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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information
The substance 3-Phenoxybenzylalcohol was found to be non mutagenic in in-vitro study.
Link to relevant study records
Reference
Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: Data is from online Chemical Repository Database
Qualifier:
according to
Guideline:
other:
Principles of method if other than guideline:
Salmonella/microsomal assay to investigate the mutagenic potentlaI of a test compound 3-Phenoxybenzyl alcohol (3-PBA)
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay
Species / strain / cell type:
S. typhimurium, other: TA1535, TA100, TA1537, TA1538 and TA98
Details on mammalian cell type (if applicable):
- Type and identity of media: Spizzizen's minimal medium ,Noble's agar,Nutrient broth
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S-9 from Sprague-Dawley rats
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
other: N-methyl- N -nitro-N-nitrosoguanidine,2-Aminoanthracene,2-Aminofluorene, 6-Aminochrysene, Aflatoxin B1
Details on test system and experimental conditions:
METHOD OF APPLICATION :in agar (plate incorporation)

Evaluation criteria:
significant increase in the number of revertant mutant colonies when compared to the spontaneous background rate which occurs in the control cultures
Statistics:
Linear regression analysis to determine strength of dose response
Species / strain:
S. typhimurium, other: TA1535, TA100, TA1537, TA1538 and TA98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
not specified
Untreated negative controls validity:
not specified
Positive controls validity:
not specified
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

RESULTS OF SALMONELLA/MICROSOMAL ASSAY WITHOUT METABOLIC ACTIVATION ON 3-PBA

compound

conc

units/

plate

REVERTANTS PER PLATE OF BACTERIAL TESTER STRAIN

TA1535           TA1537             TA1538              TA100              TA98

bacteria only

(negative control)

 

100.0000 µL

    7.0

 ± 2.0

    5.3

 ± 1.2

    9.7

 ± 1.5

    126.7

 ±   10.4

    15.3

 ±   5.5

DMSO (SOLVENT CONTROL)

100.0000 µL

    9.0

 ± 2.6

    5.3

 ± 1.5

    8.7

 ± 3.5

    139.0

 ±   24.2

    16.0

 ±   1.0

MNNG

(POSITIVE CONTROL)

5.0000 µg

    813.7

  ± 66.2

      NT

   NT

    855.3

 ± 203.3

    NT

DMSO (POSITIVE CONTROL)

100.0000 µg

NT

    463.3

 ±   62.4

   NT

    NT

    NT

DMSO (POSITIVE CONTROL)

5.000 µg

NT

      NT

    264.3

 ± 110.6

    NT

     271.3

 ±    40.4

3-PBA

.0050 µL

    11.3

 ±   2.1

    5.0

 ± 2.0

    4.0

 ± 1.0

    229.3

 ±   29.3

    15.0

 ±   3.5

0.5 % SOLUTION (v/v) IN DMSO

.0100 µL

    16.3

 ±   3.2

    3.7

 ± 2.1

    4.0

 ± 1.0

    187.7

 ±   18.6

    15.3

 ±   4.5

.050 0 µL

    16.7

 ±   2.5

    6.0

 ± 2.6

    6.7

 ± 3.1

    201.7

 ±   26.8

    16.3

 ±   2.5

.1000 µL

     13.7

 ±    1.5

    6.3

 ±   .6

    3.3

 ± 2.5

    189.7

 ±     7.8

     17.7

 ±    2.3

.5000 µL

    10.3

 ±   1.5

    1.7

 ± 0.6

    6.0

 ±   .0

    155.3

 ±   21.4

     17.7

 ±    2.1

NT: not tested

 

RESULTS OF SALMONELLA/MICROSOMAL ASSAY WITH METABOLIC ACTIVATION ON 3-PBA

compound

conc

units/

plate

Revertants per plate of bacterial tester strain

TA1535           TA1537             TA1538              TA100              TA98

Bacteria only

(negative control)

 

100.0000 µL

    7.0

 ± 2.0

    5.3

 ± 1.2

    9.7

 ± 1.5

    126.7

 ±   10.4

    15.3

 ±   5.5

S-9 FRACTION(NEGATIVE CONTRL)

100.0000 µL

    8.3

 ±  .6

    5.3

 ± 0.6

    9.3

 ± 2.3

    131.7

 ±   41.5

    21.7

 ±   3.1

DMSO( SOLVENT CONTRL)

100.0000 µL

       9.7

  ±  2.5

      6.3

 ±  1.2

   11.3

 ±  3.8

    126.3

 ±   38.1

    17.3

±    8.4

2-AA(POSITIVE CONTROL)

5.0000 µg

       74.0

  ±  10.1

     NT

    NT

    NT

    NT

6-AC(POSITIVE CONTROL)

1.000 µg

NT

      51.3

 ±    7.1

    NT

    NT

    NT

2-AF(POSITIVE CONTROL)

2.000 µg

    NT

    NT

    256.7

 ±   64.5

    NT

    NT

AFLTOXIN

(POSITIVE CONTROL)

1.000 µg

    NT

    NT

    NT

    656.3

 ± 203.9

    307.0

 ±   66.8

3-PBA

0.5 % SOLUTION

(V/V) IN DMSO

.0050 0 µL

    14.7

 ±   2.3

    13.3

 ±   2.1

    18.7

 ±   5.0

    280.3

 ±   18.6

    18.7

 ±   5.0

.01000 µL

     8.3

 ±  1.5

    16.7

 ±   4.0

    13.0

 ±   1.0

    245.7

 ±     4.5

     22.3

 ±    8.5

.05000 µL

     7.0

 ±  0.0

    12.0

 ±   2.6

    15.3

 ±   1.5

    243.7

 ±   16.8

     23.3

 ±    3.5

.1000 µL

   12.0

 ±  4.6

     13.3

 ±    5.5

    11.7

 ±   2.5

    216.0

 ±   10.4

    20.0

 ±   7.5

.5000 µL

  12.3

 ± 4.2

    10.0

 ±  1.0

      5.7

 ±   2.1

   175.0

 ±  39.9

    12.0

 ±   2.6

NT: not tested

Conclusions:
Interpretation of results (migrated information):
negative

The genetic toxicity test for 3-Phenoxybenzylalcohol (CAS No: 13826-35-2) on salmonella strains TA1535, TA100, TA1537, TA1538 and TA98 was found to be negative with and without activation.
Executive summary:

Genetic toxicity test was performed by using 3-Phenoxybenzylalcohol onsalmonella strainsTA1535, TA100, TA1537, TA1538 and TA98 with and without metabolic activation.Liver S-9 preparations were done from Sprague-Dawley male rats pretreated with Aroclor 1254 for the non-specific induction of hepatic enzymes served as the activation system. S-9 preparation was used at a final concentration of 100 µl per ml of S-9 mix. Dimethyl sulphoxide (DMSO) was the solvent used. Spizzizen's minimal medium, Noble's agar, Nutrient broth were the medium used in these experiments.

 

The test was performed according to the method of Ames et al.For the assay, the following were added, in order to 16 x 125 mm screw-capped tubes:

1.2 ml molten agar (0.6% Noble's agar supplemented with 0.6% NaCl , 0.5 mM biotin, and 0.5 mM histidine).

2.0.1 ml of bacterial culture.

3.0.1 ml of test agent in the appropriate solvent.

Tubes were mixed quickly but gently and the contents poured over a base plate of Spizzizen's minimal medium. Plates were rotated to distribute the overlay agar, allowed to harden and incubated at 370C for 48 hr at which time the number of mutant colonies were determined.

In case of metabolic activation with S-9, all procedure was same except that 0.5 ml of S-9 mix was added to the overlay agar immediately before it was mixed and poured. Plates were incubated.

 

Each agent was tested at five concentrations against all five strains of bacteria.

The following controls were included in each experiment:

Bacteria only for spontaneous reversion (Negative control), Bacteria and solvent for solvent control.Chemical used as positive control:Without metabolic activation areN-methyl- N -nitro-N-nitrosoguanidine, 2-Nitrofluorene, 9-Aminoacridineandwith metabolic activation:2-Aminoanthracene, 2-Aminofluorene, 6-Aminochrysene, Aflatoxin B1.   

 

The test agent did not induce a significant increase in the number of point mutations inSalmonella typhimuriumstrains in the absence and in the presence of the activating system for strains TA1535, TA1537, TA1538, TA100, and TA98.

 

So from the above test procedure it was concluded thatthe genetic toxicity test for 3-Phenoxybenzylalcohol (CAS No: 13826-35-2) onsalmonellastrains TA1535, TA100, TA1537, TA1538 and TA98 was found to be negative with and without activation.

 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

Gene toxicity in vitro:

Various reliable studies were reviewed to determine the mutation potential of the test compound 3 -Phenoxybenzylalcohol (CAS no 13826 -35 -2). These are summarized as below:

Genetic toxicity test was performed by using 3-Phenoxybenzylalcohol on salmonella strainsTA1535, TA100, TA1537, TA1538 and TA98 with and without metabolic activation.Liver S-9 preparations were done from Sprague-Dawley male rats pretreated with Aroclor 1254 for the non-specific induction of hepatic enzymes served as the activation system. S-9 preparation was used at a final concentration of 100 µl per ml of S-9 mix. Dimethyl sulphoxide (DMSO) was the solvent used. Spizzizen's minimal medium, Noble's agar, Nutrient broth were the medium used in these experiments.

 

The test was performed according to the method of Ames et al.For the assay, the following were added, in order to 16 x 125 mm screw-capped tubes:

1.2 ml molten agar (0.6% Noble's agar supplemented with 0.6% NaCl , 0.5 mM biotin, and 0.5 mM histidine).

2.0.1 ml of bacterial culture.

3.0.1 ml of test agent in the appropriate solvent.

Tubes were mixed quickly but gently and the contents poured over a base plate of Spizzizen's minimal medium. Plates were rotated to distribute the overlay agar, allowed to harden and incubated at 370C for 48 hr at which time the number of mutant colonies were determined.

In case of metabolic activation with S-9, all procedure was same except that 0.5 ml of S-9 mix was added to the overlay agar immediately before it was mixed and poured. Plates were incubated.

 

Each agent was tested at five concentrations against all five strains of bacteria.

The following controls were included in each experiment:

Bacteria only for spontaneous reversion (Negative control), Bacteria and solvent for solvent control.Chemical used as positive control:Without metabolic activation areN-methyl- N -nitro-N-nitrosoguanidine, 2-Nitrofluorene, 9-Aminoacridineandwith metabolic activation:2-Aminoanthracene, 2-Aminofluorene, 6-Aminochrysene, Aflatoxin B1.   

 

The test agent did not induce a significant increase in the number of point mutations in Salmonella typhimuriumstrains in the absence and in the presence of the activating system for strains TA1535, TA1537, TA1538, TA100, and TA98.

 

So from the above test procedure it was concluded thatthe genetic toxicity test for 3-Phenoxybenzylalcohol (CAS No: 13826-35-2) on salmonellastrains TA1535, TA100, TA1537, TA1538 and TA98 was found to be negative with and without activation.

 

In another Salmonella/ microsomal assay performed (Ethyl Corporation, 1993), the mutagenic potential of a test compound 3-Phenoxybenzyl Alcohol was investigated. The test was performed at five concentrations ranging from 0.0100. 0.0500, 0.1000, 0.5000 or 1.0000 µL/plate in triplicate. The plates were scored after 48 hrs exposure period. Appropriate solvent, negative and positive controls were incorporated in the study. The mutagenic potential of a test chemical was determined by its ability to induce a significant increase in the number of revertant mutant colonies when compared to the spontaneous background rate which occurs in the control cultures. The test compound3-Phenoxybenzyl Alcohol did not induce a significant the number of point mutations inSalmonellastrains TA1535, TA100, TA1537, TA1538 and TA98 in the presence or absence of the activating system S9.

Gene toxicity in vivo:

Dominant Lethal Assay was performed (Ethyl Corporation, 1993) to evaluate the mutagenic nature (in vivo) of the test compound 3-Phenoxybenzyl alcohol on Sprague Dawley rats.

 

Three groups of male rats, 10 per group, were given five intragastric treatments with the test agent at levels of LD5, LD0.5and LD0.05from Monday through Friday. The following week each male was then mated with 2 females from Monday through Friday. On Friday, the females were removed from the cage and the male was allowed to rest for 2 days. On the subsequent Monday, the male was mated with 2 new females and the process repeated until each male had been mated for 7 weeks with 2 new females per week. Fourteen days from the mid-week of mating, the females were sacrificed by CO2 asphyxiation and the abdominal cavity was exposed. The membrane was removed from each ovary and the corpora lutea for each ovary were counted and recorded separately. In addition, both uterine horns were examined and fetal deaths and total implantations were determined and recorded separately for each horn.

 

The test compound 3-PBA appeared to exhibit minimal activity in number of total implantations, preimplantation losses, and number of live implants per pregnant female. However, the effect on total or live implants per pregnant female occurred at a single dose level at week 3 and 7. The variation in corpora lutea counts does not reflect any biological activity of the doses tested but rather is a variation in the female animals. The increase seen in pre-implantation losses occured at a single dose at week 1 only. No statistically significant dose response was observed.

 

The data suggest that the test compound exhibits little or no mutagenic activity in the dominant lethal test.


Justification for selection of genetic toxicity endpoint
The genetic toxicity test for 3-Phenoxybenzylalcohol (CAS No: 13826-35-2) on salmonella strains TA1535, TA100, TA1537, TA1538 and TA98 was found to be negative with and without activation.

Justification for classification or non-classification

The substance 3-Phenoxybenzylalcohol was found to give negative effect and hence classified as non-mutagenic.