Reaction products of decanoic acid and lauric acid with glycerol and polyglycerol

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

consistently negative in the bacterial reverse mutation assay, mammalian cell gene mutation assay and mammalian cell chromosoem aberration assay

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1996-09 to 1996-10

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

26 th May, 1983

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Target gene:

Histidine locus

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

The test concentrations were: 0, 8, 40, 200, 1000 and 5000 µg/plate based on the results from the dose range finding test.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solubility of the test substance and low amount of spontaneous revertants within the control.

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

cumene hydroperoxide

other: 2-Aminoanthracene (with S9)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)
- Cell density at seeding (if applicable):1E09 cells/plate

DURATION
- Exposure duration: 65 h

NUMBER OF REPLICATIONS: 3

DETERMINATION OF CYTOTOXICITY
- Method: background bacterial lawn

Evaluation criteria:

A positive result is indicated by a dose response with statistically significant increases in revertant numbers

Statistics:

Mean standard deviations and Dunnett´s t-statistic

Key result

Species / strain:

S. typhimurium, other: TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

cytotoxic at 5000 µg/plate without S9, not cytotoxic with S9 up to limit concentration

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Table 1:Number of revertants per plate (mean of 3 plates)

	[strain 1535]		[strain 1537]		[strain 102]		[strain 98]		[strain 100]
Conc. [µg/plate]	— MA	+ MA	— MA	+ MA	— MA	+ MA	— MA	+ MA	— MA	+ MA
0*	8.3	11.7	8.0	6.0	228.7	209.0	19.3	44.0	81.3	90.0
8	15.3	11.7	9.3	7.5	229.7	259.0	22.0	40.0	91.0	91.3
40	12.7	8.7	10.0	8.0	215.7	253.7	21.0	34.7	100.7	84.0
200	18.7	9.7	7.0	11.5	192.7	214.3	27.3	31.3	93.7	91.7
1000	10.3	14.5	7.0	12.3	163.7	161.0	19.3	31.5	89.0	90.0
5000	b	5.0	b	3.0	a	89.3	b	20.3	a	64.7
Positive control	325.0	70.3	173.3	27.0	307.3	418.0	138.0	221.0	415.7	362.7

*solvent control with DMSO

b = Complete loss of lawn

a = Reduced lawn.

Conclusions:

Under the conditions of this Ames test, polyglycerin caprinate was negative for mutagenic potential.

Executive summary:

In a reverse gene mutation assay in bacteria according to OECD guideline 471 (adopted 21 July 1997), Salmonella typhimurium strains TA 1535, TA 1537, TA 102, TA 98 and TA 100 were exposed to polyglycerin caprinate in DMSO in concentrations of 0 (control), 8, 40, 200, 1000 and 5000 µg/plate in all strains in the presence and absence of mammalian metabolic activation (rat liver S9 mix). The assay was performed using the plate incorporation method.

The test substance was tested up to limit concentrations. Cytotoxic effects (reduced background lawn) were noted at 5000 µg/plate without S9; no cytotoxicity was observed with S9 up to the limit concentration. The positive controls induced the appropriate responses in the corresponding strains.

There was no evidence of a statistically significant increase in the number of revertant colonies in any of the five tester strains. Therefore, test substance was considered to be non-genotoxic (non-mutagenic) in Salmonella tester strains TA 1535, TA 1537, TA 102, TA 98 and TA 100 under the conditions employed (plate incorporation assay).

There was no evidence of induced mutant colonies over background. Under the conditions of the study, the test substance was negative for mutagenic potential.

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that source and target substances have similar physicochemical, ecotoxicological and toxicological properties because
• they are manufactured from similar or identical precursors under similar conditions
• they share structural similarities with common functional groups: the target and source substances are esterification products of fatty acids of varying chain lengths with glycerol and/or polyglycerol.
• the metabolism pathway leads to comparable products (glycerol and/or polyglycerol and medium or long chain fatty acids).

Therefore, read-across from the existing toxicity studies on the source substances is considered as an appropriate adaptation to the standard information requirements of REACH regulation

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
see “Justification for read-across” attached to IUCLID section 13

3. ANALOGUE APPROACH JUSTIFICATION
see “Justification for read-across” attached to IUCLID section 13

4. DATA MATRIX
see “Justification for read-across” attached to IUCLID section 13

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across: supporting information

Type of assay:

in vitro mammalian chromosome aberration test

Species / strain:

other: CHO; human lymphocytes

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

Conclusions:

Interpretation of results: negative with and without metabolic activation

Reference 3

Endpoint:

in vitro gene mutation study in mammalian cells

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that source and target substances have similar physicochemical, ecotoxicological and toxicological properties because
• they are manufactured from similar or identical precursors under similar conditions
• they share structural similarities with common functional groups: the target and source substances are esterification products of fatty acids of varying chain lengths with glycerol and/or polyglycerol.
• the metabolism pathway leads to comparable products (glycerol and/or polyglycerol and medium or long chain fatty acids).

Therefore, read-across from the existing toxicity studies on the source substances is considered as an appropriate adaptation to the standard information requirements of REACH regulation

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
see “Justification for read-across” attached to IUCLID section 13

3. ANALOGUE APPROACH JUSTIFICATION
see “Justification for read-across” attached to IUCLID section 13

4. DATA MATRIX
see “Justification for read-across” attached to IUCLID section 13

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across: supporting information

Type of assay:

mammalian cell gene mutation assay

Target gene:

HPRT

Species / strain:

Chinese hamster Ovary (CHO)

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

Conclusions:

Interpretation of results: negative with and without metabolic activation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

A bacterial reverse mutation assay was conducted with the target substance polyglycerin caprylate/caprinate. Additionally, genetic toxicity studies are available for the closely related source substances Medium- and long-chain triacylglycerols, short- and long-chain fatty acid triacylglycerols and1,2,3-Propanetriol, homopolymer, dodecanoate. A justification for read-across is given in iuclid section 13.

 

Bacterial reverse mutation assays

In a reverse gene mutation assay in bacteria according to OECD guideline 471 (adopted 21 July 1997), Salmonella typhimurium strains TA 1535, TA 1537, TA 102, TA 98 and TA 100 were exposed to polyglycerin caprinate in DMSO in concentrations of 0 (control), 8, 40, 200, 1000 and 5000 µg/plate in all strains in the presence and absence of mammalian metabolic activation (rat liver S9 mix). The assay was performed using the plate incorporation method.

The test substance was tested up to limit concentrations. Cytotoxic effects (reduced background lawn) were noted at 5000 µg/plate without S9; no cytotoxicity was observed with S9 up to the limit concentration. The positive controls induced the appropriate responses in the corresponding strains.

There was no evidence of a statistically significant increase in the number of revertant colonies in any of the five tester strains. Therefore, test substance was considered to be non-genotoxic (non-mutagenic) in Salmonella tester strains TA 1535, TA 1537, TA 102, TA 98 and TA 100 under the conditions employed (plate incorporation assay).

There was no evidence of induced mutant colonies over background. Under the conditions of the study, the test substance was negative for mutagenic potential.

 

Medium- and long-chain triacylglycerols (MLCT) did not induce gene mutations in 4 different Salmonella (S. typhimurium TA 1535, TA 1537, TA 98 and TA 100) and one E. coli strain (E. coli WP2 uvr A) when tested up to limit concentrations (0, 313, 625, 1250, 2500, 5000 µg/plate).

 

in vitro gene mutation study in mammalian cells

An HPRT test was conducted withshort- and long-chain fatty acid triacylglycerols (SALATRIM) in Chinese hamster Ovary (CHO) cells at concentrations of 0, 31.25, 62.5, 125, 250, 500 and 1000 μg/mL. The high dose was limited by the low solubility of the fats in the assay medium. There was no evidence of mutagenic potential under the conditions of the assay.

 

Chromosome aberration assays

An in vitro mammalian chromosome aberration test was conducted withshort- and long-chain fatty acid triacylglycerols (SALATRIM) Chinese hamster Ovary (CHO) cells at concentrations of 250, 500 and 1000 µg/mL ion acetone.

There was no change in the mitotic index with either fat compared to the solvent control. Neither fat produced increases in the percent cells with abnormal chromosomes or in any of the other criteria for chromosomal damage. Therefore, short- and long-chain fatty acid triacylglycerols were neither clastogenic nor cytotoxic in the chromosomal aberration assay in the presence or absence of metabolic activation.

 

An in vitro mammalian chromosome aberration test was conducted with 1,2,3-Propanetriol, homopolymer, dodecanoate in Chinese hamster lung fibroblasts (V79) up to cytotoxic concentrations.

Without metabolic activation a slight increase of aberrant cells was observed at concentrations of 50 and 70 µg/mL but not at 65 µg/mL. With metabolic activation, the aberration rates of the higher concentrations (1250 and 1500 µg/mL) were significantly increased and a dose relationship was observed.

The test substance was clastogenic to chinese hamster V79 cells treated in vitro under the conditions of the test.

However, cytotoxicity was observed at concentrations ≥ 65 µg/mL without metabolic activation and at concentrations ≥ 1250 µg/mL with metabolic activation. Thus, chromosome aberrations were only observed in the presence of cytotoxic effects.

 

A second in vitro mammalian chromosome aberration test was conducted with 1,2,3-Propanetriol, homopolymer, dodecanoate in human lymphocytes up to cytotoxic concentrations

No significant increases in chromosomal aberrant aberrations were observed in any treatment group at any dose level. The test substance was not clastogenic to human lymphocytes treated in vitro under the conditions of the test.

Considering that the second study was negative and the first positive only in the presence of marked cytotoxicity, the first chromosome aberration test can be considered a false positive.

 

Conclusion

Overall, the target substancePolyglycerin caprylate/caprinate is neither mutagenic nor clastogenic in vitro based on the Ames test conducted with the target substance and the mammalian cell gene mutation assay as well as the chromosome aberration tests conducted with closely related source substances.

 

There are no data gaps for the endpoint genetic toxicity. There is no reason to believe that the results would not be relevant to humans. 

Justification for classification or non-classification

Based on the available data, polyglycerin caprylate/caprinate does not need to be classified for mutagenicity according to regulation (EC) 1272/2008. Thus, no labelling is required.