vinasses, residue of fermentation, depotassified

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:

10 Sep - 05 Oct 1992

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

In this study, the substance tested, Vinasses, residue of fermentation, has a chemical composition analogue to Vinasses, residue of fermentation, depotassified and therefore is used in an analogue approach. The analogue approach justification is described in the endpoint study summary. GLP-guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Report date:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Vinasses, residue of fermentation

IUPAC Name:

Vinasses, residue of fermentation

Constituent 2

Reference substance name:

932-215-9

EC Number:

932-215-9

IUPAC Name:

932-215-9

Details on test material:

- Name of test material (as cited in study report): LUCULLUS 235/02
- Physical state / appearance: brown pasty mass
- Analytical purity: 100%
- Lot/batch No.: Lot 235/04
- Storage condition of test material: at +4°C

The test substance is a proteinic basis, whose intended use being for the preparation of reaction flavours.

Method

Target gene:

his operon

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 1537, TA 102, TA 98, TA 100

Metabolic activation:

with and without

Metabolic activation system:

S9 mix, S9 fraction comes from liver homogenates from rats induced with Aroclor 1254 (500 mg/kg)

Test concentrations with justification for top dose:

312.5, 625, 1250, 2500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: distilled water
- Concentration of the test substance in solvent: 50 mg/mL

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: without S9 mix: sodium azide (TA 1535, TA 100), 9-Aminoacridine (TA 1537), 2-Nitrofluorene (TA 98), mitomycin C (TA 102); with S8 mix: 2-Anthramine (TA 1535, TA 1537, TA 98, TA 100), Danthron (TA 102)

Remarks:

with and without S9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: direct in agar (plate incorporation) both tests without S9 mix, first test with S9 mix
preincubation method: second test with S9 mix described by Maron and Ames, 1983

DURATION
- Preincubation period: 60 minutes at 37°C
- Exposure duration: 48 to 72 hours

NUMBER OF REPLICATIONS: 2 per dose

DETERMINATION OF TOXICITY
- Method: relative total growth, preliminary test

Evaluation criteria:

For determining a positive response:
- a reproducible and significant dose relationship using a linear regression analysis, considered as significant if p <= 0.05 (for n = 18 values, the correlation coefficient must be r >= 0.47)
and/or
- a reproducible and significant increase (i.e. a doubling in the number of revertants for at least one of the tested strains when compared to that of the negative and/or solvent controls) for at least one of the tested concentrations.

A test substance is considered as non-mutagenic in this test system if the above two criteria are not fully met.

Statistics:

Biological and statistical significances were considered during the evaluation.

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 102, TA 98, 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

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: preliminary toxicity test (10, 100, 1000, 2500 and 5000 µg/plate)

COMPARISON WITH HISTORICAL CONTROL DATA: yes, in the range

ADDITIONAL INFORMATION ON TOXICITY: The test substance was not toxic at the concentrations used.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

The number of revertants obtained in the presence of LUCULLUS 235/02 - Batch No 235/04, with and without S9 mix, for the 5 strains was equivalent to that of the negative (untreated) and/or solvent controls.

Table 1: Lucullus 235/02 Batch No. 235/04: Results of the first and second mutagenicity test with and without metabolic activation

1st experiment	number of revertants: mean value of negative control	number of revertants: mean value of positive control	max. number of revertants and concentration max. number appeared at [µg/plate]
without S9-mix
TA 1535	9 ± 2	576 ± 20	12 ± 2 [625]
TA 1537	7 ± 1	224 ± 33	10 ± 1 [2500]
TA 102	315 ± 18	1668 ± 88	358 ± 49 [625]
TA 98	18 ± 1	159 ± 7	25 ± 4 [0]
TA 100	115 ± 3	458 ± 13	111 ± 10 [5000]
with S9-mix
TA 1535	12 ± 2	411 ± 57	13 ± 2 [625]
TA 1537	8 ± 2	456 ± 22	10 ± 1 [1250]
TA 102	369 ± 27	931 ± 69	400 ± 12 [5000]
TA 98	25 ± 8	1927 ± 59	21 ± 2 [625]
TA 100	125 ± 2	2104 ± 99	126 ± 12 [1250]
2nd experiment	number of revertants: mean value of negative control	number of revertants: mean value of positive control	max. number of revertants and concentration max. number appeared at [µg/plate]
without S9-mix
TA 1535	9 ± 2	359 ± 17	11 ± 3 [312.5, 625, 5000]
TA 1537	9 ± 2	186 ± 12	13 ± 5 [2500, 5000]
TA 102	226 ± 9	1490 ± 157	295 ± 49 [1250]
TA 98	25 ± 3	170 ± 13	31 ± 4 [0]
TA 100	105 ± 9	603 ± 66	119 ± 18 [0]
with S9-mix
TA 1535	12 ± 2	118 ± 9	14 ± 5 [5000]
TA 1537	7 ± 1	155 ± 19	9 ± 5 [2500]
TA 102	259 ± 3	933 ± 55	395 ± 8 [5000]
TA 98	19 ± 1	969 ± 62	20 ± 3 [5000]
TA 100	94 ± 7	1392 ± 76	111± 17 [1250]

Under the experimental conditions the test substance did not show mutagenic activity in the Ames test.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative