Dodecylphosphonic acid

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

between 18 March 1997 and 7 April 1997

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

Guideline study to GLP but TA102 or E.Coli were not tested.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Report date:

1997

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

: TA 102 or E.coli were not tested

Test guideline 2

Qualifier:

according to guideline

Guideline:

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

Deviations:

not specified

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:

Octylphosphonic acid

EC Number:

225-218-5

EC Name:

Octylphosphonic acid

Cas Number:

4724-48-5

Molecular formula:

C8H19O3P

IUPAC Name:

octylphosphonic acid

Test material form:

other: solid

Details on test material:

- Name of test material (as cited in study report): Octylphosphonic Acid (with no residual ester):

Method

Target gene:

Histidine for Salmonella

Species / strain

Species / strain / cell type:

other: S. typhimurium strains TA1535, TA1537, TA1538, TA98, TA100

Details on mammalian cell type (if applicable):

Not applicable

Metabolic activation:

with and without

Metabolic activation system:

Rat S9 mix

Test concentrations with justification for top dose:

5 to 5000 µg/plate

Vehicle / solvent:

acetone

Controls

Controls 1

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

acetone

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene (2AA)

Remarks:

for all strains with metabolic activation

Controls 2

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

acetone

True negative controls:

no

Positive controls:

yes

Positive control substance:

N-ethyl-N-nitro-N-nitrosoguanidine

Remarks:

for TA100 and TA1535 without metabolic activation

Migrated to IUCLID6: ENNG

Controls 3

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

acetone

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

for TA1537 without metabolic activation

Migrated to IUCLID6: 9AA

Controls 4

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

acetone

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-Nitro-o-phenylenediamine (4NOPD)

Remarks:

for TA1538 without metabolic activation

Controls 5

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

acetone

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

Remarks:

for TA98 without metabolic activation

Migrated to IUCLID6: 4NQO

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: no
- Exposure duration: 48h at 37°C

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: the plates were assessed for revertant colonies using a Domino colony counter and examined for a thinning of the background lawn.

Evaluation criteria:

For a substance to be considered positive in this test system, it should have induced a dose-related and statistically significant increase in mutation rate in one or more strains of bacteria in the presence and/or absence of the S9 microsomal enzymes in both experiments at sub-toxic dose levels. If the experiments give conflicting results, a third experiment may be used to confirm the correct result.
To be considered negative, the number of induced revertants compared to spontaneous retertants should be less than twofold at each dose level, the intervals of which should be between 2 and 5 fold and extend to the limits imposed by toxicity, solubility or up to the maximum recommended dose of 5000 µg/plate.

Statistics:

All data are statistically analysed using the methods recommended by the UKEMS and normally Dunnett’s method of linear regression is used to evaluate the result.

Results and discussion

Test results

Species / strain:

other: S. typhimurium TA1535, TA1537, TA1538, TA98, TA100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

from 1500 ug/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS: no data


RANGE-FINDING/SCREENING STUDIES:
Due to the acidic nature of the test material, the preliminary toxicity assay was performed with TA100 both with and without S9-mix and with a dose range of 5 to 5000 µg/plate. The test material exhibited toxicity at and above 1500 µg/plate.

COMPARISON WITH HISTORICAL CONTROL DATA: no data

ADDITIONAL INFORMATION ON CYTOTOXICITY:
The test material caused a visible reduction in the growth of the bacterial background lawn to all of the tester strains, both with and without metabolic activation. The first indication of a toxic response was observed at 1500 µg/plate.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Experiment 1

 

Without activation

Test Group	Dose Level (µg/plate)	Revertant Colony Counts
		TA 1535		TA 1537		TA 1538		TA 98		TA100
		mean	SD	mean	SD	mean	SD	mean	SD	mean	SD
DMSO	Vehicle control	24	3.6	10	2.9	32	3.5	28	7.1	116	5.5
Test	5	27	5.5	9	3.8	36	1.0	27	1.7	112	1.5
	15	30	1.0	10	1.7	34	3.1	23	3.8	113	6.8
	50	29	2.1	7	3.5	36	2.5	19	5.8	117	13.6
	150	29	3.6	6	1.2	29	6.5	24	2.0	123	4.6
	500	27	4.4	5	1.0	27	1.0	20	6.2	103	9.9
	1500	24	5.5	4	2.5	23	4.9	20	3.6	0	0.0
	5000	15	4.0	3	0.6	0	0.0	13	2.1	0	0.0
ENNG	5.0	208	18.9
	3.0									476	47.0
9AA	80			742	19.5
4NOPD	5.0					680	90.1
4NQO	0.2							376	66.8

 

With activation

Test Group	Dose Level (µg/plate)	Revertant Colony Counts
		TA 1535		TA 1537			TA 1538		TA 98		TA100
		mean	SD	mean	SD		mean	SD	Mean	SD	mean	SD
DMSO	Vehicle control	20	4.6	12	1.0		36	3.5	32	2.1	115	2.6
Test	5	19	2.6	15	2.3		33	0.0	34	4.5	105	11.9
	15	16	0.6	11	2.3		28	1.5	27	5.7	130	8.1
	50	22	2.0	11	0.0	32		1.5	29	6.0	120	14.8
	150	23	5.0	13	1.5	32		2.3	26	3.2	123	3.2
	500	19	3.1	11	1.5	33		6.0	31	3.8	115	8.4
	1500	9	2.1	11	1.2		34	5.7	19	4.9	14	2.5
	5000	8	4.2	6	0.6		2	1.2	5	1.0	0	0.0
2AA	2.0	244	17.7	257	71.7
	1.0										845	74.3
	0.5						237	5.5	304	37.3

 

Experiment 2

 

Without activation

Test Group	Dose Level (µg/plate)	Revertant Colony Counts
		TA 1535		TA 1537		TA 1538		TA 98		TA100
		mean	SD	mean	SD	mean	SD	mean	SD	mean	SD
DMSO	Vehicle control	23	4.7	9	2.6	30	6.0	22	5.3	116	4.9
Test	15					34	9.9			114	2.6
	50	22	9.2	9	0.0	33	5.5	20	2.0	116	6.9
	150	25	6.0	10	2.1	28	3.6	26	6.7	116	6.0
	500	26	5.9	7	1.5	25	6.6	20	4.4	118	12.7
	1500	22	7.6	7	1.5	10	1.2	18	2.6	0	0.0
	5000	13	3.2	3	0.6	0	0.0	7	4.2	0	0.0
ENNG	5.0	290	42.9
	3.0									541	18.5
9AA	80			711	112.7
4NOPD	5.0					561	7.0
4NQO	0.2							186	8.2

 

With activation

Test Group	Dose Level (µg/plate)	Revertant Colony Counts
		TA 1535		TA 1537		TA 1538		TA 98		TA100
		mean	SD	mean	SD	mean	SD	Mean	SD	mean	SD
DMSO	Vehicle control	17	4.6	8	3.8	22	3.2	24	3.1	140	15.3
Test	15									131	12.5
	50	15	3,6	10	1.5	24	2.3	24	5.8	155	20.0
	150	16	2.9	9	1.7	23	3.1	28	9.2	148	8.6
	500	20	1.5	6	0.6	20	7.5	30	4.5	146	21.5
	1500	13	1.7	9	1.1	22	6.0	21	5.5	13	1.2
	5000	11	3.6	0	0.0	0	0.0	0	0.0	0	0.0
2AA	2.0	234	26.6	159	26.2
	1.0									1139	49.1
	0.5					227	17.2	224	25.1

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results
negative without metabolic activation
negative with metabolic activation

OPA was considered to be non-mutagenic with and without metabolic activation under the conditions of this test.

Executive summary:

Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 were treated with the test material using the Ames plate incorporation method at up to seven dose levels, in triplicate, both with and without the addition of a rat liver homogenate metabolising system (10% liver S9 in standard co-factors). The dose range was determined in a preliminary toxicity assay and was 5 to 5000 µg/plate in the first experiment. The experiment was repeated on a separate day using a dose range of 15 to 5000 µg/plate with fresh cultures of the bacterial strains and fresh test material formulations.

The vehicle (acetone) control plates produced counts of revertant colonies within the normal range.

All the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with and without the metabolising systems.

The test material caused a visible reduction in the growth of the bacterial background lawn to all the tester strains both with and without metabolic activation. The first indication of a toxic response was observed at 1500 µg/plate.No significant increases in the frequency of revertant colonies was recorded for any of the bacterial strains with any dose of the test material, either with or without metabolic activation. The test material was considered to be non-mutagenic under the conditions of this test.