title: Development and application of electrospray-active derivatization reagents for hydroxysteroids.
abstract: New derivatization reagents, 1-(2,4-dinitro-5-fluorophenyl)-4-methylpiperazine (PPZ) and 4-(4-methyl-1-piperazyl)-3-nitrobenzoyl azide (APZ), were developed for the liquid chromatography-electrospray ionization-mass spectrometric (LC-ESI-MS) determination of steroids having a hydroxy group. PPZ reacted with a phenolic hydroxy group in estrogens. After quaternarization of the PPZ-estrogens with methyl iodide, the resulting derivatives provided more than a 2000-fold higher sensitivity compared to the intact estrogens. After derivatization of steroids having an alcoholic hydroxy group (5-ene-steroids or 5alpha-reduced steroids) with APZ followed by methylation, their detection responses increased more than 500 times. These derivatization procedures coupled with LC-ESI-MS/MS were successfully used for the determination of estrogens in the serum and prostatic 5alpha-dihydrotestosterone.
authors.affiliation: Division of Pharmaceutical Sciences, Graduate School of Natural Science and Technology, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Nishio
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Higashi
authors.forename: Anju
authors.initials: A
authors.lastname: Funaishi
authors.forename: Jun
authors.initials: J
authors.lastname: Tanaka
authors.forename: Kazutake
authors.initials: K
authors.lastname: Shimada
mesh_terms: D001386:Azides / Q000138:chemical synthesis / Q000737:chemistry*; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods; D013196:Dihydrotestosterone / Q000097:blood; D004967:Estrogens / Q000097:blood; D005260:Female; D006801:Humans; D006914:Hydroxysteroids / Q000032:analysis* / Q000737:chemistry*; D008297:Male; D008745:Methylation; D015394:Molecular Structure; D010879:Piperazines / Q000138:chemical synthesis / Q000737:chemistry*; D011247:Pregnancy; D011467:Prostate / Q000737:chemistry; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D001386:Azides; D004967:Estrogens; D006914:Hydroxysteroids; D010879:Piperazines; D013196:Dihydrotestosterone