title: Quantitative liquid chromatographic determination of sanguinarine in cell culture medium and in rat urine and plasma.
abstract: Sanguinarine is a quaternary benzo[c]phenanthridine alkaloid, extracted from the argemone oil, which produced severe human intoxications. To investigate the sanguinarine biotransformation, we develop a simple extraction process and a high performance liquid chromatographic separation coupled to a sensitive fluorometric detection of sanguinarine in cell culture medium, as well as in rat urine and plasma. After extraction with an acidified organic solvent, sanguinarine elution is performed within 15 min on a Nucleosil C18 column with a gradient using 0.2% formic acid/water/acetonitrile as mobile phase. Extracted and standard sanguinarine are characterized by mass spectrometry. The extraction recovery of sanguinarine is about 80% in cell culture medium and in rat urine, but lower in plasma. This convenient high performance liquid chromatography (HPLC) method allows to quantify sanguinarine over concentrations ranged 10-2000 ng ml(-1). The limit of fluorometric detection is 0.5 ng. Under these conditions, the lower limit of quantification of sanguinarine is 50 ng ml(-1) in cell culture medium and in rat urine and 100 ng ml(-1) in rat plasma. This analytical HPLC method is specific, linear and reproducible in all media and is suitable for quantitative determination of sanguinarine in biological fluids.
authors.affiliation: CNRS UPR 2301, Institut de Chimie des Substances Naturelles, Avenue de la Terrasse, 91198 Cedex, Gif sur Yvette, France. hoelling@icsn.cnrs-gif.fr
authors.forename: Henri
authors.initials: H
authors.lastname: Hoellinger
authors.forename: Micheline
authors.initials: M
authors.lastname: Re
authors.forename: Alain
authors.initials: A
authors.lastname: Deroussent
authors.forename: Ravindra Pratap
authors.initials: RP
authors.lastname: Singh
authors.forename: Thierry
authors.initials: T
authors.lastname: Cresteil
mesh_terms: D000470:Alkaloids / Q000097:blood* / Q000652:urine*; D000818:Animals; D053119:Benzophenanthridines; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003470:Culture Media / Q000737:chemistry*; D007546:Isoquinolines; D008297:Male; D013058:Mass Spectrometry; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D000470:Alkaloids; D053119:Benzophenanthridines; D003470:Culture Media; D007546:Isoquinolines; C005705:sanguinarine