9-Octadecenoic acid (Z)-, monoester with 1,2,3-propanetriol ester with boric acid (H3BO3)

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:

5 November 1982 to 18 November 1982

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Study conducted in compliance with agreed protocols, with no or minor deviations from standard test guidelines and/or minor methodological deficiencies, which do not affect the quality of the relevant results. The study report was conclusive, done to a valid guidelines and the study was conducted under GLP conditions.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1982

Report date:

1982

Materials and methods

Test guideline

Qualifier:

equivalent or similar 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:

9-Octadecenoic acid (Z)-, monoester with 1,2,3-propanetriol ester with boric acid (H3BO3)

EC Number:

264-092-6

EC Name:

9-Octadecenoic acid (Z)-, monoester with 1,2,3-propanetriol ester with boric acid (H3BO3)

Cas Number:

63310-16-7

Molecular formula:

C21H39O5B (based on the representative structure below)

IUPAC Name:

2-hydroxy-3-{[hydroxy({2-hydroxy-3-[(9Z)-octadec-9-enoyloxy]propoxy})boranyl]oxy}propyl (9Z)-octadec-9-enoate; 3-{[bis({2-hydroxy-3-[(9Z)-octadec-9-enoyloxy]propoxy})boranyl]oxy}-2-hydroxypropyl (9Z)-octadec-9-enoate; {2-hydroxy-3-[(9Z)-octadec-9-enoyloxy]propoxy}boronic acid

Test material form:

not specified

Details on test material:

- Physical state: liquid

Method

Target gene:

Histidine synthesis

Species / strain

Species / strain 1

Species / strain / cell type:

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

Additional strain / cell type characteristics:

not applicable

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

0.1, 0.3, 1.0, 3.0 and 10.0 mg/plate both in the presence and absence of metabolic activation

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation). Top agar containing the test material, bacteria, and in some cases S9 mix was mixed and uniformly distributed over the surface of a minimal agar plate. Prior to testing, the molten top agar was prepared by adding 9 x 10^-5 mmoles each of L-histidine and D-biotin. 0.5 mL S9 mix (or S9 buffer) was then added to the number of aliquots of one strain required for one concentration, followed by 0.1 mL of the appropriate concentration of the test substance preparation. Finally 2 mL top agar was then added to each aliquot (2.5 mL for tests without S9 mix), and the resulting mixture poured onto the surface of a prepared pre-labelled plate.

DURATION
- Exposure duration: Plates were incubated inverted at 37 °C for two to three days

NUMBER OF REPLICATIONS: Test concentrations were performed in triplicate

- EVALUATION PROCEDURE: Following the total incubation the number of revertant colonies was counted and the presence and condition of the bacterial lawn was noted.

Evaluation criteria:

Criteria for response in the Ames test are outlined in the field "Any other information on materials and methods incl. tables" section.

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

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

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

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES:
The test material was checked for toxicity on strain TA100 without S9 mix at dose levels of 0.005 to 10 mg/plate. The plates were run in duplicate. No toxicity was observed at these dose levels and so it was decided to test concentrations of test material from 0.1 to 10.0 mg/plate in the definitive study.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Results (Mean values of triplicate plates)

Treatment	Amount/plate	No. revertant colonies per plate
		TA98		TA100		TA1535		TA1537		TA1538
		without S9	with S9	without S9	with S9	without S9	with S9	without S9	with S9	without S9	with S9
Test material	10.0 mg	16	35	85	118	25	15	15	18	14	20
	3.0 mg	25	32	86	102	24	11	16	19	14	23
	1.0 mg	14	36	94	100	26	14	14	18	16	24
	0.3 mg	22	32	113	103	25	13	15	22	21	23
	0.1 mg	22	32	88	85	30	10	18	19	14	23
DMSO	0.1 mL	19	33	90	90	23	14	21	20	13	23
Positive control		2-nitrofluorene (10 µg)	2-aminoanthracene (2 µg)	sodium azide (1 µg)	2-aminoanthracene (2 µg)	sodium azide (1 µg)	2-aminoanthracene (2 µg)	9-aminoacridine (50 µg)	2-aminoanthracene (2 µg)	2-nitrofluorene (10 µg)	2-aminoanthracene (2 µg)
		1964	3765	781	3605	589	254	1300	385	3928	4256

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Under the specific conditions of this assay, the test material gave a negative (i.e. non-mutagenic), response in Salmonella typhimurium strains TA1535, TA1537, TA1538, TA98 and TA100 both in the presence and absence of metabolic activation. No toxicity was observed. The tester strains responded to the positive controls, known mutagens, as expected, confirming the validity of the test method employed. The study is considered to be reliable, relevant and adequate for risk assessment and classification and labelling purposes.

Executive summary:

The potential of the test material to cause gene mutation in bacterial strains was determined following a methodology equivalent to standardised guideline OECD 471. Five strains of Salmonella typhimurium (TA1535, TA1537, TA1538, TA98 and TA100) were treated in the presence and absence of a rat liver derived metabolic activation system (S9 mix). Under the conditions of the study, the test material did not induce any significant, reproducible increases in the observed number of revertant colonies in any of the tester strains used either in the presence or absence of metabolic activation. No toxicity was observed. The tester strains responded to the positive controls, known mutagens, as expected, confirming the validity of the test method employed.