3-(1-cyanoethyl)benzoyl chloride

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro cytogenicity / micronucleus study

Remarks:

Type of genotoxicity: other: Clastogenicity and aneugenicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 01 March 2011 to 08 March 2011

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study according to international guideline but not GLP study.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

other: OECD guideline 487 dated 22 July 2010

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: IWGT recommendations

Principles of method if other than guideline:

Micronuclei detected in the cytoplasm of interphase cells, result from either chromosome breakage leading to acentric fragments (clastogenicity) or impairment of the structure and/or function of the mitotic apparatus leading to lagging chromosomes (aneugenicity).

GLP compliance:

no

Type of assay:

in vitro mammalian cell micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

3-(1-cyanoethyl)benzoyl chloride

EC Number:

255-980-4

EC Name:

3-(1-cyanoethyl)benzoyl chloride

Cas Number:

42872-29-7

Molecular formula:

C10H8ClNO

IUPAC Name:

3-(1-cyanoethyl)benzoyl chloride

Details on test material:

Test article identification: CFPPN
Test article batch number: CFPPN 2C4.4
Certificate of analysis: dated 7 October 2010
Conversion factor: No as purity is 95.218% ie, >95%

Method

Species / strain

Species / strain / cell type:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Metabolic activation system:

S9-mix from the liver of Aroclor1254-treated rats

Test concentrations with justification for top dose:

With metabolic activation:
Range of concentrations tested in the culture medium: from 1 to 600 μg/mL.
The evaluated concentrations were 50, 200 and 400 μg/mL.
Without metabolic activation
Range of concentrations tested in the culture medium: from 1 to 600 μg/mL.
The evaluated concentrations were 50, 1000 and 200 μg/mL.

Vehicle / solvent:

Acetonitrile

Controls

Untreated negative controls:

yes

Remarks:

Acetonitrile

Negative solvent / vehicle controls:

yes

Remarks:

Acetonitrile

Positive controls:

yes

Positive control substance:

cyclophosphamide

mitomycin C

Remarks:

Without metabolic activation: Mitomycin C (MITC), 0.01 μg/mL / With metabolic activation: Cyclophosphamide (CP), 1.5 μg/mL

Details on test system and experimental conditions:

L5178Y cells were treated with the substance in 96-well microtiter plates for 3 hours with metabolic activation and for 28 hours without metabolic activation. Without metabolic activation the cells were harvested directly after the treatment time and with metabolic activation 21 hours (recovery time) after the end of the treatment.

Evaluation criteria:

Experiment acceptance criteria
The experiment is considered valid when all the following criteria are fulfilled:
• the population doubling of the negative control is higher or equal to 1.5 but not higher than 2.2;
• the number of micronucleated cells per 1000 cells in the concurrent solvent control is lower than or equal to the acceptable upper value for historical solvent control data (99% percentile of our historical negative control data):
- 9 for 28-hour treatment without S9-mix;
- 11 for 3-hour treatment with S9-mix.
• the positive controls induce a marked increase in the number of micronucleated cells;
• at least three analyzable concentrations are scored;
• the highest concentration evaluated corresponds either to 5000 µg/mL (or 10mM), or to the solubility limit in the solvent, or to the lowest precipitating concentration in the culture medium or to 50% to 60% of cell cytotoxicity.

Test evaluation criteria
A test article is considered to induce a positive response in the in vitro micronucleus test if the two criteria listed below are fulfilled for at least one concentration:
• the test article induces a statistically significant increase in the number of micronucleated cells for a given concentration;
• for this concentration, this value is strictly above positive threshold value (99% percentile of our historical negative control data):
- 9 for 28-hour treatment without S9-mix;
- 11 for 3-hour treatment with S9-mix.
When none of the above criteria are fulfilled, the test article is considered negative.

Results and discussion

Test results

Test results 1

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with

Genotoxicity:

negative

Remarks:

No significant increase in the number of micronucleated cells was observed.

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

The top concentration retained for scoring 400 μg/mL (49% of cytotoxicity) was considered as corresponding to the required cytotoxicity (50% to 60%) and corresponded to the lowest precipitating concentration in the culture medium.

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

without

Genotoxicity:

negative

Remarks:

No significant increase in the number of micronucleated cells was observed.

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

The top concentration retained for scoring 200 μg/mL (50 % of cytotoxicity) corresponded to the required cytotoxicity (50% to 60%). The slides of the concentration above (300 μg/mL with 59% of cytotoxicity) were unreadable due to damaged cells.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Under the experimental conditions of this study, CFPPN was found negative in the exploratory in vitro micronucleus test in mouse lymphoma cells in the presence or absence of metabolic activation.