2-hydroxy-5-octanoylbenzoic acid

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:

No data

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: GLP study performed with equivalent protocol to OECD guideline 471 but no certificate of analysis of the test substance and no data about its purity are reported.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1994

Report date:

1994

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

No certificate of analysis of test substance, no data about purity of test substance

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

-

EC Number:

417-790-1

EC Name:

-

Cas Number:

78418-01-6

Molecular formula:

C15H20O4

IUPAC Name:

2-Hydroxy-5-(1-oxooctyl)benzoic acid

Constituent 2

Reference substance name:

MEXORYL SAB

IUPAC Name:

MEXORYL SAB

Details on test material:

- Name of test material (as cited in study report): Mexoryl SAB
- Physical state: white fibrous powder
- Analytical purity: no data
- Lot/batch No.: OP T 4 E
- Storage condition of test material: at room temperature

Method

Target gene:

Not applicable

Species / strain

Species / strain 1

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not applicable

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1538

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not applicable

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Liver microsomal fraction (S9) from Fischer 344 rats induced with beta-naphthoflavone and sodium phenobarbitone

Test concentrations with justification for top dose:

- range finder experiment: 1.6, 8, 40, 200, 1000 and 5000 µg/plate in both the absence and presence of S9
- first experiment: 1.6, 8, 40, 200 and 1000 µg/plate in both the absence and presence of S9
- second experiment: 0.16, 0.8, 4, 20 and 100 µg/plate without S9; 0.32, 1.6, 8, 40 and 200 µg/plate with S9

Vehicle / solvent:

Vehicle: ethanol

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

Ethanol

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: see table 1

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: none
- Exposure duration: about 65 h

SELECTION AGENT (mutation assays): histidine for Salmonella strains, tryptophane for Escherichia coli

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: appearance of a complete bacterial lawn

Evaluation criteria:

A positive result is indicated by a dose response with statistically significant increases in revertant numbers.

Statistics:

Mean, standard deviations and Dunnett's t-statistic were calculated for each group and bacterial strain.

Results and discussion

Test results

Test results 1

Species / strain:

other: TA 98, TA 100, TA1535, TA 1537, TA 1538 and WP2 uvrA

Metabolic activation:

without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

for 100 µg/plate concentration in some of the strains (reduced background lawn)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Species / strain:

other: TA 98, TA 100, TA1535, TA 1537, TA 1538 and WP2 uvrA

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

for 200 µg/plate concentration in some of the strains (reduced background lawn)

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: TA 98 and WP2 uvrA were used with and without S9 during 24 h; Mexoryl SAB had slight effects on bacterial growth for 1000 and 5000 µg/plate concentrations. Therefore the strains were exposed up to 1000 µg/plate in the main test.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 2: results of the first experiment

Strain	Compound	Dose level (µg/plate)	S9 mix	Mean revertant colony counts	SD	Individual revertant colony counts
TA 1535	Ethanol           Sodium azide   Ethanol           2-aminoanthracene	1.6 8 40 200 1000 1     1.6 8 40 200 1000 2	- - - - - - -   + + + + + + +	7.3 7.3 13.3 10.3 - - 96   13.3 15.3 14.3 19.0 15.7 - 48	1.53 3.79 2.31 1.15 - - 10   3.06 4.51 1.53 6.24 5.03 - 11.36	6, 9, 7 9, 3, 10 16, 12, 12 9, 11, 11 a b 96, 86, 106   10, 16, 14 20, 15, 11 13, 16, 14 26, 17, 14 21, 11, 15 a 40, 43, 61
TA 1537	Ethanol           9-aminoacridine   Ethanol           2-aminoanthracene	1.6 8 40 200 1000 50     1.6 8 40 200 1000 2	- - - - - - -   + + + + + + +	3.0 5.0 2.0 3.7 - - 295.0   9.3 4.0 5.7 6.0 8.3 - 76.3	1.73 1.00 1.00 1.15 - - 35.04   6.66 4.36 0.58 1.00 0.58 - 8.50	5, 2, 2 4, 5, 6 1, 3, 2 5, 3, 3 a b 331, 261, 293   6, 17, 5 9, 2, 1 6, 5, 6 7, 5, 6 8, 8, 9 a 86, 70, 73
TA 1538	Ethanol           2-Nitrofluorene   Ethanol           2-aminoanthracene	1.6 8 40 200 1000 0.5     1.6 8 40 200 1000 2	- - - - - - -   + + + + + + +	10.7 10.7 15.7 9.3 - - 232.7   24.3 19.3 24.3 26.7 19.3 - 1305.3	2.08 2.08 7.02 2.52 - - 34.24   7.02 0.58 4.51 3.79 7.37 - 137.38	13, 9, 10 9, 10, 13 15, 9, 23 7, 9, 12 a b 269, 201, 228   31, 17, 25 19, 20, 19 29, 24, 20 31, 24, 25 11, 25, 22 a 1393, 1376, 1147
TA 98	Ethanol           2-Nitrofluorene   Ethanol           2-aminoanthracene	1.6 8 40 200 1000 0.5     1.6 8 40 200 1000 2	- - - - - - -   + + + + + + +	19.0 17.7 12.0 21.7 - - 249.0   25.0 19.0 20.0 22.0 25.3 - 923.3	2.65 4.62 1.73 7.57 - - 3.61   7.55 1.73 2.65 5.57 9.29 - 50.90	18, 22, 17 15, 23, 15 14, 11, 11 25, 13, 27 a b 250, 252, 245   26, 32, 17 17, 20, 20 18, 23, 19 17, 21, 28 33, 15, 28 b 937, 966, 867
TA 100	Ethanol           Sodium azide   Ethanol           2-aminoanthracene	1.6 8 40 200 1000 1     1.6 8 40 200 1000 2	- - - - - - -   + + + + + + +	68.7 67.7 74.0 72.3 - - 466.0   83.3 82.0 85.7 77.0 61.3 - 951.3	13.32 8.50 6.00 12.22 - - 60.51   12.22 14.00 8.39 9.17 4.62 - 122.68	72, 54, 80 59, 76, 68 74, 68, 80 59, 83, 75 a a 441, 422, 535   86, 70, 94 66, 88, 92 91, 90, 76 69, 75, 87 64, 64, 56 a, b, b 861, 1091, 902
WP2 uvrA	Ethanol             4 -nitroquinoline-1-oxide   Ethanol           2-aminoanthracene	1.6 8 40 200 1000     1     1.6 8 40 200 1000 2	- - - - - - -       + + + + + + +	15.0 14.7 13.3 16.3 - -     617.7   17.0 13.7 19.3 14.7 - - 94.7	3.46 4.51 4.93 1.15 - -     32.65   1.73 5.77 5.51 2.31 - - 11.55	13, 13, 19 15, 10, 19 10, 11, 19 15, 17, 17 b b     580, 638, 635   18, 18, 15 7, 17, 17 23, 22, 13 16, 16, 12 a b 108, 88, 88

a = reduced background lawn

b = complete loss of lawn

Table 3: results of the second experiment

Strain	Compound	Dose level (µg/plate)	S9 mix	Mean revertant colony counts	SD	Individual revertant colony counts
TA 1535	Ethanol           Sodium azide   Ethanol           2-aminoanthracene	0.16 0.8 4 20 100 1     0.32 1.6 8 40 200 2	- - - - - - -   + + + + + + +	12.7 10.7 9.3 9.0 9.7 - 252.0   10.7 14.3 15.0 11.3 15.7 - 106.0	4.16 2.52 3.06 1.73 4.73 0.00 3.61   4.93 4.51 6.56 4.04 3.21 - 7.81	14, 8, 16 8, 11, 13 6, 10, 12 7, 10, 10 15, 8, 6 a 251, 256, 249   5, 14, 13 19, 14, 10 14, 22, 9 15, 7, 12 12, 17, 18 a 102, 101, 115
TA 1537	Ethanol           9-aminoacridine   Ethanol           2-aminoanthracene	0.16 0.8 4 20 100 50     0.32 1.6 8 40 200 2	- - - - - - -   + + + + + + +	4.7 2.0 2.0 3.7 4.3 - 852.7   11.3 6.3 6.7 6.0 8.7 - 101.3	2.52 1.00 1.00 2.08 1.53 0.00 8.14   1.53 0.58 0.58 1.73 1.53 - 22.19	2, 7, 5 1, 2, 3 1, 2, 3 2,3, 6 3, 4, 6 a 849, 847, 862   13, 10, 11 6, 7, 6 6, 7, 7 8, 5, 5 9, 10, 7 a 126, 83, 95
TA 1538	Ethanol           2-Nitrofluorene   Ethanol           2-aminoanthracene	0.16 0.8 4 20 100 0.5     0.32 1.6 8 40 200 2	- - - - - - -   + + + + + + +	17.7 18.3 18.3 17.3 16.7 - 90.0   22.3 21.0 22.3 25.3 26.0 20.0 1225.0	1.53 4.73 1.53 3.06 3.21 0.00 10.82   6.35 5.20 3.51 6.66 3.46 2.65 32.60	16, 18, 19 22, 20, 13 18, 17, 20 18, 14, 20 19, 18, 13 a 102, 87, 81   15, 26, 26 27, 18, 18 26, 19, 22 18, 27, 31 24, 24, 30 18, 23, 19 1191, 1228, 1256
TA 98	Ethanol           2-Nitrofluorene   Ethanol           2-aminoanthracene	0.16 0.8 4 20 100 0.5     0.32 1.6 8 40 200 2	- - - - - - -   + + + + + + +	14.0 15.7 12.0 10.0 14.3 14.3 81.0   24.0 22.0 23.3 21.0 22.3 17.3 1111.7	1.00 4.16 1.73 1.00 1.15 2.89 10.15   3.46 2.00 5.51 4.36 2.89 0.58 31.79	15, 14, 13 19, 17, 11 10, 13, 13 10, 9, 11 15, 13, 15 16, 11, 16 92, 79, 72   22, 28, 22 22, 24, 20 29, 23, 18 24, 23, 16 24, 19, 24 17, 18, 17 1098, 1148, 1089
TA 100	Ethanol           Sodium azide   Ethanol           2-aminoanthracene	0.16 0.8 4 20 100 1     0.32 1.6 8 40 200 2	- - - - - - -   + + + + + + +	67.0 70.0 76.0 70.7 67.0   421.0   81.7 80.7 89.0 81.0 73.0 66.0 1211.7	2.65 7.55 7.00 6.66 7.94 0.00 5.57   7.09 8.39 11.36 11.36 6.93 5.57 33.08	64, 68, 69 69, 63, 78 68, 79, 81 78, 65, 69 64, 61, 76 a 415, 426, 422   83, 88, 74 85, 71, 86 76, 94, 97 89, 86, 68 81, 69, 69 72, 65, 61 1249, 1186, 1200
WP2 uvrA	Ethanol           4 -nitroquinoline-1-oxide   Ethanol           2-aminoanthracene	0.16 0.8 4 20 100   1     0.32 1.6 8 40 200 2	- - - - - -   -   + + + + + + +	15.3 14.7 14.0 15.7 15.3 -   832.3   18.7 14.3 16.7 16.0 16.3 - 127.7	1.53 4.62 3.46 3.21 4.04 -   55.82   6.66 4.73 1.15 2.65 0.58 - 4.62	15, 14, 17 20, 12, 12 16, 10, 16 17, 18, 12 13, 20, 13 a   861, 768, 868   26, 17, 13 18, 9, 16 16, 16, 18 13, 18, 17 16, 17, 16 a 133, 125, 125

a = reduced background lawn

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Under these test conditions, Mexoryl SAB was found not to be mutagenic in TA 1535, TA 1537, TA 1538, TA 100, TA 98 and WP2 uvrA with and without metabolic activation.

Executive summary:

In a reverse gene mutation assay in bacteria, performed similarly to the OECD guideline 471, in compliance with GLP, strains of S. typhimurium (TA 1535, TA 1537, TA 1538, TA 100 and TA 98) and one Escherichia coli strain (WP2 uvrA) were exposed to Mexoryl SAB at concentrations of 1.6, 8, 40, 200, 1000 and 5000 µg/plate in both the absence and presence of S9 metabolic activation according to the direct plate incorporation method for a preliminary cytotoxicity test. In the main test, two experiments were performed at the following concentrations:

- first experiment: 1.6, 8, 40, 200 and 1000 µg/plate in both the absence and presence of S9

- second experiment: 0.16, 0.8, 4, 20 and 100 µg/plate without S9; 0.32, 1.6, 8, 40 and 200 µg/plate with S9.

These experiments were performed according to the direct plate incorporation method.

The positive controls induced the appropriate responses in the corresponding strains. Mexoryl SAB showed no substantial increases in revertant colony numbers over control count obtained with any of the tester strains at any concentrations in either presence or absence of S9 mix.

This study is classified as acceptable and satisfies the requirement for bacterial reverse gene mutation endpoint.

Under the test conditions, Mexoryl SAB is not considered as mutagenic in this bacterial system according to the criteria of the Annex VI to the Directive 67/548/EEC and CLP Regulation (EC) N° (1272-2008).