3-chloropropionyl chloride

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

In an in vitro mammalian chromosome aberration test the test substance was found not to be clastogenic in human lymphocytes. In an Ames test, the test substance proved mutagenic with and without metabolic activation. In an E.coli SOS chromotest without metabolic activation the test substance was also tested positve for genetic toxicty. A structural analogue of the test substance also proved to be mutagenic using Salmonella typhimurium (TA 100) without metabolic activation.

Link to relevant study records

Reference

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Species / strain / cell type:

lymphocytes: human (from heparinized human venous blood)

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced rat S-9 mix

Test concentrations with justification for top dose:

5, 10, 20, 40 and 60 µg/mL (without activation system), 60, 125, 250, 500, 750 µg/mL (with activation system)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: culture medium

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

0.1 µg/mL

Positive control substance:

mitomycin C

Remarks:

without S-9 mix

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Remarks:

5 µg/mL

Positive control substance:

cyclophosphamide

Remarks:

with S-9 mix

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: 48 hours
- Exposure duration: Without metabolic activation: 24 hours; With metabolic activattion: 24 hours (After about 3 hours of incubation at 37°C, cells were washed twice with unsupplemented culture medium and then re-incubated in complete culture medium for further 21 hours).
- Fixation time (start of exposure up to fixation or harvest of cells): 72 hours

SPINDLE INHIBITOR: colcemid

STAIN: Giemsa/Titrisol solution

NUMBER OF REPLICATIONS: 2

NUMBER OF CELLS EVALUATED: 100 metaphases

DETERMINATION OF CYTOTOXICITY: mitotic index

Evaluation criteria:

As a rule, 100 metaphases of each culture for the test substance, negative and solvent controls or 50 cells of each culture for the positive controls were analyzed for chromosome aberrations. A test substance was positive in this test if it led to a biological significant increase in the number of aberrant metaphases incl. and excl. gaps when compared to the untreated control or to the historical control.

Statistics:

The statistical evaluation of the data was carried out using the MUCHAN program system (BASF SE).
For each group the proportion of metaphases with aberrations was calculated.
A comparison of each dose group with the negative control group was carried out using Fisher's exact test for the hypothesis of equal proportions. This test was Bonferoni-Holm corrected over the dose groups separately for each time point and was performed one-sided.

Species / strain:

lymphocytes: human (from heparinized human venous blood)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

According to the results of the present study, the test substance did not cause a biological significant increase in the number of aberrant metaphases incl. and excl. gaps when compared to the untreated control or to the historical control data (2.0 - 12.0% incl. gaps or 0.5 - 4.0% excl. gaps).

No differences regarding aneuploidies (hyperploid metaphases) and polyploidies between the varous dose groups and the negative control were observed.

Remarks on result:

other: all strains/cell types tested

Additional information

In an in vitro mammalian chromosome aberration test performed similar to OECD guideline 473 (1991; RL1), the test substance was tested for the ability to induce chromosomal aberrations in human lymphocytes following exposure in the presence and absence of a metabolizing system (Aroclor 1254 induced rat S-9 mix). In the absence of S-9 mix the test concentrations were 5, 10, 20, 40 and 60 µg/mL and with S-9 mix 60, 125, 250, 500, 750 µg/mL. For control purposes, negative controls (untreated) and positive controls: mitomycin C and cyclophosphamide were tested. The test substance did not cause an increase in the number of aberrant metaphases incl. and excl. gaps either without S-9 mix or after adding a metabolizing system. Thus, under the experimental conditions chosen here the test substance has no chromosome-damaging (clastogenic) effect in vitro using human lymphocytes.

An Ames test (OECD TG 471, GLP) is available in which the tester strains Salmonella typhimurium TA 1535, TA 1537, TA1538, TA 98 and TA 100 were used to examine the mutagenic potential of the test substance up to concentrations of 5000 µg/plate in a standard plate test with and without metabolic activation (1991; RL2). In the strains TA 1535, TA 1537 and TA 100 a concentration-dependent increase in revertants both with and without metabolic activation was observed. For TA 1535 the increase was already observed at the lowest concentration (10 µg/plate). For the other 2 strains the lowest effective concentration was 100 µg/plate. The test substance thus proved to be mutagenic in this test system.

The test substance was also tested, without metabolic activation, in the E.coli SOS chromotest (Szegedi 1998, abstract only, RL4). The experiments were carried out with a BIOSCREEN Analyzing System using the BIOSOS program (LABSYSTENS Ltd.). The test substance tested positve for genetic toxicty under the experimental conditions chosen here.

As a structural analogue of the test substance: 3-chloropropionic acid, CAS no. 107 -94 -8 (hydrolysis product of 3 -chloropropionyl chloride, CAS no. 625 -36 -5) was tested in a reverse mutation assay (Ames test) using strains of Salmonella typhimurium (TA 100 and 1535) in the presence and absence of metabolic activation (1987; RL2). Tested concentrations were 50, 100, 250, 500 and 1000 µg/plate. Only the results of the TA 100 strain without metabolic activation were presented in the paper. The test substance was mutagenic, yielding 1000 revertants/250 µg.

Justification for classification or non-classification

Based on the available information, classification for genetic toxicity is not possible in accordance with EU Classification, Labeling and Packaging of Substances and Mixtures (CLP) Regulation No. 1272/2008.