Sodium cumenesulphonate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

There are conclusive but not suffcient data for the classification of substance Sodium cumenesulphonate w ith regard to mutagenicity/genetic toxicity. It is concluded that the substance Sodium cumenesulphonate does not meet the criteria to be classified for human health hazards for Mutagenicity-Genetic Toxicity

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

CAS Number: 28348-53-0
Identity: Cumene sulfonic acid, sodium salt; Na cumene sulfonate
Purity: 40% active ingredient
Remarks: Substances tested are aqueous solutions; % purity equates to chemical content

Target gene:

Histidine independent mutant colonies of Salmonella typhimurium

Species / strain / cell type:

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

Species / strain / cell type:

S. typhimurium TA 1538

Metabolic activation:

with and without

Metabolic activation system:

- Species and cell type: Rat (Sprague Dawley strain), male, liver - Quantity: 5 % S9 mix induced with Aroclor 1254

Test concentrations with justification for top dose:

3.2, 16, 80, 400 and 2000 μg active ingredient

Vehicle / solvent:

vehicle (water), all strains.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: Positive controls (no metabolic activation):sodium-azide for TA 1535 and TA 100. 4-nitro-o-phenylenediamine for TA 1537, TA 1538 and TA 98. Positive control (with metabolic activation): 2-aminoanthracene, all strains

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)


DURATION
- Preincubation period: no data
- Exposure duration: 2 days
- Expression time (cells in growth medium):
- Selection time (if incubation with a selection agent):
- Fixation time (start of exposure up to fixation or harvest of cells):


SELECTION AGENT (mutation assays):
SPINDLE INHIBITOR (cytogenetic assays):
STAIN (for cytogenetic assays):


NUMBER OF REPLICATIONS: triplicates and entire trial repeated once


NUMBER OF CELLS EVALUATED: histidine independent colonies


DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other:


OTHER EXAMINATIONS:
- Determination of polyploidy:
- Determination of endoreplication:

Evaluation criteria:

The assay was considered valid if the number of spontaneous revertant colonies in vehicle control plates falls within the normal range and the positive control chemicals induce significant increases in the number of mutagen-induced revertant colonies compared to vehicle control. It was judged to be positive if there was a concentration-related increase over the range tested and/or a reproducible increase at one or more concentrations in the number of revertant colonies per plate in at least one strain with or without the metabolic system. In addition, it was judged to have a toxic effect (antibacterial effect) when a clearing or diminution of background lawn, the appearance of micro-colonies, and/or thedecrease more than 50% in the number of colonies compared to that of vehicle control was observed.

Species / strain:

other: Salmonella typihimurium TA 1535; TA 100; TA 1537; TA 1538; TA98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Conclusions:

Interpretation of results :negative

In the presence and absence of metabolic system, no mutation in the Salmonella typhimurium (strains TA 98, TA 100, TA 1535 and TA 1537) occurred with sodium cumenesulphonate

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

other: published data

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

equivalent or similar to guideline

Guideline:

EPA OPPTS 870.5375 - In vitro Mammalian Chromosome Aberration Test

GLP compliance:

yes

Type of assay:

other: in vitro mammalian chromosome aberration test

Specific details on test material used for the study:

CAS Number: 1300-72-7
Identity: Xylene sulfonic acid, sodium salt
Purity: 65%
Remarks: purity 65% (11.5% ortho, 38% meta and 15.5% para)

Species / strain / cell type:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced male Sprague-Dawley rat liver S9

Test concentrations with justification for top dose:

2513, 3750 and 5000 micrograms active ingredient per milliliter

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:

cyclophosphamide

Remarks:

with S9 and mitomycin-C without S9

Details on test system and experimental conditions:

Without S9, cells were incubated in McCoy's 5A medium with the test substance for 18 hours. colcemid was then added and incubation continued for additional 2 hours. The cells were harvested by mitotic shake-off, fixed and stained with Giemsa. The cells with S9 activation were treated with the test substance for 2 hours at which time the treatment medium was removed and the cells were incubated for 10 hours in fresh medium with Colcemid present for the last 2 hours. Harvesting was the same.

Evaluation criteria:

Cells were selected for scoring on the basis of good morphology and completeness of karyotype (21 +/- 2 chromosomes). All slides were scored blind and those from a single test were read by the same person. Two hundred first-division metaphase cells were scored at each test level. Aberrations were classified as "simple" (breaks and terminal deletions), "complex" (rearrangements and translocations) and "other" (pulverized cells, despiralized chromosomes, and cells containing 10 or more aberrations).

Statistics:

Analyses were conducted on both the dose response curve and individual dose points. For a single trial, a significant (P

Species / strain:

Chinese hamster Ovary (CHO)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Remarks:

but tested up to limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: strain/cell type: CHO cells

Conclusions:

Interpretation of results :negative
Test substance did not induce chromosomal aberrations at the concentration range tested in CHO cells under the present experimental conditions.

Reference 3

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

(Q)SAR

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification

Justification for type of information:

QSAR prediction:QSAR method for chemicals properties assessment. Relevant for in vitro (Ames test) mutagenicity endpoints.

Qualifier:

according to guideline

Guideline:

other: ToxTree: Benigni/Bossa rules for carcinogenicity and mutagenicity

Principles of method if other than guideline:

This profiler is based on the Mutagenicity/Carcinogenicity module of the software Toxtree. It works as a decision tree for estimating in vitro (Ames test) mutagenicity, based on a list of 30 structural alerts (SAs). The SAs for mutagenicity are molecular functional groups or substructures known to be linked to the mutagenic activity of chemicals. As one or more SAs embedded in a molecular structure are recognised, the system flags the potential mutagenicity of the chemical. The present list of SAs is a subset of the original Toxtree list, obtained by eliminating the SAs for nongenotoxic carcinogenicity.

GLP compliance:

no

Remarks:

not applicable. QSAR model in vitro (Ames test) mutagenicity, based on a list of 30 structural alerts (SAs) relevant for in vitro (Ames test) mutagenicity endpoints.

Type of assay:

other: QSAR model

Target gene:

This profiler is based on the Mutagenicity/Carcinogenicity module of the software Toxtree. It works as a decision tree for estimating in vitro (Ames test) mutagenicity, based on a list of 30 structural alerts (SAs).

Species / strain / cell type:

S. typhimurium TA 100

Test concentrations with justification for top dose:

QSAR model in vitro (Ames test) mutagenicity, based on a list of 30 structural alerts (SAs) relevant for in vitro (Ames test) mutagenicity endpoints.

Untreated negative controls:

other: QSAR model

Negative solvent / vehicle controls:

other: QSAR model

True negative controls:

other: QSAR model

Positive controls:

other: QSAR model

Details on test system and experimental conditions:

This profiler is based on the Mutagenicity/Carcinogenicity module of the software Toxtree.

Evaluation criteria:

This profiler is based on the Mutagenicity/Carcinogenicity module of the software Toxtree. It works as a decision tree for estimating in vitro (Ames test) mutagenicity, based on a list of 30 structural alerts (SAs). The SAs for mutagenicity are molecular functional groups or substructures known to be linked to the mutagenic activity of chemicals. As one or more SAs embedded in a molecular structure are recognised, the system flags the potential mutagenicity of the chemical. The present list of SAs is a subset of the original Toxtree list, obtained by eliminating the SAs for nongenotoxic carcinogenicity.

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

other: QSAR model

Untreated negative controls validity:

other: QSAR model

Additional information on results:

Benigni/Bossa rules for carcinogenicity and mutagenicity:
- Structural Alert for genotoxic carcinogenicity NO
- Potential S. typhiunium TA100 mutagen based on QSAR NO
- Negative for genotoxic carcinogenicity YES

1.6. Profiling results:

DNA binding by OECD

No alert found

Est rogen Receptor Binding

Non binder, without OH or NH2 group

OECD HPV Chemical Categories

Hydrotrope surfactants

Linear alkylbenzene sulfonates

Protein binding by OECD

No alert found

Protein binding potency

Not possible to classify according to these rules (GSH)

Superfragments

No superfragment

Toxic hazard classification by Cramer (original)

High (Class III)

US-EPA New Chemical Categories

Not categorized

Conclusions:

Interpretation of results :negative
No alert found.The query structure is not recognized among the in vitro mutagenicity (Ames test) alerts by ISS.
The query structure is not recognized among the in vitro mutagenicity (Ames test) alerts by ISS and therefore sodium cumenesulphonate does not cause in vitro mutagenicity (Ames test)

Executive summary:

The query structure is not recognized among the in vitro mutagenicity (Ames test) alerts by ISS and does not cause in vitro mutagenicity (Ames test).

No mutagenic activity in the (Q)SAR study, In vitro mutagenicity (Ames test) alerts by ISS for  sodium cumenesulphonate

and does not cause in vitro mutagenicity (Ames test) .This QSAR method is Relevant for in vitro (Ames test) mutagenicity endpoints.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Description of key information

There are conclusive but not suffcient data for the classification of substance Sodium cumenesulphonate with regard to mutagenicity/genetic toxicity. It is concluded that the substance Sodium cumenesulphonate does not meet the criteria to be classified for human health hazards for Mutagenicity-Genetic Toxicity

Link to relevant study records

Reference

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

GLP compliance:

not specified

Type of assay:

other: Mouse micronucleus cytogenetic assay

Specific details on test material used for the study:

CAS Number: 28348-53-0
Identity: Cumene sulfonic acid, sodium salt
Purity: 99.4%
Remarks: 40% solution in water

Species:

mouse

Strain:

NMRI

Details on species / strain selection:

Species: Mouse , 24-30 g.
Strain: NMRI
Sex: Male/female
No. of animals 5 per sex per dose group per sample time

Sex:

male/female

Details on test animals or test system and environmental conditions:

Strain NMRI. Animals were approximately 24-30 g and acclimated for 1 week to the test conditions (20 =/- 3 degrees C, 30-70% relative humidity, 12 hour light/dark cycle).

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: water
- Justification for choice of solvent/vehicle: no data
- Concentration of test material in vehicle: 40 %
- Amount of vehicle: 16.8 ml/kg bw

Details on exposure:

Single oral dose at 0 and 4467 mg/kg bw; vehicle water, dose volume 17 ml/kg bw. Dose selection was based on preliminary studies with 2-5/sex: no
deaths at and below 4467 mg/kg bw ( one incidental death (female) at 3981 mg/kg bw), 4/10 deaths at 5000 mg/kg bw.

Duration of treatment / exposure:

72 hours

Frequency of treatment:

single oral application

Post exposure period:

24, 48 and 72 hours

Remarks:

Doses / Concentrations:
4467 mg/kg bw
Basis: actual ingested

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Positive control(s):

cyclophosphamide
- Justification for choice of positive control(s): no data
- Route of administration: oral by gavage
- Dose: 100 mg/kg bw
- Vehicle: water
- Total application volume: 10 ml/kg bw
- post exposure period: 24 hours

Tissues and cell types examined:

bone marrow; polychromatic erythrocytes (PCE), normochromatic erythrocytes (NCE)

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: The maximum tolerable dose (MTD)* was selected as dose.

The MTD was determined in a dose range finding study :
Phase 1: Limit test with 5000 mg/kg bw
Phase 2: Determination of the TMTD range with reduced animal number
Phase 3: Determination of the MTD with 5 animals/sex/dose

*MTD is defined as dose with no mortality but clear clinical symptoms within 3 days after application

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):
24, 48 and 72 hours after treatment the animals were killed by cervical dislocation and tissue was sampled.

DETAILS OF SLIDE PREPARATION:
The femora were removed and the bone marrow was suspended in fetal calf serum. The cell suspensions were centrifuged with 160 x g for 5 minutes and the supernatand discarded. The serum was resuspended and the suspension purified using a cellulose chromatographic column. The eluate was centrifuged at 800 x g for 10 minutes and the pellet in fetal calf serum /25 mM EDTA suspended. From this suspension 3-4 smears per animals were prepared on slides which were dried for at least 24 hours and stained with May-Grünwald/Giemsa solution.

METHOD OF ANALYSIS:
The cell analysis was performed by means of a Zeiss miscroscope at a 1000fold magnification (oil immersion). At least 1000 polychromatic erythrocytes (PCE) per animal were examined to determine the frequency of micronucleated cells. The ratio of PCE to normochromatic cells (NCO) was determined for a sample of 1000 erythrocytes. The number of micronucleated cells in counted NCE was determined.

Evaluation criteria:

For the identification of micronuclei the following criteria were considered:
a) roundish and clear contour by the nuclear membrane
b) diameter of about 1/20 of the size of the polychromatic erythrocyte
c) lays in the same focus layer as the observed erythrocyte

The micronucleus test is regarded as positive (test substance induces micronuclei in polychromatic erythrocytes) if the frequency of mucronucleated polychromatic erythrocytes of at least one tretament group is statistically significantly increased compared to the negative control and the increase is biologically relevant.

Statistics:

Mean values and standard deviations were calculated for the following parameters:
a) number of polychromatic erythrocytes (PCE) containing micronuclei
b) ratio of PCE/NCE

Comparison of treatment groups with different post exposure periods with negative controls of respective post exposure periods. After control of the relative frequency of micronuclei in the treatment groups on homogeneity with the mean relative frequency a statistical analysis of micronucleus frequency using a 2 x 2 contingency table with chi² test and continuity table according to Yates was performed (see [1]).
The differences of miconucleus frequencies in the positive control were reassessed in the two-sided t-test. This test was also used for the statistical analysis of the PCE/NCE-ratio.

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

Sodium cumenesulphonate under these test conditions is regarded as not mutagenic in the micronucleus test.

Conclusions:

Interpretation of results : negative
All male mice treated with the test substance showed no statistically significant increase in micronucleus frequency at any sampling time compared to control animals. For the female mice treated with the test substance at sampling times 24 and 48 hours after treatment also no statistically significant increase in micronucleus frequency was observed. Only at sampling time point 72 hours a statistically significant increase of polychromatic erythrocytes with micronuclei compared to control animals was observed. This effect was regarded as biologically not relevant as this increase ís based on the exceptional low micronucleus frequency of vehicle control group.
Sodium cumenesulphonate under these test conditions is regarded as not mutagenic in the micronucleus test.

Executive summary:

Interpretation of results : negative

All male mice treated with the test substance showed no statistically significant increase in micronucleus frequency at any sampling time compared to control animals. For the female mice treated with the test substance at sampling times 24 and 48 hours after treatment also no statistically significant increase in micronucleus frequency was observed. Only at sampling time point 72 hours a statistically significant increase of polychromatic erythrocytes with micronuclei compared to control animals was observed. This effect was regarded as biologically not relevant as this increase ís based on the exceptional low micronucleus frequency of vehicle control group.

Sodium cumenesulphonate under these test conditions is regarded as not mutagenic in the micronucleus test.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:  

 

The mutagenic potential of sodium cumene sulfonate, at 40% active ingredient was evaluated in the bacterial reverse mutation (Ames) assay using Salmonella typhimurium strains TA 98, 100, 1535, 1537 and 1538. There was no evidence of mutagenicity observed with and without metabolic activation.

Technical grade (65% a.i.) sodium xylene sulfonate was tested at 500 – 5000 μg/mL using Chinese hamster ovary cells (51). There were two independent tests with an exposure period of 25.5 hours.

The results indicated clastogenic activity (cell cycle delay) without metabolic activation at 2513 – 5000 μg/mL which was addressed by lengthening incubation time to 32.5 hours to ensure a sufficient number of scorable (second-division metaphase) cells. No clastogenic activity was recorded with metabolic activation.

No mutagenic activity in the (Q)SAR study, In vitro mutagenicity (Ames test) alerts by ISS for  sodium cumenesulphonate and does not cause in vitro mutagenicity (Ames test) .This QSAR method is Relevant for in vitro (Ames test) mutagenicity endpoints.

  

 Justification for selection of genetic toxicity endpoint Negative in all test conducted.

Additional information from genetic toxicity in vivo:  

Male mice treated with the test substance showed no statistically significant increase in micronucleus frequency at any sampling time compared to control animals. For the female mice treated with the test substance at sampling times 24 and 48 hours after treatment also no statistically significant increase in micronucleus frequency was observed. Only at sampling time point 72 hours a statistically significant increase of polychromatic erythrocytes with micronuclei compared to control animals was observed. This effect was regarded as biologically not relevant as this increase ís based on the exceptional low micronucleus frequency of vehicle control group.

Sodium cumenesulphonate under these test conditions is regarded as not mutagenic in the micronucleus test.

Justification for classification or non-classification

Based on the hazard assessment of Sodium cumenesulphonate in section 2.1 and 2.2. in IUCLID 6, available data for the substance and following the “Guidance on Information Requirement and Chemical Safety Assessment R.8. Characterisation of dose [concentration]- response for human health” andaccording to the criteria described in Directive 67/548 and in the CLP Regulation:

 

Directive 67/548	Mutagenicity-Genetic Toxicity Muta. Cat. 1; R46 May cause heritable genetic damage. Muta. Cat. 2; R46 May cause heritable genetic damage. Muta. Cat. 3; R68 Possible risk of irreversible effects.
CLP	Germ cell mutagenicity Muta. 1A Muta. 1B Muta. 2 H340: May cause genetic defects <state route of exposure if it is conclusively proven that no other routes of exposure cause the hazard>. H341: Suspected of causing genetic defects <state route of exposure if it is conclusively proven that no other routes of exposure cause the hazard>.

 

 

It is concluded that the substance Sodium cumenesulphonate does not meet the criteria to be classified for human health hazards for Mutagenicity-Genetic Toxicity