	9.7	22.3
4	132.3	11.7	8.7	26.0
20	141.7	12.3	8.0	27.0
100	124.3	10.7	9.7	26.3
500	141.3	12.3	10.0	28.7
2500	129.7	15.0	9.0	22.7
10000	134.0	12.7	7.7	24.7

 

Sterility of S9-mix and the test compound were indicated by the absence of contamination of the test material and S9-mix sterility check plates. Control plates (background control and positive controls) gave the expected number of colonies.

 

The test compound did not precipitate on the plates up to the highest investigated dose of 5000 µg/plate.

Applicant's summary and conclusion

Conclusions:

In the presence or absence of a metabolic activation, Polysolvan 0 did not led to any relevant increase in the number of revertant colonies in the bacterial tester strains.

Executive summary:

Polysolvan O (butyl glycollate, 98.1%) was investigated for its potential mutagenic activity in the in vitro reverse gene mutation assay in bacteria with the strains TA100, TA1535, TA1537 and TA98 of Salmonella typhimurium. The test was performed in the absence and presence of a metabolizing system derived from rat liver homogenate (male Sprague-Dawley rat liver S9-mix induced by Aroclor 1245).

Polysolvan O (batch Gef. 34, liquid, purity: 98.1%) was dissolved in dimethyl sulfoxide (DMSO) and 2 independent experiments with 3 replicate each were performed. The solvent served also as negative control and appropriate positive controls were used. The doses for both studies ranged from 4 to 5000 µg/plate

 

This study is assessed as appropriate and valid since it was performed according to internationally accepted OECD and EU testing guidelines and according to GLP. Reporting, assessment and data presentation in the study report was considered as appropriate.

 

Control plates without mutagen showed that the number of spontaneous revertant colonies was similar to that described in the literature. All the positive control compounds led to the expected increase in the number of revertant colonies.

 

Polysolvan O (butyl glycollate, 98.1%) was not toxic to the bacterial strains up to the highest concentration of each 5000 µg/plate in the presence or absence of metabolic activation.

 

No bacteriotoxicity was found in the toxicity test using histidine enriched agar plates and a dilution of the tester strain TA100 (designated TA100 D), which was performed in parallel with the second experiment, either in the absence or in the presence of metabolic activation.

 

In the absence or presence of a metabolic activation, Polysolvan 0 did not led to any relevant increase in the number of revertant colonies in the bacterial tester strains.

 

Conclusion:

Polysolvan O (butyl glycollate, 98.1%) was not mutagenic in the in vitro reverse gene mutation assay in bacteria with the strains TA100, TA1535, TA1537 and TA98 of Salmonella typhimuriuma in the absence and presence of metabolic activation.