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Endpoint summary

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Administrative data

Key value for chemical safety assessment

Additional information

Justification for grouping of substances and read-across

The Glycerides category covers aliphatic (fatty) acid esters of glycerol. The category contains both well-defined and UVCB substances with aliphatic acid carbon chain lengths of C2 (acetate) and C7-C22, which are mostly linear saturated and even numbered. Some of the substances in the category contain unsaturated fatty acids (e.g. oleic acid in 2,3-dihydroxypropyl oleate, CAS 111-03-5 or general fatty acids C16-22 (even) unsaturated in Glycerides, C14-18 and C16-22-unsatd., mono- and di-, CAS 91744-43-7). Some category members contain branched fatty acids. Branching is mostly methyl groups (e.g. isooctadecanoic acid, monoester with glycerol, CAS 66085-00-5 or 1,2,3-propanetriyl triisooctadecanoate, CAS 26942-95-0). In one category member the branching cannot be precisely located (Glycerides, C16-18 and C18-unsatd., branched and linear mono-, di- and tri, ELINCS 460-300-6). Hydroxylated fatty acids are present in three substances (Castor oil, CAS 8001-79-4; castor oil hydrogenated, CAS 8001-78-3 and 2,3-dihydroxypropyl 12-hydroxyoctadecanoate, CAS 6284-43-1). Hydroxylation occurs on C12 of stearic acid in all these substances. Acetylated chains are present in the last part of the category, comprising fatty acids from C8 to C18 (even) and also C18 unsaturated, additionally a C18 acetylated fatty acid is present with the acetic acid located in C12 position (e.g. Glycerides, castor oil mono-, hydrogenated acetates / 12-acetoxy-octadecanoic acid, 2,3-diacetoxy, CAS 736150-63-3). All glycerides build mono-, di- and tri-esters in variable proportions.

The available data allows for an accurate hazard and risk assessment of the category and the category concept is applied for the assessment of environmental fate, environmental and human health hazards. Thus where applicable, environmental and human health effects are predicted from adequate and reliable data for source substance(s) within the group by interpolation to the target substances in the group (read-across approach) applying the group concept in accordance with Annex XI, Item 1.5, of Regulation (EC) No 1907/2006. In particular, for each specific endpoint the source substance(s) structurally closest to the target substance is/are chosen for read-across, with due regard to the requirements of adequacy and reliability of the available data. Structural similarities and similarities in properties and/or activities of the source and target substance are the basis of read-across.

A detailed justification for the grouping of chemicals and read-across is provided in the technical dossier (see IUCLID Section 13) and within Chapter 5.1 of the CSR.

 

Overview of Genetic toxicity

CAS

Genetic Toxicity in vitro: gene mutation in bacteria

Genetic Toxicity in vitro: cytogenicity in mammalian cells

Genetic Toxicity in vitro: gene mutation in mammalian cells

Genetic toxicity in vivo

26402-26-6 (b)

Experimental result:
not mutagenic

--

--

--

142-18-7 (a)

WoE:
Experimental result:
not mutagenic
RA: CAS 91845-19-1
RA: CAS 97593-30-1 (C12)

WoE:
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-85-1
RA: CAS 111-14-8

WoE:
RA: CAS 736150-63-3
RA: CAS 56-85-1
RA: CAS 111-14-8

--

111-03-5

Experimental result:
not mutagenic

Experimental result:
not clastogenic

--

--

6284-43-1

WoE:
RA: CAS 91845-19-1
RA: CAS 736150-63-3

WoE:
RA: CAS 736150-63-3
RA: CAS 8001-79-4

RA: CAS 736150-63-3

--

620-67-7

WoE:
RA: CAS 538-23-8
RA: CAS 56-85-1
RA: CAS 111-14-8

WoE:
RA: CAS 91052-13-0
RA: CAS 56-85-1
RA: CAS 111-14-8

WoE
RA: CAS 56-85-1
RA: CAS 111-14-8

--

538-23-8

Experimental result:
not mutagenic

--

--

WoE:
Experimental result:
not clastogenic

122-32-7

WoE:
Experimental result:
not mutagenic
RA: CAS 111-03-5
RA: MLCT

WoE:
RA: CAS 111-03-5
RA: CAS 8001-79-4

WoE:
RA: CAS 736150-63-3
RA: CAS 56-85-1
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 91845-19-1
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

555-43-1

WoE:
Experimental result:
not mutagenic
RA: CAS 736150-63-3
RA: ELINCS 460-300-6

WoE:
RA: CAS 736150-63-3
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE
RA: CAS 736150-63-3

--

26942-95-0

Experimental result:
not mutagenic

--

--

--

91052-47-0

WoE:
RA: CAS 91744-13-7
RA: MLCT
RA: CAS 736150-63-3
RA: ELINCS 460-300-6

WoE:
RA: CAS 8001-79-4
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 91845-19-1
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

91744-09-1

WoE:
RA: CAS 91744-13-7
RA: MLCT
RA: CAS 91052-13-0
RA: ELINCS 460-300-6
RA: CAS 111-03-5

WoE:
RA: CAS 8001-79-4
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 91845-19-1
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

85536-07-8

WoE:
RA: CAS 97593-30-1 (C12)
RA: CAS 538-23-8
RA: CAS 73398-61-5
RA: CAS 26402-26-6
RA: CAS 65381-09-1

WoE:
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8

--

91052-49-2

WoE:
RA: CAS 91052-13-0
RA: CAS 97593-30-1 (C12)
RA: CAS 736150-63-3
RA: CAS 67701-26-2

WoE:
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8

--

67701-33-1

WoE:
RA: CAS 91744-13-7
RA: CAS 91052-13-0
RA: ELINCS 460-300-6
RA: MLCT

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 8001-79-4
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 91845-19-1
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

67784-87-6

WoE:
RA: CAS 736150-63-3
RA: ELINCS 460-300-6

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 91845-19-1
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

91845-19-1

Experimental result:
not mutagenic

--

--

Experimental result:
not clastogenic

97358-80-0

WoE:
RA: CAS 736150-63-3
RA: ELINCS 460-300-6
RA: CAS 26942-95-0

WoE:
RA: CAS 736150-63-3
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

--

91744-13-7

Experimental result:
not mutagenic

--

--

--

31566-31-1

WoE:
RA: CAS 91744-13-7
RA: MLCT
RA: CAS 736150-63-3
RA: ELINCS 460-300-6

WoE:
RA: CAS 8001-79-4
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 91845-19-1
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

85251-77-0

WoE:
RA: CAS 91744-13-7
RA: MLCT
RA: CAS 736150-63-3
RA: ELINCS 460-300-6

WoE:
RA: CAS 8001-79-4
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 91845-19-1
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

91052-28-7

WoE:
RA: CAS 91744-13-7
RA: CAS 91052-13-0
RA: CAS 111-03-5
RA: ELINCS 460-300-6
RA: MLCT

WoE:
RA: CAS 736150-63-3
RA: CAS 8001-79-4
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: CAS 111-03-5
RA: CAS 112-85-6
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 91845-19-1
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

91052-54-9

WoE:
Experimental result:
not mutagenic
RA: CAS 736150-63-3
RA: ELINCS 460-300-6

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 91845-19-1
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

91744-20-6

WoE:
RA: CAS 91744-13-7
RA: MLCT
RA: CAS 91052-13-0
RA: ELINCS 460-300-6
RA: CAS 111-03-5

WoE:
RA: CAS 8001-79-4
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 91845-19-1
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

no CAS

ELINCS 460-300-6: Glycerides, C16-C18 and C18-unsaturated, branched and linear mono-, di- and tri-

Experimental result:
not mutagenic

--

--

--

97722-02-6

WoE:
RA: CAS 91744-13-7
RA: MLCT
RA: CAS 91052-13-0
RA: ELINCS 460-300-6
RA: CAS 111-03-5

WoE:
RA: CAS 8001-79-4
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 8001-79-4
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

77538-19-3

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5

--

91744-28-4

WoE:
RA: CAS 91052-13-0
RA: CAS 97593-30-1 (C12)
RA: CAS 736150-63-3
RA: CAS 67701-26-2

WoE:
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8

--

68606-18-8

WoE:
RA: CAS 91052-13-0
RA: CAS 97593-30-1 (C12)
RA: CAS 538-23-8
RA: CAS 73398-61-5
RA: CAS 26402-26-6
RA: CAS 65381-09-1

WoE:
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8

--

65381-09-1

Experimental result:
not mutagenic

--

--

--

73398-61-5

WoE:
Experimental result:
not mutagenic
RA: CAS 555-43-1

WoE:
RA: CAS 8001-79-4
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 8001-79-4
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

85536-06-7

WoE:
RA: CAS 91052-13-0
RA: CAS 97593-30-1 (C12)
RA: CAS 538-23-8
RA: CAS 73398-61-5
RA: CAS 65381-09-1
RA: CAS 67701-26-2
RA: CAS 91052-54-9
RA: CAS 555-43-1

WoE:
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

--

67701-26-2

WoE:
Experimental result:
not mutagenic
RA: CAS 555-43-1
RA: CAS 91052-13-0
RA: CAS 73398-61-5
RA: CAS 91052-54-9
RA: MLCT

WoE:
RA: CAS 8001-79-4
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 8001-79-4
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

8001-79-4

--

Experimental result:
not clastogenic

Experimental result:
not mutagenic

Experimental result:
not clastogenic

8001-78-3

WoE:
Experimental result:
not mutagenic
RA: MLCT

WoE:
RA: CAS 8001-79-4
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

WoE:
RA: CAS 8001-79-4
RA: short- and long-chain triacylglycerols (SALATRIM 234CA and CS)

97593-30-1 (C10 )

RA: CAS 97593-30-1 (C12)

WoE:
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8

--

97593-30-1 (C12)

Experimental result:
not mutagenic

WoE:
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 111-14-8

--

93572-32-8

WoE:
RA: CAS 91052-13-0
RA: CAS 97593-30-1 (C12)
RA: CAS 736150-63-3

WoE:
RA: CAS 91052-13-0
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: CAS 112-85-6
RA: CAS 111-14-8

WoE:
RA: CAS 736150-63-3
RA: CAS 56-81-5
RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)
RA: CAS 111-14-8

--

91052-13-0

Experimental result:
not mutagenic

Experimental result:
not clastogenic

RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 23CA)

RA: short- and long-chain fatty acid triacylglycerols (SALATRIM 234CA and 234CS)

736150-63-3

Experimental result:
not mutagenic

Experimental result:
not clastogenic

Experimental result:
not mutagenic

Experimental result:
not clastogenic

no CAS

Short-, medium- and long-chain triglycerides (SCT, MCT, LCT) (c, d)

Experimental result:
not mutagenic

Experimental result:
not clastogenic

Experimental result:
not mutagenic

Experimental result:
not clastogenic

56-81-5 (c)

Experimental result:
not mutagenic

Experimental result:
not clastogenic

Experimental result:
not mutagenic

--

111-14-8 (c)

Experimental result:
not mutagenic

Experimental result:
not clastogenic

Experimental result:
not mutagenic

--

112-85-6 (c)

Experimental result:
not mutagenic

Experimental result:
not clastogenic

--

--

(a) Category members subject to the REACh Phase-in registration deadline of 31 May 2013 are indicated in bold font.

(b) Substances that are either already registered under REACh, or not subject to the REACh Phase-in registration deadline of 31 May 2013, are indicated in normal font.

(c) Surrogate substances are either chemicals forming part of a related category of structurally similar fatty acid esters or precursors/breakdown products of category members (i.e. alcohol and fatty acid moieties). Available data on these substances are used for assessment of (eco )toxicological properties by read-across on the same basis of structural similarity and/or mechanistic reasoning as described below for the present category.

(d) Assessment of toxicological properties is conducted also taking into account available data on mixtures of synthetic and/or naturally occurring glycerides (e.g. vegetable oils), which cannot be identified by a (single) CAS/EC number. The test materials short-, medium- and long-chain triglycerides (SCT, MCT, LCT) and their combinations (e.g. MLCT, SALATRIM – a SLCT) comprise triesters of glycerol with fatty acid chain lengths of C2 and C4 (short-chain), C8 and C10 (medium-chain) and C18 saturated/unsaturated (long-chain). The substance “mixture of mono-, di-, and triglycerides of lauric acid” comprises mono-, di and triesters of glycerol with dodecanoic acid (C12). The substance “Modified triglyceride” contains main components: 1,3-dioleoyl 2-palmitoyl triacylglycerol and 1,2-dipalmitoyl 3-oleoyl triacylglycerol, comprising triesters of glycerol with hexadecanoic (C16) and (9Z)-Octadec-9-enoic acid (C18:1). Available data on identity and composition of the individual test material for a given study is provided in the technical dossier.

For all category members registered under REACh a full data set for each endpoint is provided. For substances not subject to the current REACh Phase-in registration, lack of data for a given endpoint is indicated by "--".

 

Genetic toxicity (mutagenicity) in bacteria in vitro

CAS No. 26402-26-6

Octanoic acid, monoester with glycerol was investigated in a bacterial mutation assay similar to OECD guideline 471 and in compliance with GLP (Marquardt, 1995). Based on a preliminary cytotoxicity test, concentrations from 10 to 500 µg/plate were chosen for treatment of S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100 in the main study. The test material was not cytotoxic in any tester strain up to 100 µg/plate with and without S9-mix. Cytotoxic effects were observed in tester strains TA 98, TA 100, TA 1535 and TA 1537 at ≥ 250 µg/plate without S9-mix. At 500 µg/plate and with S9-mix, the test material was cytotoxic in tester strains TA 98 and TA 100. No increase in the mean number of revertants was observed in in the main experiment independent of metabolic activation and test substance concentration. The positive and negative controls verified the validity of the assay. Under the conditions of the study, the test substance was non-mutagenic in the presence or absence of metabolic activation.

CAS No. 142-18-7

A bacterial gene mutation assay with 2,3-dihydroxypropyl laurate was conducted in compliance with OECD guideline 471 and GLP (Banduhn, 1995). Two independent experiments were conducted both in the absence and in the presence of liver microsomal activation system (S9 mix). In the first experiment, S. typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 were treated with concentrations ranging from 8 to 5000 µg/plate. In the second experiment, concentrations ranging from 6.25 to 100 µg/plate were tested. In both experiments, cytotoxicity was observed at concentrations ≥ 40 µg/plate in the bacterial strains. The mean number of revertants was not increased at any concentrations tested. The positive and negative controls included revealed the expected results. Under the experimental conditions reported, the test substance did not induce mutations in the selected strains of S. typhimurium with or without metabolic activation.

CAS No. 111-03-5

In a GLP-study performed according to OECD guideline 471, the 2,3-dihydroxypropyl oleate was investigated in S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 and in E. coli WP2 uvrA (Nakajima, 2005). In three dose finding tests with test concentrations up to 5000 µg/plate, suitable concentrations for treatment of the bacterial strains in the main study were selected. Based on the results obtained in the pre-test regarding solubility and/or cytotoxicity of the test substance, concentrations up to 9.77 (TA 1537), 156 (TA 98 and TA 100) and 5000 µg/plate (TA 1535, WP2 uvrA) were chosen in the absence of metabolic activation, whereas concentrations up to 313 (TA 1537), 625 (TA 100) and 5000 µg/plate (TA 1535, WP2 uvrA and TA 98) were chosen in the presence of metabolic activation. Cytotoxic effects were present at 4.88 (TA 1537), 78.1 (TA 98) and 156 (TA 100) µg/plate without metabolic activation as well as at 156 (TA 100) and 313 (TA 1537) µg/plate with metabolic activation. Precipitation was noted at the end of the exposure period at ≥ 625 and ≥ 1250 µg/plate in the presence and absence of metabolic activation, respectively. No increase in the mean number of revertants per plate was observed in any strain compared to controls. The positive and negative controls included in the assay demonstrated the validity of the assay. Under the conditions of this assay, the test substance was not mutagenic in the presence or absence of metabolic activation.

CAS No. 538-23-8

Glycerol trioctanoate was investigated in a bacterial mutation assay performed similarly to OECD guideline 471 (NTP, 1989). In this study, four S. typhimurium strains were exposed to concentrations from 100 to 10000 µg/plate (TA 97, TA 98 and TA 100) and 100 to 16666 µg/plate (TA 1535), respectively. The bacteria strains were treated with the test substance for in the presence and absence of a hamster or rat liver metabolic activation system, respectively. No signs of cytotoxicity were observed. The mean number of revertants was not increased in TA 97, TA 98 and TA 100 strains, but was significantly and dose-dependently increased in TA 1535 with S9-mix at concentrations ≥ 6666 µg/plate compared to controls. The positive and negative controls included revealed the expected results. Under the conditions of this test, the test substance was non-mutagenic in S. typhimurium TA 97, TA 98 and TA 100, but showed positive results in the strain TA 1535 at very high concentrations. Thus, the results of this assay are considered to be ambiguous.

CAS No. 122-32-7

9-Octadecenoic acid (Z)-, 1,2,3-propanetriyl ester was tested in an Ames test performed similarly to OECD 471 (Nestmann et al., 1980). The experiment was performed in S. typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 at a concentration of 1000 µg/plate, both in the absence and in the presence of a liver microsomal activation system (S9 mix). No increase in the number of revertants was observed at the test concentration tested. However, solubility properties of the test substance did not allow testing at concentrations that might induce cytotoxicity in the bacteria. The positive and negative controls included into the study revealed the expected results. Under the conditions of this experiment, the test substance did not induce mutations either with or without metabolic activation.

CAS No. 555-43-1

A bacterial gene mutation assay with Glycerol tristearate was performed similarly to OECD guideline 471 and under GLP conditions (Ebert, 1997). Two independent experiments were performed both in the absence and in the presence of liver microsomal activation system (S9-mix). In both experiments, S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 were exposed at a concentration of 5000 µg/plate. No cytotoxicity and no increase in the mean number of revertants were observed. The positive and negative controls included showed the expected results. Under the experimental conditions reported, the test substance did not induce mutations in the absence and presence of metabolic activation in the selected strains.

CAS No. 26942-95-0

The potential mutagenicity of 1,2,3-propanetriyl triisooctadecanoate was assessed in four S. typhimurium strains (TA 1535, TA 1537, TA 98 and TA 100) in an Ames test similar to OECD Guideline 471 and under GLP conditions (Waart van de, 1996). Concentrations ranging from 100 to 5000 µg/plate were selected for treatment in the main assay. Precipitation was observed in the first experiment at the maximum concentration of 5000 µg/plate, which however did not influence the counting of revertant colonies. No increase in the mean number of revertants per plate was observed when compared to controls. The positive and negative controls included for each tester strain showed the expected results. Based on the study results, the test substance was considered non-mutagenic in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

CAS No. 91845-19-1

A bacterial gene mutation assay with Glycerides, C16-18 and C18-hydroxy mono- and di- was performed in compliance with OECD guideline 471 and GLP (Wallat, 1984). In two independent experiments, S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100 were exposed to concentrations from 8 to 5000 µg/plate with and without metabolic activation. At the maximum concentration, bacterial growth was completely inhibited in Salmonella strain TA 1537 in the presence (first experiment) or absence (second experiment) of metabolic activation, respectively. No increase in the mean number of revertants per plate was observed in any tester strain at the indicated concentrations. The vehicle controls were within the spontaneous mutation ranges of the historical controls and the positive controls induced the expected increase in the number of reverse mutants. Under the conditions of this experiment, the test substance did not show mutagenicity in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

CAS No. 91744-13-7

In a GLP-study performed according to OECD guideline 471, Glycerides, C14-18 and C16-22-unsatd. mono- and di- was investigated in S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100 (Banduhn, 1990). The tester strains were exposed to concentrations ranging from 8 to 5000 µg/plate in the presence or absence of a metabolic activation system (S9-mix). After exposure, a partial inhibition of bacterial growth, as indicated by the reduction in bacterial background lawn, was observed in TA 1535, both in the presence or absence of S9-mix in two independent experiments. No increase in the mean number of revertants per plate was observed in any tester strain at the indicated concentrations. The positive and negative controls included for each tester strain showed the expected results. Based on the study results, the test substance was considered to be non-mutagenic in the in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

CAS No. 91052-54-9

The potential mutagenicity of Glycerides, C16-18 mono-, di- and tri was assessed in three S. typhimurium strains (TA 97, TA 98 and TA 100) in an Ames test similar to OECD Guideline No. 471 and under GLP conditions (Kennelly, 1987). In a preliminary test, no cytotoxicity occurred up to 5000 µg/plate. Precipitation of the test substance was observed at 1000 and 5000 µg/plate. However, precipitation did not interfere with the colony counting at 1000 µg/plate. Thus, 1000 µg/plate was chosen as the highest concentration for the main experiment. No cytotoxicity was observed up to the highest, precipitating dose. The frequency of revertant colonies was not increased in any of the bacterial strains tested. The positive and negative controls included in the assay demonstrated the efficiency of the assay. Based on the results of this study, the test substance did not induce mutations in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

ELINCS 460-300-6

A bacterial gene mutation assay with Glycerides, C16-18 and C18-unsaturated, branched and linear mono-, di- and tri- was performed in compliance with OECD guideline 471 and GLP (Bowles, 2004). In a preliminary cytotoxicity test, no cytotoxicity was observed in TA 100 at up to 5000 µg/plate with and without metabolic activation system (S9-mix). Thus, concentrations up to 5000 µg/plate were applied for treatment of S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 in two independent experiments in the presence or absence of metabolic activation. An oily precipitate was observed at 5000 µg/plate (with and without S9-mix) in all experiments, but this did not interfere with the scoring of revertant colonies. The mean number of revertant colonies per plate was not increased in any tester strain and at any test concentration with or without metabolic activation. The mean numbers of spontaneous revertants in the negative (solvent) controls and the mean number of revertants in the positive controls of the strains used were all within the normal historical ranges. Therefore, the test substance was considered to be non-mutagenic in the selected strains of S. typhimurium under the conditions of this Ames test.

CAS No. 65381-09-1

Decanoic acid, ester with 1,2,3-propanetriol octanoate was tested in a bacterial mutation assay according to OECD 471 and GLP (Banduhn, 1989). In two independent experiments, S. typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 were exposed to concentrations ranging from 8 to 5000 µg/plate (first experiment) and 250 to 4000 µg/plate (second experiment), respectively. Both experiments were performed in the absence or presence of a liver microsomal activation system (S9-mix). No cytotoxicity was noted in the treated strains when compared to controls. Significant and dose-related increases in the mean number of revertants per plate in the presence and absence of S9 mix compared to controls were observed in TA 100 at concentrations ≥ 500 µg/plate. However, they did not fulfil the criteria for an overall positive mutagenic result, since the tester strain TA 1535, which covers the same mutation spectrum as TA 100, consistently showed negative results with and without S9 mix in both experiments. Thus, the positive result in TA 100 was not considered to be of biological relevance. The positive control substances for each tester strain showed the expected results and all negative controls were within the historical range of the respective strains, thereby confirming the validity of the study. Under conditions of this assay, the test substance was found to be non-mutagenic in S. typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 both in the absence of presence of S9-mix.

CAS No. 73398-61-5

In a GLP-conform study performed according to EU method B.13/14, the mutagenicity of Triglycerides, mixed decanoyl and octanoyl in bacteria was investigated (Schöberl, 1994). S. typhimurium TA 98, TA 100, TA 1535, TA 1537 and TA 1538 were tested in two independent experiments at concentrations from 8 to 5000 µg/plate both in the presence or absence of metabolic activation (phenobarbiturate-induced rat liver microsomes). No cytotoxicity was observed in any of the tester strains up 5000 µg/plate, but precipitation of the test substance was observed at this concentration both in the presence of absence of the metabolic activation system. Exposure to the test substance in the presence or absence of metabolic activation did not increase the mean number of revertants in any strain of bacteria tested. The positive and negative controls included in the assay yielded the expected results. Under the conditions of this bacterial mutation assay, the test substance was found to be non-mutagenic in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

CAS No. 67701-26-2

The potential mutagenicity of C12-C18 trialkyl glyceride was investigated in a bacterial mutation assay performed similarly to OECD guideline 471 and under GLP conditions (Kennelly, 1987). S. typhimurium TA 97, TA 98 and TA 100 were tested in the presence or absence of a liver metabolic activation system (S9-mix). In a preliminary experiment, the Salmonella strain TA 100 was exposed to concentrations of 8-5000 µg/plate. No cytotoxicity occurred up to 5000 µg/plate, but precipitation of the test substance was observed at this test concentration. Although limited solubility was also observed at 1000 µg/plate, precipitation of the test substance at this concentration did not interfere with colony counting. Thus, 1000 µg/plate was selected as maximum concentration in the main experiment. No increase in the mean number of revertants was observed in any tester strain compared to controls in the presence or absence of metabolic activation. The positive and negative controls included in the assay showed the expected results. Based on the results of this experiment, the test substance was considered to be non-mutagenic in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

CAS No. 97593-30-1

A bacterial gene mutation assay with Glycerol tristearate was performed in compliance with OECD guideline 471 and GLP (Herbold, 2007). In two independent experiments, S. typhimurium TA 98, TA 100, TA 102, TA 1535 and TA 1537 were exposed to concentrations of 50-5000 µg/plate in the presence or absence of metabolic activation system (S9-mix). In the first experiment cytotoxic effects were evident in the presence of S9-mix at ≥ 1581 (TA 100 and TA 1537) and at 5000 µg/plate (TA 1535). In the second experiment, cytotoxicity was observed in all strains at ≥ 1581 µg/plate (with S9-mix), except for TA 1537 which showed cytotoxicity already at ≥ 500 µg/plate in the presence of S9-mix and at 5000 µg/plate in the absence of metabolic activation. Precipitation of the test substance was noted at ≥ 1581 in the first experiment and at 5000 µg/plate in the second experiment, respectively. No increase in the mean number of revertants was observed in any tester strain in the two independent experiments independent of the metabolic activation. The negative (solvent) controls were within the historical range and the positive controls showed the expected results. Based on the results of both experiments, the test substance did not induce mutagenic effects in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

CAS No. 91052-13-0

A bacterial gene mutation assay with Glycerides, C8-18 and C18-unsatd. mono- and di-, acetates was performed according to OECD guideline 471 and GLP (Sarada, 2010). Two independent experiments were performed both in the presence or absence of metabolic activation in S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 and in E. coli WP2 uvrA. In the preliminary toxicity screening, growth inhibitory effects were observed at ≥ 20 µg/plate in S. typhimurium TA 98 and TA 1537 (without metabolic activation), at ≥ 78 µg/plate in S. typhimurium TA 100 and TA 1535 (without metabolic activation), and at ≥ 313 µg/plate in all S. typhimurium strains with metabolic activation. Based on these results, concentrations ranging from 0.61 to 78 µg/plate were used for the tester strains TA 100, TA 1535, TA98 and TA 1537 in the absence of metabolic activation, whereas concentrations ranging from 10 to 313 µg/plate were applied for treatment of the tester strains TA 100, TA 1535, TA 98 and TA 1537 in the presence of metabolic activation. Since no cytotoxicity was seen in E. coli WP2 uvrA, the maximum test concentration of 5000 µg/plate and concentrations of 2500, 1250, 625 and313 µg/plate were selected for treatment in the main assay. Precipitation of the test substance was observed on the plates with E. coli WP2 uvrA at test concentrations ≥ 1250 µg/plate without metabolic activation and at ≥ 2500 µg/plate with metabolic activation in both experiments. No increase in mean revertant number was observed in any bacterial strain after exposure to the test substance in the presence or absence of metabolic activation. The positive and negative controls revealed the expected results. Under the conditions of this assay, the test substance did not induce gene mutations in the selected strains of S. typhimurium and in E. coli WP2 uvrA in the absence and presence of metabolic activation, respectively.

CAS No. 736150-63-3

The potential mutagenicity of Glycerides, castor-oil.mono, hydrogenated, acetates was investigated in a bacterial mutation assay according to OECD guideline 471 and in compliance with GLP (Edwards, 2004). S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102 were exposed to concentrations ranging from 50 to 5000 µg/plate with or without S9-mix in two independent experiments. In both experiments, no dose-dependent increase in the mean number of revertants per plate was observed in any tester strain up to the maximum concentration. Slight cytotoxicity, as indicated by a small reduction in the number of revertants compared to controls, was induced in TA 100 at 5000 µg/plate (with S9-mix) in the preincubation assay and in TA 1535 at 160 µg, 1600 µg and 5000 µg/plate (with S9-mix) in the plate incorporation assay. The solvent and positive control values were within the historical control values. Based on the results of this study, the test substance did not induce mutagenicity in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

Short-, medium- and long-chain triglycerides (SCT, MCT, LCT)

To assess the mutagenic effects in bacteria of medium- and long-chain triacylglycerol oil produced from medium-chain triacylglycerols, S. typhimurium TA98, TA100, TA1535, and TA1537 and E. coli WP2 uvrA were investigated similarly to OECD Guideline 471 (Matulka et al., 2006). In two independent experiments, the tester strains were exposed to concentrations of 313-5000 µg/plate with or without metabolic activation system (S9-mix). No increase in the mean number of revertant colonies per plate was observed in any strain at the indicated concentrations. No cytotoxicity was observed in the experiments up to the limit concentration of 5000 µg/plate. The positive and negative controls included for each tester strains revealed the expected results. Based on the study results, the test substance was considered to have no mutagenic activity in the selected S. typhimurium and E. coli strains with and without metabolic activation.

CAS No. 56-81-5

A bacterial gene mutation assay with Glycerol was performed similarly to OECD guideline 471 (Doolittle, 1988). In one experiment, S. typhimurium TA 98, TA 100, TA 1535, TA1537 and TA 1538 were exposed to concentrations of 200-1000 µg/plate in the presence and absence of metabolic activation system (S9-mix). No cytotoxic effects were observed in any tester strain with and without S9-mix up to the highest concentration tested. In tester strain TA 100, the number of revertants per plate was above the solvent control. However, the increase in the number of revertants was only slight and without concentration relationship. Therefore, the test was repeated with TA 100 using slightly higher glycerol concentrations than in the main experiment, yielding a negative result. No increase in the mean number of revertants was observed in any of the other tester strains in the absence and presence of metabolic activation. The negative (solvent) and the positive controls showed the expected results. Based on the results the study, the test substance did not induce mutagenic effects in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

CAS No. 111-14-8

The potential mutagenicity of heptanoic acid was studied in a bacterial mutation assay performed similarly to OECD guideline 471 (NTP 1987 and Zeiger et al., 1992). S typhimurium TA 1535, TA 1537, TA 97, TA 98 and TA 100 were treated with concentrations ranging from 10 to 6666 µg/plate in the absence and presence of metabolic activation (induced male Sprague Dawley rat and Syrian hamster liver S9 mix (10% and 30%)). No increase in the mean number of revertant colonies per plate was observed in any strain at any test concentrations. In the presence of 30% hamster or 30% rat S9 mix, precipitation occurred at concentrations ≥ 1000 µg/plate in Salmonella strains TA 100, TA 1535, TA 1537 and TA 98 and at 1000 µg/plate in TA 97. In the presence of 10% hamster or 10% rat S9 mix, precipitation occurred at concentrations at 3333 µg/plate in Salmonella strains TA 100, TA 1535, and TA 98 and at 1666 µg/plate in TA 97. However, none of the precipitates interfered with the counting of revertant colonies in any of the tester strains. Cytotoxicity in the bacterial strains was observed starting from 333 µg/plate in TA 97 and from 1000 µg/plate in the other S. typhimurium strains. The positive and negative (solvent) controls included showed the expected results. Under the conditions of this experiment, the test substance did not induce mutagenicity in the selected strains of S. typhimurium in the presence and absence of metabolic activation.

CAS No. 112-85-6

In a GLP- study performed according to OECD guideline 471, the potential mutagenicity of docosanoic acid was investigated (Nakajima, 2002) in two independent experiments. S. typhimurium TA 1535, TA 1537, TA 98 and TA 100 and E. coli WP2 uvrA were exposed to concentrations ranging from 156 to 5000 µg/plate. Precipitation was observed in all test concentrations and in both experiments. No effects on the mean number of revertants were observed compared to controls. The positive and negative controls showed the expected results and thus confirmed the efficiency of the assay. Based on these results, the test substance did not induce mutagenicity in the selected strains of S. typhimurium and E. coli in the presence and absence of metabolic activation.

Genetic toxicity (cytogenicity) in mammalian cells in vitro

CAS No. 111-03-5

2,3-Dihydroxypropyl oleate was assessed in an in vitro mammalian chromosome aberration test in Chinese hamster lung cells (CHL/IU) according to OECD guideline 473 and under GLP conditions (Nakajima, 2005). In a preliminary cytotoxicity test, cells were exposed to concentrations ranging from 6.96 to 3565 µg/mL for a continuous 24-h exposure period with metabolic activation (S9-mix) or a short-term 6 h exposure with and without S9-mix. The test substance was not cytotoxic after short-term exposure at any concentration in the presence of S9 mix. Without metabolic activation, relative growth was reduced to about 71% of the control value after a period of 6 h. Continuous exposure for 24 h without S9-mix caused no cytotoxicity up to concentrations of 1783 µg/mL. At 3565 µg/mL, relative growth was decreased to ca. 13 % of the control value. The concentration leading to 50% cytotoxicity was calculated to be 2738 µg/mL. Based on these results, concentrations of 891, 1783 and 3565 µg/mL (with and without S9 mix) were selected for chromosome analysis after short-term exposure, whereas concentrations of 55.7, 111, 223 and 446 µg/mL (without S9-mix) were chosen for chromosome analysis after continuous exposure. No increase in the number of cells with chromosomal aberrations was observed compared to controls in any of the experiments performed. No cytotoxic effects were observed after short-term exposure, but the test substance was cytotoxic at 446 µg/mL after 24-h continuous treatment. Visible precipitation of the test substance was observed at concentrations ≥ 223 µg/mL, which however did not interfere with chromosomal analysis. The positive controls included during short-term and continuous exposure showed the expected results. Under the conditions of this experiment, the test substance was considered to be not clastogenic in Chinese hamster lung cells (CHL/IU) in the presence and absence of metabolic activation.

CAS No. 8001-79-4

The clastogenic activity of Castor oil was investigated in an in vitro mammalian chromosome aberration test performed similar to OECD guideline 473 and under GLP conditions (Irwin, 1992). Chinese hamster Ovary (CHO) cells were treated with the test substance at 1600, 3000 and 5000 µg/mL for 10 h (without metabolic activation) or 2 h (with metabolic activation), respectively. No increase in the number of cells with chromosomal aberrations was observed at any concentration compared to controls. The positive controls showed the expected increase in the number of cells with chromosomal aberrations. Under the conditions of this assay, the test substance was not clastogenic in Chinese hamster Ovary (CHO) cells in the presence and absence of metabolic activation.

Furthermore, an in vitro Sister Chromatid Exchange Assay was performed with Castor oil in Chinese hamster Ovary (CHO) cells (Irwin, 1992). The occurrence of sister chromatid exchanges was investigated in the presence and absence of metabolic activation (rat liver S9-mix) with test substance concentrations of 0, 160, 500, 1600 and 5000 µg/mL diluted with DMSO. Castor oil did not induce a significant increase in the number of sister chromatid exchanges at any dose level. No relevant cytotoxic effects were reported. Positive controls significantly increased the rate of sister chromatid exchanges indicating the sensitivity of the assay. Based on the results of this assay, the test substance was not considered to be able to induce DNA damage in Chinese hamster Ovary (CHO) cells.

CAS No. 91052-13-0

The clastogenic potential of Glycerides, C8-18 and C18-unsatd. mono- and di-, acetates was assessed in a GLP-study in Chinese hamster lung cells (CHL/IU) according to OECD guideline 473 (Seki, 2010). In a preliminary cell growth inhibition test with concentrations ranging from 9.8 to 5000 µg/plate, no significant inhibition of cell growth was observed after 4 h exposure in the presence and absence of metabolic activation (S9 mix). However, a moderate reduction of cell growth of ca. 30-40% compared to controls was observed at concentrations ranging from 156-1250 µg/mL after 24 and 48 h continuous exposure. Based on the results of this study, concentrations of 20, 39, 78 and 156 µg/mL (± S9 mix) were used for the analysis of chromosomal aberrations after short-term exposure (6 h), whereas concentrations of 78, 156, 313, 625, 1250, 2500 and 5000 µg/mL (± S9 mix) were chosen for analysis chromosomal aberrations after continuous exposure (24 and 48 h). No increase in the number of cells with chromosomal aberrations was observed compared to controls in any of the experiments performed. No cytotoxic effects were observed in any of the experiments performed. An oily precipitation of the test substance was observed at concentrations ≥ 78 µg/mL, but did not interfere with chromosomal analysis of the cells. The positive controls included during short-term and continuous exposure showed the expected results. Under the conditions of this experiment, the test substance was considered to be not clastogenic in Chinese hamster lung cells (CHL/IU) in the presence and absence of metabolic activation.

CAS No. 736150-63-3

An in vitro mammalian chromosome aberration test was performed with Glycerides, castor-oil.mono, hydrogenated, acetates in human lymphocytes according to OECD guideline 473 and in compliance with GLP (Edwards, 2004). An initial cytotoxicity test was performed with concentrations of 313, 625, 1250, 2500 and 5000 µg/mL in the presence or absence of metabolic activation (S9-mix) for of 3 h. A dose-related toxicity was observed. At the highest concentration (5000 µg/mL), the mitotic index was reduced to 64% of the vehicle control without metabolic activation and to 10% with metabolic activation, respectively. Based on these results, concentrations of 625, 1250, 2500 and 5000 µg/mL (without S9-mix) and 625, 1250, 2500, 3600 and 5000 µg/mL (with 2% S9-mix) were chosen for treatment of cells in the main assay for an exposure period of 3 h. Metaphase analysis was performed at concentrations of 1250, 2500 and 5000 µg/mL (without S9-mix) and 625, 1250 and 2500 µg/mL (with 2% S9-mix). In the second main assay, cells were exposed to concentrations of 313-5000 µg/mL (without S9 mix) or 625-5000 µg/mL (with 4% S9 mix) for either 3 or 20 h, respectively. As the samples without metabolic activation revealed mean mitotic indices lower than 50% of the solvent control in all dose groups, this part of the experiment was repeated with lower concentrations ranging from 2.5 to 320 µg/mL. Based on the cytotoxicity data obtained, concentrations of 40, 80 and 160 µg/mL (without S9) and 625, 1250 and 2500 µg/mL (with 4% S9-mix) were used for metaphase analysis. The chromosome analysis of all experiments showed no treatment-related increase in the number of cells with chromosomal aberrations compared to controls. The frequency of metaphases with chromosomal aberrations in the solvent controls was compatible to the historical control values and the positive controls produced statistically significant increases in the frequency of metaphases with chromosomal aberrations. Based on the results of this chromosome aberration test, the test substance was not clastogenic in human lymphocytes in the presence or absence of metabolic activation under the experimental conditions chosen.

Short-, medium- and long-chain triglycerides (SCT, MCT, LCT)

An in vitro mammalian chromosome aberration test with the SALATRIM (short- and long-chain acyl triglyceride molecules) family of triacylglycerols was performed similar to OECD guideline 476 in Chinese hamster Ovary (CHO) cells (Hayes et al., 1994). In a preliminary toxicity test, concentrations of 8.0, 40.0, 200.0, 500.0, and 1000 μg/mL were used to determine suitable concentrations for chromosome analysis. Based on these results, the occurrence of chromosome aberrations was investigated in the presence and absence of metabolic activation (rat liver S9-mix) with test substance concentrations of 250, 500 and 1000 µg/mL and after a harvest time of 8-10 h. The high dose was limited for evaluation due to the low solubility of the fats in the assay medium. No significant increase in the number of phases with aberrations was observed in treated CHO cells at any preparation time and concentration. No significant cytotoxic effects were reported. The positive controls significantly increased the rate of chromosome aberrations. In conclusion, the test substance was not clastogenic in Chinese hamster ovary cells, neither in the presence nor in the absence of a metabolic activation system, under the experimental conditions chosen.

CAS No. 56-81-5

The clastogenic activity of Glycerol was investigated in an in vitro mammalian chromosome aberration test performed similarly to OECD guideline 473 (Doolittle, 1988). Chinese hamster Ovary (CHO) cells were treated with the test substance at 100, 200, 400, 600, 800 and 1000 µg/mL for a period of 10 and 14 h (without metabolic activation) or 2 h (with metabolic activation) followed by a 10 and 14 h recovery period, respectively. After treatment, no cytotoxicity was observed at any test concentration compared to controls. A statistically significant increase in the frequency of cells with chromosomal aberrations was observed in cells treated with 200 µg/mL for2 h in the presence of S9, followed by a 10 h recovery period. Due to the lack of any concentration-response relationship, this isolated result was considered to be spurious and not biologically significant. No increase in the number of metaphases with aberrations was observed at any other preparation time and concentration. The positive controls showed the expected increase in the number of cells with chromosomal aberrations. Under the conditions of this assay, the test substance was not clastogenic in Chinese hamster Ovary (CHO) cells in the presence or absence of metabolic activation.

CAS No. 111-14-8

The clastogenic potential of heptanoic acid was assessed in an in vitro mammalian chromosome aberration test in cultured human lymphocytes performed according to OECD guideline 473 and under GLP conditions (Sarlang, 2010). The study was performed in two independent experiments, both in the presence and in the absence of metabolic activation. In the first experiment, cells were treated at concentrations ranging from 0.16 to 10 mM for a period of 3 h (short-term-treatment). No significant reduction in mitotic index was observed at any concentration in the absence of S9 mix, whereas in the presence of S9 mix a moderate cytotoxicity was observed at 10mM, as shown by a 52% decrease in mitotic index compared to controls. Based on these results, concentrations of 2.5, 5 and 10 mM were selected for chromosome analysis in the presence and absence of metabolic activation. In the second experiment, short-term treatment for 3 h with S9 mix and continuous treatment for 20 and 44 h without S9 mix was performed with the test substance at concentrations ranging from 0.63 to 10 mM. Short-term treatment for 3 h with S9 mix resulted in a moderate decrease in mitotic index (53%) at 10 mM. Therefore, concentrations of 5, 7.5 and 10 mM were chosen for chromosome analysis. After the 20-h treatment in the absence of S9 mix, a 50% reduction in mitotic index compared to control was observed at 2.5 µM, which was selected as the highest concentration for chromosome analysis, followed by concentrations of 1.25 and 0.63 µM. Based on the cytotoxicity observed after continuous treatment for 44 h, 2.5 mM and 10 mM were chosen as only concentrations for chromosome analysis. No increase in the number of cells with chromosomal aberrations was observed compared to controls at any test concentration and treatment duration. The positive controls included during short-term and continuous exposure showed the expected results. Under the conditions of this experiment, the test substance was not clastogenic in cultured human lymphocytes in the presence and absence of metabolic activation.

CAS No. 112-85-6

Docosanoic acid was assayed in an in vitro mammalian chromosome aberration test conducted in accordance with GLP and similarly to OECD guideline 473 (Nakajima, 2002). In the first experiment, Chinese hamster lung (CHL) cells were treated with the test substance at 272 to 3500 µg/mL, with and without metabolic activation (S9-mix) for 6 h (short-term exposure). These concentrations were selected based on a preliminary cytotoxicity test, in which no toxicity was observed up to the highest concentration of 5000 µg/mL. Since no cytotoxicity was observed after 6 h, cells were exposed to 272-3500 µg/mL for a period of 24 and 48 h (continuous exposure) without metabolic activation, which resulted in 50% cytotoxicity at concentrations of 2703 μg/mL (24 h exposure) and 2248 μg/mL (48 h exposure), respectively. Based on these results, concentrations of 875, 1750 and 3500 µg/mL were selected for chromosome analysis in the absence and presence of metabolic activation in the short-term exposure experiment, whereas concentrations of 350, 700, 1400 and 2800 µg/mL (24 h) as well as 288, 575, 1150, 2300 µg/mL (48 h) were chosen in the experiments with continuous exposure. No increase in the number of cells with chromosomal aberrations was observed in treated cells compared to controls in any of the experiments performed. The positive control substance included during continuous exposure yielded the expected results and fell within the range of the historical positive control data. In contrast, the positive control for the short-term exposure failed to induce a significant increase in the number of cells with chromosomal aberrations. However, based on the negative results obtained in all treated cells in this assay, it was concluded that the test substance did not show clastogenic activity in Chinese hamster lung (CHL) cells in the presence and absence of metabolic activation.

Genetic toxicity (mutagenicity) in mammalian cells in vitro

CAS No. 736150-63-3

An in vitro mammalian cell gene mutation assay was performed with Glycerides, castor-oil.mono, hydrogenated, acetates according to OECD guideline 476 and under GLP conditions (Edwards, 2002). In the first experiment, mutations at theTK locusof mouse-lymphoma L5178Y cells were investigated at concentrations of 625, 1250, 2500, 3600 and 5000 µg/mL. In this experiment, cells were exposed to the test material for a period of 3 h in the presence and for 4 h in the absence of metabolic activation (S9-mix). At 3600 µg/mL, the relative total growth was 10-20% compared to the negative controls, thus providing an appropriate maximum concentration for treatment in the second experiment. In this experiment, cells were exposed without S9-mix to concentrations ranging from 313 to 3600 µg/ for a period of 24 h, whereas in the presence of S9-mix, cells were treated with concentrations of 156-3600 µg/mL for a period of 4 h. Since the relative growth with S9-mix was very low (0-2%) at all test concentrations, the 24-h treatment of cells in the absence of S9-mix was repeated with concentrations ranging from 2.5-320 µg/mL, which resulted in appropriate levels of cytotoxicity (10-20% relative growth) at 160 µg/mL. In the presence of metabolic activation, the relative total growth was 9% at 2500 µg/mL in the first experiment and 37 and 0% at 2500 and 3600 µg/mL in the second experiment, respectively. After a 3-day expression period of the cultures, the resistance to 5-trifluorothymidine (TFT) was determined in all experiments. The test substance did not induce a significant increase in the mutant frequency at any preparation time and dose concentration. The positive controls significantly increased mutant frequency. In conclusion, the test substance did not induce mutations in mouse-lymphoma L5178Y cells, neither in the presence nor in the absence of a metabolic activation system, under these experimental conditions.

Short-, medium- and long-chain triglycerides (SCT, MCT, LCT)

The SALATRIM (short- and long-chain acyl triglyceride molecules) family of triacylglycerols was assessed using a gene mutation assay in cultured mammalian cells (Chinese hamster ovary (CHO-K1) cells) similarly to OECD guideline 476 (Hayes et al., 1994). Gene mutations at the HPRT locus were investigated in the presence and absence of metabolic activation (rat liver S9-mix) with concentrations of 31.25, 62.5, 125, 250, 500 and 1000 µg/mL. The highest concentration was limited by the low solubility of the fats in the assay medium. No significant cytotoxicity was reported. An increase in mutant frequency was not observed at any concentration tested whether with or without metabolic activation. The positive controls significantly increased the mutant frequency. Therefore, it was concluded that under the conditions of the study, the test material was not mutagenic at the HPRT locus of Chinese hamster ovary cells in the absence and presence of metabolic activation.

CAS No. 56-81-5

Glycerol was investigated in a HPRT gene mutation assay in cultured mammalian cells (Chinese hamster ovary (CHO) cells) similarly to OECD guideline 476 (Doolittle, 1988). Gene mutations at the HPRT locus were investigated in the presence and absence of metabolic activation (rat liver S9-mix) with concentrations of 100, 200, 400, 600, 800 and 1000 µg/mL after an exposure period of 5 h. No significant cytotoxicity was reported. An at least threefold increase in mutant frequency compared to controls was observed at 800 and 1000 µg/mL in the absence of metabolic activation. However, this increase was not considered biologically significant and did not fulfil the criteria for a positive response due to the lack of a dose-response relationship. No increase in mutant frequency was noted at any other concentration tested, neither with nor without S9 mix. The positive controls significantly increased the mutant frequency. Therefore, it was concluded that under the conditions of the study, the test material was not mutagenic at the HPRT locus of Chinese hamster ovary cells in the absence and presence of metabolic activation.

CAS No. 111-14-8

An in vitro mammalian cell gene mutation assay with heptanoic acid was performed in mouse-lymphoma L5178Y cells according to OECD guideline 476 (Sarlang, 2010). In a preliminary experiment, the cytotoxicity of the test substance was investigated with six concentrations ranging from 0.02 to 10 mM in the presence and absence of metabolic activation under the same exposure conditions as described in the main experiment. No cytotoxicity was observed up to the highest concentration tested. Based on these results, mutations at the TK locus of mouse-lymphoma L5178Y cells were investigated in two independent experiments at test substance concentrations ranging from 0.313 to 10 mM. In the first experiments cell were exposed for 3 h in the presence and absence of S9 mix, whereas periods of 3 h with S9 mix and 24 h were chosen for treatment without S9 mix in the second experiment. After an expression period of 48 h in the presence of 5-trifluorothymidine (TFT) selective medium, the test substance did not induce a significant increase in the mutant frequency at any test substance concentration in both experiments. Slight to moderate cytotoxicity, as indicated by a reduction in relative total growth of 39-50%, was noted in the first experiment after 3-h treatment with S9 mix at concentrations of 2.5 and 5 mM. In the second experiment, moderate to severe cytotoxicity, as indicated by a reduction in total growth of 53-91%, was observed after 24-h treatment without S9 mix at concentrations ≥ 5mM. No cytotoxic effects were seen at any test concentration after 3-h treatment without S9 mix in the first experiment and after 3-h treatment with S9 mix in the second experiment. The positive controls significantly increased mutant frequency. In conclusion, the test substance did not induce gene mutation in mouse-lymphoma L5178Y cells in the presence and absence of metabolic activation.

Genetic toxicity in vivo

CAS No. 91845-19-1

Glycerides, C16-18 and C18-hydroxy mono- and di- was studied in a Mammalian Erythrocyte Micronucleus Test performed according to OECD Guideline 474 and in compliance with GLP (Wallat, 1985). In a preliminary range-finder toxicity study, 2 male and 2 female CFW 1 mice per group were given a single oral application of the test substance (diluted in arachis oil) at 5000, 7500 and 10000 mg/kg bw. After treatment, a slightly reduced activity and ruffled fur was observed in animals at 7500 mg/kg bw and above persisting up to 20 h post-dose. Based on these results, animals of the main study (7 per sex and dose) received 1000, 5000 and 10000 mg/kg bw or the vehicle alone via gavage. Signs of systemic toxicity in the treated animals of the mid and high dose group included slightly reduced activity and ruffled fur, which disappeared within 20 to 24 h after administration. After a post-exposure period of 24, 48 and 72 h, femoral bone marrow was taken from both femurs in animals treated at 10000 mg/kg bw. To determine the frequency of micro-nucleated erythrocytes, 1000 erythrocytes per animal were scored. No increases in the frequency of micronuclei in polychromatic erythrocytes of the femoral bone marrow of CFW 1 mice exposed to 10000 mg/kg bw were observed. The positive control substance (10 mg/kg bw cyclophosphamide) significantly increased the number of polychromatic erythrocytes with micronuclei in male and female animals. Under the conditions of this experiment, the test substance was negative in the Mammalian Erythrocyte Micronucleus Test in male and female CFW 1 mice.

CAS No. 8001-79-4

An in vivo Mammalian Erythrocyte Micronucleus Test was performed with Castor oil in B6C3F1 mice similar to OECD Guideline 474 (Irwin, 1992). Ten animals per group were treated with test substance concentrations of 0, 0.62, 1.25, 2.50, 5.00, 10.0% (w/w) in the diet by oral feeding for 13 weeks (approx. 0, 917, 2022, 3800, 7823, 15017 mg/kg bw/day). Blood smears were prepared from peripheral blood samples obtained by cardiac puncture of dosed and control animals at the termination of the 13 week study. At least 2000 PCE and 10000 NCE from each animal were scored to determine the frequency of micro-nucleated erythrocytes. No signs of systemic toxicity in any of the treated animals were observed. No increases in the frequency of micronuclei in Peripheral Blood Erythrocytes of B6C3F1 mice exposed to Castor Oil in doses up to approx. 15000 mg/kg bw/day fed for 13 weeks occurred, whereas the positive control substance (0.2% urethane) significantly increased the number of normochromatic and polychromatic erythrocytes with micronuclei in three control animals. Based on these results, the test substance was not considered to be clastogenic in vivo.

Short-, medium- and long-chain triglycerides (SCT, MCT, LCT)

An in vivo Mammalian Erythrocyte Micronucleus Test was performed with the SALATRIM (short- and long-chain acyl triglyceride molecules) family of triacylglycerols in Crl:CD BR VAF/Plus rats similarly to OECD Guideline 474 (Hayes et al., 1994a and b). Groups of 20 male and 20 female animals were exposed to either of two SALATRIM fats or corn oil at 10% (w/w) in the diet (equivalent to 7000 mg/kg bw/day) for at least 13 weeks. Erythrocytes were scored to determine the frequency of micro-nucleated erythrocytes. No signs of systemic toxicity in any of the treated animals were observed. No increases in the frequency of micronuclei in polychromatic erythrocytes of the femoral bone marrow of Crl:CD BR VAF/Plus rats exposed to 7000 mg/kg bw/day SALATRIM occurred. Because these data were collected from a 13-week sub-chronic toxicity study, a positive control for micronuclei formation was not included. Under the conditions of this experiment, the test substance was negative in in the micronucleus assay Crl:CD BR VAF/Plus rats.

Overall conclusion for genetic toxicity

The available data on the genetic toxicity of Glycerides comprise numerous studies on the potential induction of gene mutations in bacteria as well as several in vitro studies on cytogenicity and gene mutations in mammalian cells. Two in vivo cytogenicity studies are available. Available genetic toxicity data from surrogate substances including the hydrolysis products glycerol, heptanoic acid and docosanoic acid (as representative short- and long-chain fatty acids) are likewise taken into account for assessment. Taken together, the results of the available in vitro and in in vivo studies were negative. The results of an Ames test with Glycerol trioctanoate (CAS 538-23-8) were ambiguous due to a positive result in S. typhimurium TA 1535 at high concentrations (≥ 6666 µg/plate) only, thus being of questionable biological relevance.

Based on the available data and following the category approach, all members of the Glycerides category are considered to be not mutagenic in vitro and not clastogenic in vitro and in vivo.


Short description of key information:
In none of these studies mutagenicity in bacteria could be observed.
In none of these studies clastogenic effects in mammalian cells could be observed.
In none of these studies mutagenicity in mammalian cells could be observed.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

According to Article 13 of Regulation (EC) No. 1907/2006 "General Requirements for Generation of Information on Intrinsic Properties of substances", information on intrinsic properties of substances may be generated by means other than tests e.g. from information from structurally related substances (grouping or read-across), provided that conditions set out in Annex XI are met. Annex XI, "General rules for adaptation of this standard testing regime set out in Annexes VII to X” states that “substances whose physicochemical, toxicological and ecotoxicological properties are likely to be similar or follow a regular pattern as a result of structural similarity may be considered as a group, or ‘category’ of substances. This avoids the need to test every substance for every endpoint". Since the group concept is applied to the members of the Glycerides category, data will be generated from data for reference source substance(s) to avoid unnecessary animal testing. Additionally, once the group concept is applied, substances will be classified and labelled on this basis.

Therefore, based on the group concept, the available data on genetic toxicity do not meet the classification criteria according to Regulation (EC) 1272/2008 or Directive 67/548/EEC, and are therefore conclusive but not sufficient for classification.