title: Lack of in vivo clastogenic activity of grape seed and grape skin extracts in a mouse micronucleus assay.
abstract: Meganatural brand grape seed extract (GSE) and grape skin extract (GSKE), containing proanthocyanidin polyphenolic compounds, are intended for use in food as functional ingredients exhibiting antioxidant activity. Proanthocyanidins, as well as the minor constituent phenolic compounds in GSE and GSKE, are present naturally in many foods such as fruits, vegetables, chocolate, tea, etc., and on average people consume 460-1000 mg/day of these combined substances. While some polyphenolic compounds, tested individually, have demonstrated antitumorigenic or antipromotional activity, at least one minor component of GSE and GSKE, quercitin, has exhibited positive activity in Salmonella and other in vitro mutagenicity assays. As part of a program to investigate the safety of GSE and GSKE, these products were tested for in vivo clastogenic activity and/or disruption of the mitotic apparatus by detecting micronuclei in polychromatic erythrocyte (PCE) cells in Crl:CD-1(ICR) BR mouse bone marrow. The appropriate test article was dissolved in 0.5% carboxymethylcellulose and dosed by oral gavage to five males/test article/dose level/harvest time point. Animals were dosed at 500, 1000 and 2000 mg/kg. Five animals dosed with either test article at 500, 1000 and 2000 mg/kg dose levels and five animals dosed with the cyclophosphamide (80 mg/kg) positive control were euthanized approximately 24 h after dosing for extraction of bone marrow. Five animals dosed with either test article at the 2000 mg/kg dose level and five animals dosed with the vehicle control article were euthanized approximately 24 and 48 h after dosing for extraction of bone marrow. At least 2000 PCEs per animal were analyzed for frequency of micronuclei. Cytotoxicity was assessed by scoring the number of PCEs and normochromatic erythrocytes (NCEs) in at least the first 500 erythrocytes for each animal. For both GSE and GSKE, no statistically significant increase in micronucleated PCEs was observed at any dose level or harvest time point. GSE produced indication of cytotoxicity (decreased PCE:NCE ratio) at the 2000 mg/kg dose level for the 48-h harvest time point, confirming that the test article reached the target bone marrow in significant amount. Meganatural GSE and Meganatural GSKE were evaluated as negative in the mouse bone marrow micronucleus assay under the conditions of this assay.
authors.affiliation: Covance Laboratories Inc., 9200 Leesburg Pike, Vienna, VA 22182, USA. gregory.erexson@covance.com
authors.forename: G L
authors.initials: GL
authors.lastname: Erexson
mesh_terms: D000284:Administration, Oral; D000818:Animals; D000872:Anthocyanins / Q000008:administration & dosage / Q000633:toxicity; D000975:Antioxidants / Q000008:administration & dosage / Q000633:toxicity; D001854:Bone Marrow Cells / Q000187:drug effects; D002869:Chromosome Aberrations; D002875:Chromosomes / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004912:Erythrocytes / Q000187:drug effects; D005419:Flavonoids; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D015162:Micronucleus Tests / Q000379:methods; D009153:Mutagens / Q000633:toxicity*; D010636:Phenols / Q000008:administration & dosage / Q000633:toxicity; D010936:Plant Extracts / Q000008:administration & dosage / Q000633:toxicity*; D011108:Polymers / Q000008:administration & dosage / Q000633:toxicity; D044945:Proanthocyanidins; D011897:Random Allocation; D012449:Safety; D012639:Seeds / Q000737:chemistry*; D027843:Vitis / Q000737:chemistry*
chemical_list: D000872:Anthocyanins; D000975:Antioxidants; D005419:Flavonoids; D009153:Mutagens; D010636:Phenols; D010936:Plant Extracts; D011108:Polymers; D044945:Proanthocyanidins; C013221:proanthocyanidin