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Key value for chemical safety assessment

Additional information

All key studies for this endpoint addressing bacterial mutagenicity, mammalian mutagenicity and mammalian clastogenicity were all negative for either o/p-TSA itself or via cross-reading from both its individual constituents p-TSA and o-TSA. Furthermore, substance has no structural alerts for mutagenicity, and a large database of performed studies indicate that neither p-TSA nor o-TSA possess genotoxic properties.

 

Discussion:

o/p-TSA consists of a mixture of p-TSA and o-TSA. Both these substances show no structural alerts for mutagenicity, and a large database of performed studies indicate that p-TSA and o-TSA do not have genotoxic properties.

 

The table below lists all available data, including from literature:


Type of test

Substance

Test system

Doses

Results

Reference 

Bacterial test

Ames test (reverse mutation)

o/p-TSA

S. typhimuriumstrains TA98, TA100, TA1535, TA1537, TA1538;Saccharomyces cerevisiaeD4

1.0 µg to 1000 µg/plate

No mutagenic effects observed (+/- S9).

Proprietary: Litton Bionetics, 1978 (Ketjenflex 9R)

Ames test (reverse mutation)

o/p-TSA

S. typhimuriumstrains TA98, TA100, TA1535, TA1537, TA1538;Saccharomyces cerevisiaeD4

0.5 µ to 250 µg/plate

No mutagenic effects observed (+/- S9).

Proprietary: Litton Bionetics, 1976

Ames test (reverse mutation)

p-TSA

S. typhimuriumstrains TA98, TA100, TA1535, TA1537, TA1538;Saccharomyces cerevisiaeD4

1, 10, 100, 500, 1000 µg/plate

No mutagenic effects observed (+/- S9).

Proprietary: Litton Bionetics, 1978 (o-TSA)

Ames test (reverse mutation)

p-TSA

S. typhimuriumstrains TA98, TA100, TA1535, TA1537;Escherichia coliWP2 uvrA

0, 312.5, 625, 1250, 2500, 5000 μg/plate

No mutagenic effects (+/- S9).

MHW, Japan; cited by OECD (1994)

Ames test (reverse mutation)

p-TSA

S. typhimuriumstrains TA98, TA100, TA102

TA98 &100: 0, 33, 100, 333, 1000 and 2000 or 3333 µg/plate ; TA102: 0, 100, 333, 1000, 3333 and 10000 µg/plate

No mutagenic effects observed (+/- S9).

NTP, 2007

Ames test (reverse mutation)

p-TSA

S. typhimuriumstrains TA1530, TA1535, TA1538, TA98, and TA100

4 x 10-7 mol up to 4 x 10-2 mol/plate.

No direct mutagenic effects were observed (only +S9).

Poncelet et al. (1980)

Ames test (reverse mutation)

p-TSA

S. typhimuriumstrainTA98, TA100, TA1535, TA1537

Up to 18000 μg/plate

No mutagenic activity (+/- S9). Evaluations included testing in ZLM agar (similar condition Eckhardt, 1981)

Herbold (1981)

Ames test (reverse mutation)

p-TSA

S. typhimurium –TA98, TA100, TA1535, TA1537 and TA1538.

Doses of up to 12 mg/plate were used.

(+/- S9) The only positive (marginally) increase in revertants were found intester strain TA98, with S9 and only using ZLM agar, and not in VB agar.

Eckhardt et al. (1980)

Gene mutation assay

o-TSA

S. typhimuriumstrains TA98, TA100, TA1535, TA1537, TA1538;Saccharomyces cerevisiaeD4

Up to 1000 ug/plate

Negative (+/- S9)

Proprietary, Litton Bionetics, 20838, 1978

Ames test (reverse mutation)

o-TSA

S. typhimurium (TA98, TA100, TA1535, TA1537) E. coli (WP2uvr A)

Up to 5000 ug/plate

Negative (+/- S9)

MHW, Japan: 1999

Ames test (reverse mutation)

o-TSA

S. typhimurium (TA1535, TA100, TA98, TA1537)

Up to 1000 ug/plate

Negative (+/- S9)

Stolts et al.: 1977

Ames test (reverse mutation)

o-TSA

S. typhimurium (TA1530, TA1535, TA1537, TA1538, TA100, TA98)

Up to 4 x 10-2 mol/plate

Negative (+/- S9)

Poncelet et al.: 1979

Ames test (reverse mutation)

o-TSA

S. typhimurium (TA1535, TA100, TA1538, TA98, TA1537)

Up to 18000 ug/plate

Weakly positive: TA98 (+ S9)
Negative: TA98 (-S9) and other strains (+/- S9)

Eckhardt et al.: 1980

Ames test (reverse mutation)

o-TSA

S. typhimurium (TA98, TA 100, TA 1535, TA1537)

Up to 18000 ug/plate

Negative (+S9)

Herbold: 1981

Ames test (reverse mutation)

o-TSA

S. typhimurium (TA1535, TA1538, TA98, TA100)

Up to 2500 ug/plate

Negative (+S9)

Ashby et al.: 1978

Non-bacterial in vitro test

forward mutations (TK locus)

p-TSA

L5 178Y mouse lymphoma cells

Up to 5000 µg/mL

(+/- S9) An increased frequency of mutations was only observed at the highest, cytotoxic, dose level with metabolic activation, involving an increase in small colonies.

Proprietary. Covance, 2000

Chromosomal aberration

p-TSA

CHL cells

Without S9: 0, 0.33, 0.65, 1.30 mg/mL; with S9: 0, 0.43, 0.85, 1.70 mg/mL.

No chromosomal aberrations (+/- S9).

MHW, Japan; cited by OECD (1994)

Chromosomal aberration

p-TSA

CHO-K1 cells

Cells were treated with 14, 200, and 400 μg/mL p-TSA for 24 h.

(- S9) Negative.

Masubuchi (1978a abstr)

Development of ouabain resistance

p-TSA

RSa human cell line

900 and 1800 μg/mL [5.26 and 10.51 mM], for 24 h

(- S9) No increase in ouabain resistance was observed inp-TSAtreated cells, relative to controls.

Suzuki and Suzuki (1988)

forward mutations (TK locus)

o-TSA

mouse lymphoma L5178Y cells

Up to 1712 μg/mL

Negative (+/- S9)

Proprietary, TNO Triskelion, V20203/05, 2013

Chromosomal aberration test

o-TSA

CHL cells

Up to 3000 ug/mL

Negative (+/- S9)

MHW, Japan:1999

Ouabain-resistant mutation assay

o-TSA

Human RSa cells

Up to 1800 ug/mL

Negative (-S9)

Suzuki & Suzuki: 1988

Chromosomal aberration test

o-TSA

CHO-K1 cells

Up to 400 ug/mL

Negative (-S9)

Masubuchi et al.: 1978

In vivotest

Micronucleus assay

p-TSA

Mouse Crl:CD-1, adult, M and F

Oral 187.5, 375, 750, 1500 mg/kg bw

Negative

Proprietary. Covance, 2002

Micronucleus assay

p-TSA

Mouse Crl:CD-1, adult, only males

i.v. 80, 120, 160 mg/kg bw,

Negative

Proprietary. Covance, 2000

Micronucleus assay

p-TSA

Mouse, NMRI, adult, number n.p., M and F

p-TSA administered both orally 2 x 855 mg/kg and i.p. 2 x 428 and 2 x 855 mg/kg

Negative

Eckhardt et al. (1980)

Mammalian spot test

o-TSA

Mouse

1000 mg/kg bw (oral)

Inconclusive

Fahrig: 1982

Micronucleus assay

o-TSA

Mouse

1026 mg/kg bw (by gavage)

Negative

Eckhardt et al.: 1980

Micronucleus assay

o-TSA

Mouse

Up to 1026 mg/kg bw (i.p.)

Negative

Eckhardt et al.: 1980

SLRL (Sex-linked recessive lethal) test

p-TSA

D. melanogaster(Berlin K [wild-type] and Basc strains)

Conc. 2.5 mM in 5% sucrose solutions used as feed for 3 days.

A small but statistically significant increase in the frequency of recessive lethal mutations was observed only in the first brood, corresponding to mature sperm. (about double the control frequency)

Eckhardt et al. (1980)

SLRL (Sex- linked recessive lethal) test

p-TSA

D. melanogaster(Oregon-K [wild type] and Basc strains)

Adult flies injected abdominal with about 0.2 μL of a 5 mM solution (~0.170 μg)

There were no or marginal positive effects observed in first brood under the test conditions. (frequency was about 2.5 times higher than of the control)

Kramers (1977)

 

References:

o/p-TSA

·   Proprietary: Litton Bionetics (Jagannath, D. R. & Brusick, D.), 20838, 1978.(Ketjenflex 9R)

·   Proprietary: Litton Bionetics (Brusick, D.), 2547, 1976.

o-TSA

·   Proprietary: Litton Bionetics (Jagannath, D. R. & Brusick, D.), 20838, 1978.(o-TSA)

·   Proprietary: TNO Triskelion, V20203/05, 2013

·   Ashby, J. et al. (1978) Fd. Cosmet. Toxicol., 16, 95-103.

·   Eckhardt, K. et al. (1980) Toxicology Letters, 7, 51-60.

·   Fahrig, R. (1982) Mutat. Res., 103, 43-47.

·   Herbold, B.A. (1981) Mutat. Res., 90, 365-372.

·   Masubuchi, M. et al. (1978) Mutat. Res., 54, 242-243.

·   MHW, Japan (1999) Ministry of Health and Welfare, Toxicity Testing Reports of Environmental Chemicals, 7, 125-160.

·   Poncelet, F. et al. (1979) Fd. Cosmet. Toxcol., 17, 229-231.

·   Stolts, D. R. et al. (1977) J. Environ. Pathol. Toxicol., 1, 139-146

·   Suzuki, H. & Suzuki, N. (1988) Mutat. Res., 209, 13-16.

p-TSA:

·   Covance, 2000, Proprietary: in vitro-Mouse Lymphoma, report# 21198-0-431-ICH

·   Covance, 2000, Proprietary: in vivo-micronucleus, report# 21198-0-455OECD

·   Covance, 2002, Proprietary: in vivo-micronucleus, report# 3464-0-455OECD

·   Eckhardt K., M.T. King, E. Gocke, and D. Wild. 1980. Mutagenicity study of Remsen-Fahlberg saccharin and contaminants. Toxicol Lett 7:51-60

·   Herbold B.A. 1981 . Studies to evaluate artificial sweeteners, especially Remsen-Fahlberg saccharin, and their possible impurities, for potential mutagenicity by the Salmonella mammalian liver microsome test. Mutat Res 90:365-372.

·   Kramers P.G. 1977. Mutagenicity of saccharin in Drosophila: The possible role of contaminants. Mutat Res 56:163-167

·   Litton Bionetics, 1978, Proprietary: in vitro-Ames, report# 20838

·   Masubuchi, M., S. Nawai, M. Hirokado, and K. Hiraga. 1978a abstr. Lack of the cytogenetic effects of saccharin impurities on CHO-K1 cells. Mutat. Res. 54[2], 242-243.

·   MHW, Japan; cited by OECD (1994) – Ames: Hatano Research Institute, Food and Drug Safety Center, Japan, Reverse Mutation Test of 4 -Methylbenzenesulfonamide on Bacteria (http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF70-55-3e.pdf)

·   MHW, Japan; cited by OECD (1994) – CHL: Hatano Research Institute, Food and Drug Safety Center, Japan, In vitro Chromosomal Aberration Test of 4-Methylbenzenesulfonamide on Cultured Chinese Hamster Cells (http://dra4.nihs.go.jp/mhlw_data/home/pdf/PDF70-55-3f.pdf)

·   NTP, 2007: The Salmonella Mutagenicity Test on p-Toluenesulfonamide, report A84238 (http://tools.niehs.nih.gov/ntp_tox)

·   Poncelet F., M. Mercier, and J. Lederer. 1980. Saccharin: Para forms of some impurities are not mutagenic inSalmonella typhimurium. Food Cosmet

·   Suzuki H., and N. Suzuki. 1988. Mutagenicity of saccharin in a human cell strain. Mutat Res 209:13-16.


Justification for selection of genetic toxicity endpoint
No one specific study is selected. All key studies for this endpoint addressing bacterial mutagenicity, mammalian mutagenicity and mammalian clastogenicity were all negative.

Short description of key information:
All key studies for this endpoint addressing bacterial mutagenicity, mammalian mutagenicity and mammalian clastogenicity were all negative for either o/p-TSA itself or via cross-reading from both its individual consituents p-TSA and o-TSA. Furthermore, substance has no structural alerts for mutagenicity, and a large database of performed studies indicate that neither p-TSA nor o-TSA possess genotoxic properties.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

Both p-TSA and o-TSA have no structural alerts for mutagenicity, and available studies covering bacterial mutagenicity, mammalian mutagenicity and mammalian clastogenicity all indicate that p-TSA and o-TSA do not have genotoxic properties. Consequently their mixture o/p-TSA does not need to be classified either.