Investigations into the genotoxic potential of olive extracts.
The phenolic anti-oxidant 3-hydroxytyrosol (HT) is a major constituent of olives and olive oil. Published data showed it was negative in the Ames test at concentrations up to 5 μL per plate, but did induce chromosomal aberrations in human lymphocytes. HIDROX, an olive extract containing approximately 2.4% HT, was reported as both positive and equivocal in an Ames test in different papers from the same laboratory. Negative results for micronucleus induction in vivo in both an acute study and as part of a 90-day rat toxicity study were also reported for HIDROX. Given the widespread use and consumption of olives, olive oil and olive extracts, it was important to obtain more data. Here we confirm that pure HT, and an olive extract containing 15% HT, both induced micronuclei in cultured cells in vitro, but show that these responses were either due to high levels of cytotoxicity or to reaction of HT with culture medium components to produce hydrogen peroxide. Another extract (H40) containing 40% HT also induced micronuclei in vitro, probably via the same mechanism. However, both extracts were negative in robust Ames tests. The 15% HT formulated extract did not induce micronuclei in rat bone marrow after 4 weeks of dosing up to 561 mg HT/kg/day. H40 produced increased rat bone marrow micronucleus frequencies at 250 and 500 mg HT/kg/day in a 90-day toxicity study, but the results were questionable for various reasons. However, when two different batches of this extract were tested in acute micronucleus studies at doses up to 2000 mg HT/kg, giving plasma exposures that exceeded those in the 90-day study, negative results were obtained. Based on weight of evidence it is concluded that the olive extracts tested are not genotoxic at high doses in vivo, and any genotoxic risks for human consumers are negligible.
Kirkland Consulting, PO Box 79, Tadcaster LS24 0AS, United Kingdom. Electronic address: dkirkland@genetoxconsulting.co.uk.
David
D
Kirkland
DSM Nutritional Products AG, NIC-RD/HN Product Safety, Wurmisweg 576, CH-4303 Kaiseraugst, Switzerland.
James
J
Edwards
DSM Nutritional Products AG, NIC-RD/HN Product Safety, Wurmisweg 576, CH-4303 Kaiseraugst, Switzerland.
Tina
T
Woehrle
DSM Nutritional Products AG, NIC-RD/HN Product Safety, Wurmisweg 576, CH-4303 Kaiseraugst, Switzerland.
Paul
P
Beilstein
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016466:CHO Cells; D002869:Chromosome Aberrations / Q000187:drug effects; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry; D004249:DNA Damage; D006719:Homovanillic Acid / Q000097:blood; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis / Q000138:chemical synthesis; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009152:Mutagenicity Tests; D009153:Mutagens / Q000302:isolation & purification / Q000493:pharmacokinetics / Q000633:toxicity*; D031658:Olea / Q000737:chemistry*; D010626:Phenylethyl Alcohol / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000493:pharmacokinetics / Q000633:toxicity; D010936:Plant Extracts / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D051381:Rats
D003470:Culture Media; D009153:Mutagens; D010936:Plant Extracts; C005975:3,4-dihydroxyphenylethanol; D006861:Hydrogen Peroxide; D010626:Phenylethyl Alcohol; D006719:Homovanillic Acid
Ames tests; Chromosomal aberrations; Genotoxicity; Hydroxytyrosol; Micronuclei; Olive extracts