title: Determination of genotoxic phenylhydrazine agaritine in mushrooms using liquid chromatography-electrospray ionization tandem mass spectrometry.
abstract: A new method with good sensitivity and specificity for detecting and quantifying genotoxic hydrazines, agaritine and 4-(hydroxymethyl)phenylhydrazine (HMPH), was developed using liquid chromatography-electrospray tandem mass spectrometry (MS). Synthetic agaritine and HMPH were structurally assigned by 1H-, 13C- and two-dimensional nuclear magnetic resonance (NMR) analysis (HMBC and HMQC), high-resolution fast-atom bombardment (HR-FAB) MS and time of flight (TOF) MS. The polar molecule agaritine was separated on an ODS column using 0.01% AcOH-MeOH (99:1, v/v) as an eluent with a simple solid-phase extraction cleanup. There were no interference peaks for any of the mushrooms. Agaricus spp. contained 1247 and 2017 microg g-1 agaritine. Other species of mushroom had no agaritine. Recoveries of agaritine from spiked mushroom samples were 60.3-114%. Intra-day precision values were 5.5 and 4.2%, and the inter-day precision values were acceptable (15.0 and 23.0%), as agaritine is unstable. The limit of quantification was 0.003 microg g-1. Even a trace amount of agaritine in mushrooms can, therefore, be determined using this method. We also directly analysed HMPH, an active free hydrazine form of genotoxic agaritine, and obtained direct evidence of its absence from mushrooms. A precursor ion scan confirmed that agaritine derivatives, which could exert similar toxicity, were absent. The results indicate that this specific and sensitive analytical method for detecting and quantifying agaritine and its derivatives could help evaluate the risk of mushroom hydrazines to humans.
authors.affiliation: National Institute of Health Sciences, Division of Foods, Kamiyoga 1-18-1, Setagaya, Tokyo 158-8501, Japan. kondo@nihs.go.jp
authors.forename: Kazunari
authors.initials: K
authors.lastname: Kondo
authors.forename: Asako
authors.initials: A
authors.lastname: Watanabe
authors.forename: Yuko
authors.initials: Y
authors.lastname: Iwanaga
authors.forename: Ikuro
authors.initials: I
authors.lastname: Abe
authors.forename: Hideya
authors.initials: H
authors.lastname: Tanaka
authors.forename: Megumi Hamano
authors.initials: MH
authors.lastname: Nagaoka
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Akiyama
authors.forename: Tamio
authors.initials: T
authors.lastname: Maitani
mesh_terms: D000363:Agaricales / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D010659:Phenylhydrazines / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D016339:Spectrometry, Mass, Fast Atom Bombardment / Q000379:methods*
chemical_list: D010659:Phenylhydrazines; C015654:agaritine