Table D.1.

In vitro and in vivo genotoxicity data for furan‐substituted substances evaluated by the JECFA at the 65th (JECFA, 2006b) and 69th meeting (JECFA, 2009a)

Chemical name [FL‐no:] JECFA‐no.	Test system	Test object	Concentration/dose and test conditions	Results	Reference
In vitro
2‐Methylfuran [13.030]cc 1487	Reverse mutation	S. Typhimurium TA98 and TA100	0.165, 0.330, 0.495 or 0.660 μmol/plate (13.5, 27.1, 40.6 or 54.2 μg/plate)a	Negativeb	Shinohara et al. (1986)
	Reverse mutation	S. Typhimurium TA98, TA100, TA102 and TA1535	Up to 10,000 μg/plate	Negativeb,c,d	Zeiger et al. (1992)
	Reverse mutation	S. Typhimurium TA97 and TA104	Up to 10,000 μg/plate	Equivocalb,c,d	Zeiger et al. (1992)
	Reverse mutation	S. Typhimurium TA98, TA100 and TA102	11 nmol/plate to 1.1 mmol/plate (0.9–90310 μg/plate)a	Negativeb	Aeschbacher et al. (1989)
	DNA damage	B. subtilis H17 (rec+) and M45 (rec−)	0.16, 16 or 1,600 μg/disc	Negative Positiveb,e	Shinohara et al. (1986)
	Chromosomal aberration	CHO cells	0–150 mmol/L (0–12,315 μg/mL)a	Positiveb,f	Stich et al. (1981)
2,5‐Dimethylfuran [13.029]cc 1488	Reverse mutation	S. Typhimurium TA98 and TA100	0.165, 0.330, 0.495 or 0.660 μmol/plate (13.5, 27.1, 40.6 or 54.2 μg/plate)g	Negativeb	Shinohara et al. (1986)
	Reverse mutation	S. Typhimurium TA98 and TA100	Not specified	Negativeb	Lee et al. (1994)
	Reverse mutation	S. Typhimurium TA97, TA98, TA100 and TA1535	Up to 3,333 μg/plate	Negativeb,c,d	Zeiger et al. (1992)
	DNA damage	B. subtilis H17 (rec+) and M45 (rec−)	190, 1900 or 9500 μg/disc	Negative Positiveb,h	Shinohara et al. (1986)
	Chromosomal aberration	Chinese hamster V79 cells	1 mmol/L (96.13 μg/mL)g	Negative	Ochi and Ohsawa (1985)
	Chromosomal aberration	CHO cells	0–20 mmol/L (0–1,923 μg/mL)g	Positiveb,f	Stich et al. (1981)
3‐Methyl‐2‐(3‐methylbut‐2‐enyl)‐furan [13.148] 1494	Reverse mutation	S. Typhimurium TA98, TA100, TA1535 and TA1537	3.2, 16, 80, 400 or 2,000 μg/plate	Negativeb	Asquith (1989)
3‐(2‐Furyl)acrolein [13.034]dd	Reverse mutation	S. Typhimurium TA100	Not specified	Negativeb,c	Eder et al. (1991)
	DNA damage (SOS chromotest)	E. coli PQ37	Not specified	Negativei	Eder et al. (1991)
	DNA damage (SOS chromotest)	E. coli PQ37	Not specified	Weakly positivej	Eder et al. (1993)
2‐Acetylfuran [13.054] 150	Reverse mutation	S. Typhimurium TA98 and TA100	0.165, 0.330, 0.495 or 0.660 μmol/plate (13.5, 27.1, 40.6 or 54.2 μg/plate)j	Negative Positiveb,k	Shinohara et al. (1986)
	DNA damage	E. coli PQ37 (SOS chromotest)	Not specified	Slightly positivej	Eder et al. (1993)
	DNA damage	B. subtilis H17 (rec+) and M45 (rec−)	550, 5,500 or 55,000 μg/disc	Negative Positiveb,L	Shinohara et al. (1986)
	Chromosomal aberration	CHO cells	0–112.6 mmol/ L (0–13,220 μg/mL)j	Positiveb,m,n	Stich et al. (1981)
	UDS	Human hepatocytes	2.19, 4.38, 8.75, 17.5, 35, 70, 140 or 280 μg/ mL	Negative	Durward (2007a)
4‐(2‐Furyl)‐3‐buten‐2‐one [13.044]dd 1511	Reverse mutation	S. Typhimurium TA98, TA100, TA1535 and TA1537	33, 100, 333, 1,000, 2,166 or 3,333 μg/plate	Negativeb,c,o	Mortelmans et al. (1986)
Ethyl 3‐(2‐furyl)propanoate [13.022] 1513	Reverse mutation	S. Typhimurium TA98, TA100, TA1535, TA1537 and TA1538	Up to 3,600 μg/plate	Negativeb	Wild et al. (1983)
O‐Ethyl‐S‐(2‐furylmethyl)thio‐ carbonate [13.191]ee 1526	Reverse mutation	S. Typhimurium TA98, TA100, TA1535 and TA1537	33, 100, 333, 1,000 or 3,330 μg/plate	Negativeb,p	Verspeek‐Rip (2000)
	Reverse mutation	E. coli WP2uvrA	33, 100, 333, 1,000 or 3,330 μg/plate	Negativeb,q	Verspeek‐Rip (2000)
	Chromosomal aberration	Human peripheral lymphocytes	150, 300 or 350 μg/mL	Negativeb,r	Meerts (2000)
	Chromosomal aberration	Human peripheral lymphocytes	130, 240 or 280 μg/mL	Positivei,s	Meerts (2000)
	Chromosomal aberration	Human peripheral lymphocytes	100, 130 or 240 μg/mL	Positivei,t	Meerts (2000)
	Chromosomal aberration	Human peripheral lymphocytes	150, 325 or 375 μg/mL	Negative Positiver,u,v	Meerts (2000)
In vivo
2‐Methylfuran [13.030]cc 1487	Chromosomal aberration	Mouse bone marrow cells and spermatocytes	1,000, 2,000 or 4,000 mg/kg (100, 200 or 400 mg/kg bw per day)w	Negative	Subramanyam et al. (1989)
2‐Acetylfuran [13.054] 1503	Chromosomal aberration	Mouse bone marrow	1,000, 2,000 or 3,000 mg/ L (20, 40 or 60 mg/kg bw)x	Positivey,z	Sujatha et al. (1993)
	Chromosomal aberration	Mouse spermatocytes	1,000, 2,000 or 3,000 mg/ L (20, 40 or 60 mg/kg bw)x	Negativeaa	Sujatha et al. (1993)
	SCE	Mouse bone marrow	1,000, 2,000 or 3,000 mg/ L (20, 40 or 60 mg/kg bw)x	Positive	Sujatha (2007)
	UDS	Rat liver	7 or 21 mg/kg bw	Negative	Durward (2007b)
O‐Ethyl‐S‐(2‐furylmethyl)thio‐carbonate [13.191]ee 1526	Micronucleus induction	Mouse bone marrow	100, 250 or 500 mg/kg bwbb	Negative	Verspeek‐Rip (2001)

CHO: Chinese hamster ovary; SCE: sister chromatid exchange; UDS: unscheduled DNA synthesis.

^aCalculated using relative molecular mass of 2‐methylfuran = 82.1.

^bWith and without metabolic activation.

^cPre‐incubation method.

^dOccasional incidences of slight to complete clearing of the background lawn at the higher concentrations.

^eNegative at all concentrations with metabolic activation; positive without metabolic activation.

^fClastogenic activity decreased with metabolic activation (statistical significance of results was not specified).

^gCalculated using relative molecular mass of 2,5‐dimethylfuran = 96.13.

^hPositive at every concentration without metabolic activation; with metabolic activation, negative at 190 μg/disc, but positive at higher concentrations.

ⁱWithout metabolic activation.

^jCalculated using relative molecular mass of 2‐furyl methyl ketone = 110.11.

^kPositive only in strain TA98 with an increase in the presence of metabolic activation.

^lNegative at 550 μg/disc; positive at 5,500 and 55,000 μg/disc (with and without metabolic activation).

^mCytotoxicity was observed at 12,398 μg/ mL (112.6 mmol/ L) in the presence of metabolic activation.

ⁿClastogenic activity increased with metabolic activation (statistical significance of results was not specified).

^oCytotoxicity was observed at 3,333 μg/plate in all S. typhimurium strains and at 2166 μg/plate in S. typhimurium strains TA100 and TA1537.

^pCytotoxicity was observed at the 3,330 μg/plate level in all S. typhimurium strains and at 1,000 μg/plate in S. typhimurium strains TA100 and TA1535.

^qCytotoxicity was observed at 3,330 μg/plate in the absence of metabolic activation.

^r3‐h continuous exposure time.

^s24‐h continuous exposure time.

^t48‐h continuous exposure time.

^uWith metabolic activation.

^vStatistically significant dose‐dependent increases in chromosomal aberrations were seen at the two highest concentrations only, 325 and 375 μg/ mL.

^wMice received 2‐methylfuran in the diet for 5 consecutive days at 24‐h intervals.

^xTwo experimental protocols were utilised. In one experiment, animals received single oral dose administrations of the test compound. In the other experiment, the test compound was orally administered once per day at the same concentrations as in the single‐dose study for 5 consecutive days with 24‐h intervals between doses.

^yNo effects observed at 20 mg/kg bw dose level and only mild, but significant (p < 0.05) effects seen at higher concentrations in bone marrow cells.

^zChromosomal aberrations were observed in the presence of significant mitodepression.

^aaA single statistically significant occurrence of increased chromosomal aberrations observed 3 weeks following a single dose administration in the 60 mg/kg bw test group; statistically significant increases in polyploidy and XY univalents observed at weeks 3 and 4 at 60 mg/kg bw in multipledose‐treated rats.

^bbSingle dose administered by gavage.

^ccSubstance deleted from the Union list. Commission Regulation (EU) No 246/2014 of 13 March 2014 amending Annex I to Regulation (EC) No 1334/2008 of the European Parliament and of the Council as regards removal from the Union list of certain flavouring substances. OJ L74, 14.3.2014, p. 58–60.

^ddSubstance not supported by industry (DG SANTE, 2019).

^eeSubstance not supported by industry and not included in the Union List. Commission Implementing Regulation (EU) No 872/2012 of 1 October 2012 adopting the list of flavouring substances provided for by Regulation (EC) No 2232/96 of the European Parliament and of the Council, introducing it in Annex I to Regulation (EC) No 1334/2008 of the European Parliament and of the Council and repealing Commission Regulation (EC) No 1565/2000 and Commission Decision 1999/217/EC. OJ L 267, 2.10.2012, p. 1–161.