title: Identification of novel electrophilic metabolites of piper methysticum Forst (Kava).
abstract: Dietary supplements containing Piper methysticum Forst. (kava) have been implicated in multiple cases of liver injury in humans, including 10 recently reviewed cases in which patients required liver transplantation following the usage of kava-containing products (Centers for Disease Control and Prevention, reprinted. (2003) J. Am. Med. Assoc. 289, 36-37). To investigate a possible mechanism(s) of kava-induced hepatotoxicity, an extract of kava was incubated in vitro with hepatic microsomes, NADPH, and GSH. Electrophilic intermediates that were generated via metabolic activation were trapped as GSH conjugates and removed from the protein mixture using ultrafiltration. Positive ion electrospray LC-MS/MS with precursor ion scanning was used for the selective detection of GSH conjugates, and LC-MS(n) product ion scanning was used to elucidate their structures. Using this in vitro MS-based screening assay, two novel electrophilic metabolites of kava, 11,12-dihydroxy-7,8-dihydrokavain-o-quinone and 11,12-dihydroxykavain-o-quinone, were identified. Mercapturic acids of these quinoid species were not detected in the urine of a human volunteer following ingestion of a dietary supplement that contained kava; instead, the corresponding catechols were metabolized extensively to glucuronic acid and sulfate conjugates. These observations indicate that quinoid metabolites, under most circumstances, are probably not formed in substantial quantities following the ingestion of moderate doses of kava. However, the formation of electrophilic quinoid metabolites by hepatic microsomes in vitro suggests that such metabolites might contribute to hepatotoxicity in humans when metabolic pathways are altered (e.g., because of a drug interaction, genetic difference in enzyme expression, etc.) or if conjugation pathways become saturated.
authors.affiliation: Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, 833 South Wood Street, Chicago, Illinois 60612-7231, USA.
authors.forename: Benjamin M
authors.initials: BM
authors.lastname: Johnson
authors.forename: Sheng-Xiang
authors.initials: SX
authors.lastname: Qiu
authors.forename: Shide
authors.initials: S
authors.lastname: Zhang
authors.forename: Fagen
authors.initials: F
authors.lastname: Zhang
authors.forename: Joanna E
authors.initials: JE
authors.lastname: Burdette
authors.forename: Linning
authors.initials: L
authors.lastname: Yu
authors.forename: Judy L
authors.initials: JL
authors.lastname: Bolton
authors.forename: Richard B
authors.initials: RB
authors.lastname: van Breemen
mesh_terms: D000111:Acetylcysteine / Q000652:urine; D000328:Adult; D000818:Animals; D019587:Dietary Supplements / Q000009:adverse effects; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D020901:Kava / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D010936:Plant Extracts / Q000378:metabolism* / Q000493:pharmacokinetics / Q000633:toxicity; D051381:Rats; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D010936:Plant Extracts; D005978:Glutathione; D000111:Acetylcysteine