. 2019 May 22;17(5):e05676. doi: 10.2903/j.efsa.2019.5676

Table E.1.

In vitro genotoxicity data evaluated in FGE.210Rev1

Chemical name [FL‐no]	Test system in vitro	Test object	Concentrations of substance and test conditions	Result	Reference	Comments
α‐Ionone [07.007]	Reverse mutation	S. typhimurium TA98, TA100, TA1535, TA1537 and TA102	0.3–5,000 μg/platea ^, c 0.3–5,000 μg/plateb ^, c	Negative Negative	Bowen (2011)	Toxicity was observed at 1,000 and/or 5,000 μg/plate across all strains in the absence and presence of S9‐mix; no clear evidence of toxicity in TA100 in the presence of S9‐mix. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix
		S. typhimurium TA98 TA100	156.3–5,000 μg/plateb ^, d 156.3–5,000 μg/platea ^, c or b ^, d	Negative Negative		Evidence of toxicity was observed at the highest three or four concentrations across all strains in the absence and presence of S9‐mix. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix
		S. typhimurium TA98 TA1535, TA1537 and TA102	78.1–2,500 μg/platea ^, c 78.1–2,500 μg/platea ^, c or b ^, d	Negative Negative		Evidence of toxicity was observed at the highest three or four concentrations across all strains in the absence or presence of S9‐mix. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix
		S. typhimurium TA1535 TA102 TA1537	39.1–2,500 μg/platea ^, c or b ^, d 19.5–1,250 μg/platea ^, c or b ^, d 19.5–1,250 μg/plateb ^, d	Negative		Evidence of toxicity was observed at the highest three or four concentrations across all strains in the absence or presence of S9‐mix. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix
	Micronucleus induction	Human peripheral blood lymphocytes	160–180 μg/mLb ^, h 40–65 μg/mLa ^, i	Negative Negative	Lloyd (2013b)	The MNBN cell frequencies in all treated cultures fell within the normal range. The study does not comply with OECD Guideline 487; therefore, it has limited validity
Allyl α‐ionone [07.061]	Reverse mutation	S. typhimurium TA102	1.6–5,000 μg/platea ^, c or b ^, c	Negative	Ballantyne (2011)	No evidence of toxicity was observed at any concentration. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix
	Reverse mutation		51.2–5,000 μg/platea ^, c or b ^, d	Negative	Ballantyne (2011)	No evidence of toxicity was observed at any concentration. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix
	Reverse mutation	S. typhimurium TA1535, TA100, TA1537, TA1538, TA98	Five concentrations up to cytotoxicity or max 3,600 μg/platea ^, c	Negative	Wild et al. (1983)	Limited validity (no TA102 or E. coli)
	Micronucleus induction	Human peripheral blood lymphocytes	110–160 μg/mLb ^, h 25–38 μg/mLa ^, i	Negative Negative	Lloyd (2013a)	The MNBN cell frequencies in all treated cultures fell within the normal range. The study does not comply with OECD Guideline 487; therefore, it has limited validity
δ‐Damascone [07.130]	Reverse mutation	S. typhimurium TA98, TA100, TA1535 and TA1537	4.9–5,000 μg/plated ^, e	Negative	Shinya (2006)	Evidence of toxicity was observed at the top three or four concentrations tested. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix
		S. typhimurium TA98, TA100, TA1535 and TA1537	2.4–78.1 μg platea ^, d 9.8–313 μg/plateb ^, d	Negative		Evidence of toxicity was observed at the top concentration in all strains in the absence of S9‐mix and at 156 μg/plate or above in the presence of S9‐mix. The study complies with current recommendations for upper concentration limit inclusion. The study included three replicate plates per concentration and was GLP compliant
		E. coli WP2 uvrA	4.9–5,000 μg/platee ^, d	Negative		Evidence of toxicity was observed at the top three or four concentrations tested. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix
		E. coli WP2 uvrA	2.4–78.1 μg/platea ^, d 9.8–313 μg/plateb ^, d	Negative		Evidence of toxicity was observed at the highest concentration in the presence of S9‐mix. The study complies with current recommendations for upper concentration limit inclusion. The study included three replicate plates per concentration and was GLP compliant
α‐Damascone [07.134]	Reverse mutation	S. typhimurium TA98 and TA100	10–5,000 μg/platee ^, c	Negative	Haddouk (2001)	In TA98, slight to marked toxicity was observed at concentrations ≥ 100 μg/plate or 500 μg/plate with and without S9‐mix, respectively. In TA100, toxicity was observed at concentrations ≥ 500 μg/plate with and without S9‐mix
		S. typhimurium TA1537, TA98	7.8–125 μg/platee ^, c 15.6–250 μg/plateb ^, d	Negative Negative		Slight toxicity was observed in all strains. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix.
		S. typhimurium TA100 and TA1535	31.2–500 μg/platee ^, c 31.2 – 500 μg/plateb ^, d	Negative Negative		Slight toxicity was observed in all strains. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix.
		S. typhimurium TA1535, TA1537, TA98 and TA100	15.6–250 μg/platea ^, c	Negative		Slight toxicity was observed in all strains. No statistically significant increase in revertant numbers was seen at any concentration.
		E. coli WP2uvrA	10–5,000 μg/platee ^, c	Negative		Slight toxicity was observed at 2,500 μg/plate and above without S9‐mix.
		E. coli WP2 uvrA	312.5–5,000 μg/platee ^, c 312.5–5,000 μg/platea ^, c or b ^, d	Negative Negative		Slight toxicity was observed only at the highest concentration tested without S9‐mix. No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix
	Micronucleus induction	Human peripheral blood lymphocytes	9–22 μg/mLa ^, h 12–22 μg/mLb ^, h 5–10 μg/mLa ^, i	Negative Weakly positive Negative	Lloyd (2012)	Weakly positive result was obtained only in the 3 + 21 h treatment in the presence of S9‐mix. Study design complies with OECD Guideline 487
			7.5–14 μg/mLb ^, h ^, j 14–20 μg/mLb ^, h 7.5–14 μg/mLb ^, h ^, j	Positive	Whitwell (2012)	Follow‐up study to explore different methods of mixing and sample preparation to overcome the challenges in inconsistent cytotoxicity that results in difficulties in choosing concentrations for scoring of micronucleated binucleate cells. Experiment conducted only for 3 + 21 h in the presence of S9‐mix
			10–18 μg/mLb ^, h 7–14 μg/mLa ^, i	Positive at high toxic concentrations only	Lloyd (2013c)	Positive results were obtained only at high toxic concentrations in both test conditions. Study is robust and complies with GLP
β‐Ionone epoxide [07.170]	Reverse mutation	S. typhimurium TA98, TA100, TA1535 and TA1537	5–500 μg/platee ^, c	Negative	Jones and Wilson (1988)	No statistically significant increase in revertant numbers was seen at any concentration, either in the presence or absence of S9‐mix
		S. typhimurium TA97a, TA98, TA100, TA1535	501, 1,582 and 5,000 μg/platea ^, c 158, 501 and 1,582 μg/plateb ^, c	Negative Negative	Kringstad (2005)	Evidence of toxicity was observed at the highest concentration in strain TA97a in the absence of S9‐mix and in TA100 in the absence and presence of S9‐mix. The study, therefore, complies with current recommendations for upper concentration limit inclusion. The study included three replicate plates per concentration and was GLP compliant
		E. coli WP2uvrA	501, 1,582 and 5,000 μg/platea ^, c 158, 501 and 1,582 μg/plateb ^, c	Negative Negative	Kringstad (2005)
	tk Mutation induction	Mouse Lymphoma L5178Y TK +/− 3.7.2c cells	200–900 μg/mLe ^, f 4.1–520 μg/mLa ^, g	Negative Negative	Flanders (2006)	A preliminary range‐finder assay was conducted to establish maximum concentrations. Top concentrations in each arm of the study induced 77, 85 and 80% reductions in relative total growth. The study, therefore, complies with current recommendations

Without S9 metabolic activation.

With S9 metabolic activation.

Plate incorporation method.

Pre‐incubation with S9 method.

With and without S9 metabolic activation.

4‐h treatment.

24‐h treatment.

3‐h treatment with 21‐h recovery.

ⁱ

24‐h treatment with 0‐h recovery.

Standard treatment in larger than typical vessel.