title: A simple and highly sensitive HPLC method with fluorescent detection for determination of pipecolic acid in mouse brain areas.
abstract: A simple and highly sensitive high-performance liquid chromatography method for the determination of pipecolic acid in mouse brain areas was developed. After homogenization of brain and pretreatment with o-phthalaldehyde, the pipecolic acid and (2S,3S)-3-methylpyrrolidine-2-carboxylic acid (internal standard) were derivatized with 4-(5,6-dimethoxy-2-phthalimidinyl)-2-methoxyphenylsulfonyl chloride at 70 °C for 15 min in basic medium (pH 9.0). The fluorescent derivatives of pipecolic acid and internal standard were separated on a reversed-phase column by stepwise elution using acetic acid (30 mM)-acetonitrile at 50 °C and detected by fluorescence measurement at 316 nm (excitation) and 403 nm (emission). The detection limit (signal-to-noise ratio=3) of pipecolic acid was 13 fmol per injection. The recovery was about 106.7%. The precision (relative standard deviation) was 3.2%.
authors.affiliation: Faculty of Pharmacy and Pharmaceutical Sciences, Fukuyama University, Fukuyama, Hiroshima 729–0292, Japan. inoue@fupharm.fukuyama-u.ac.jp
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Inoue
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Sakata
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nishio
authors.forename: Kohji
authors.initials: K
authors.lastname: Tokumo
authors.forename: Eijiro
authors.initials: E
authors.lastname: Kojima
authors.forename: Yuuko
authors.initials: Y
authors.lastname: Date
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Tamura
authors.forename: Yasuto
authors.initials: Y
authors.lastname: Tsuruta
mesh_terms: D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005453:Fluorescence; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D010875:Pipecolic Acids / Q000032:analysis*
chemical_list: D010875:Pipecolic Acids; C031345:pipecolic acid