title: Development and validation of an LC-MS-MS method for the simultaneous determination of sulforaphane and its metabolites in rat plasma and its application in pharmacokinetic studies.
abstract: A highly sensitive and simple high-performance liquid chromatographic-tandem mass spectrometric (LC-MS-MS) assay is developed and validated for the quantification of sulforaphane and its metabolites in rat plasma. Sulforaphane (SFN) and its metabolites, sulforaphane glutathione (SFN-GSH) and sulforaphane N-acetyl cysteine (SFN-NAC) conjugates, are extracted from rat plasma by methanol-formic acid (100:0.1, v/v) and analyzed using a reversed-phase gradient elution on a Develosil 3 μm RP-Aqueous C(30) 140Å column. A 15-min linear gradient with acetonitrile-water (5:95, v/v), containing 10 mM ammonium acetate and 0.2% formic acid, as mobile phase A, and acetonitrile-water (95:5, v/v), containing 10 mM ammonium acetate and 0.2% formic acid as mobile phase B, is used. Sulforaphane and its metabolites are well separated. Sulforaphene is used as the internal standard. The lower limits of quantification are 1 ng/mL for SFN and 10 ng/mL for both SFN-NAC and SFN-GSH. The calibration curves are linear over the concentration range of 25-20,000 ng/mL of plasma for each analyte. This novel LC-MS-MS method shows satisfactory accuracy and precision and is sufficiently sensitive for the performance of pharmacokinetic studies in rats.
authors.affiliation: Graduate Program in Pharmaceutical Sciences, Ernest-Mario School of Pharmacy, Rutgers, The State University of New Jersey, Piscataway, 08854, USA.
authors.forename: Hu
authors.initials: H
authors.lastname: Wang
authors.forename: Wen
authors.initials: W
authors.lastname: Lin
authors.forename: Guoxiang
authors.initials: G
authors.lastname: Shen
authors.forename: Tin-Oo
authors.initials: TO
authors.lastname: Khor
authors.forename: Amin A
authors.initials: AA
authors.lastname: Nomeir
authors.forename: Ah-Ng
authors.initials: AN
authors.lastname: Kong
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D017879:Isothiocyanates / Q000097:blood; D016018:Least-Squares Analysis; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013454:Sulfoxides; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013861:Thiocyanates / Q000097:blood* / Q000493:pharmacokinetics
chemical_list: D017879:Isothiocyanates; D013454:Sulfoxides; D013861:Thiocyanates; C016766:sulforaphane; C473643:sulphoraphene