title: Fluorescent high-performance liquid chromatography assay for lipophilic alcohols.
abstract: A new ultrasensitive fluorescent derivatization procedure for chromatographic analysis of primary, secondary, and nonpolar tertiary alcohols is described. The procedure uses Bodipy FL in basic dichloromethane solution with Mukaiyama's reagent (2-chloro-1-methylpyridinium iodide) to form highly fluorescent ester derivatives that can be separated by silica normal-phase high-performance liquid chromatography (HPLC). Rhodamine WT and Oregon green 488 were also useful derivatization reagents. The detection limit for detection of cholesterol and bryostatin by Bodipy FL was less than 1fmol. The reaction conditions are gentle enough that low concentrations of unstable alcohols such as bryostatin 1 can be measured.
authors.affiliation: Blanchette Rockefeller Neurosciences Institute, Morgantown, WV 26505, USA. tjnelson@brni-jhu.org
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Nelson
mesh_terms: D000438:Alcohols / Q000737:chemistry*; D000873:Anthracenes / Q000737:chemistry; D054713:Bryostatins / Q000737:chemistry; D002175:Candida / Q000737:chemistry; D002264:Carboxylic Acids / Q000737:chemistry; D002784:Cholesterol / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003374:Coumarins / Q000737:chemistry; D004801:Eosine Yellowish-(YS) / Q000737:chemistry; D004951:Esterification; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005453:Fluorescence; D006834:Hydrazines / Q000737:chemistry; D007202:Indicators and Reagents / Q000737:chemistry; D008049:Lipase / Q000737:chemistry; D011726:Pyridinium Compounds / Q000737:chemistry; D012235:Rhodamines / Q000737:chemistry; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000438:Alcohols; D000873:Anthracenes; D054713:Bryostatins; D002264:Carboxylic Acids; D003374:Coumarins; D006834:Hydrazines; D007202:Indicators and Reagents; C436864:Oregon Green 488 carboxylic acid; D011726:Pyridinium Compounds; D012235:Rhodamines; C075233:7-diethylaminocoumarin-3-carbohydrazide; C222011:rhodamine WT; C048173:2-chloro-1-methylpyridinium; C061472:9-anthranoylnitrile; D002784:Cholesterol; D008049:Lipase; D016650:Fluorescein-5-isothiocyanate; D004801:Eosine Yellowish-(YS)