title: Application of a UPLC-MS/MS method for the analysis of alosetron in human plasma to support a bioequivalence study in healthy males and females.
abstract: A simple, rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method has been developed and validated for the determination of alosetron (ALO) in human plasma. The assay method involved solid-phase extraction of ALO and ALO 13C-d3 as internal standard (IS) on a LichroSep DVB-HL (30 mg, 1 cm(3) ) cartridge. The chromatography was performed on an Acquity UPLC BEH C18 (50 × 2.1 mm, 1.7 µm) column using acetonitrile and 2.0 mm ammonium formate, pH 3.0 adjusted with 0.1% formic acid (80:20, v/v) as the mobile phase in an isocratic mode. For quantitative analysis, the multiple reaction monitoring transitions studied were m/z 295.1/201.0 for ALO and m/z 299.1/205.1 for IS in the positive ionization mode. The method was validated over a concentration range of 0.01-10.0 ng/mL for ALO. Post-column infusion experiment showed no positive or negative peaks in the elution range of the analyte and IS after injection of extracted blank plasma. The extent of ion-suppression/enhancement, expressed as IS-normalized matrix factor, varied from 0.96 to 1.04. The assay recovery was within 97-103% for ALO and IS. The method was successfully applied to support a bioequivalence study of 1.0 mg alosetron tablets in 28 healthy Indian male and female subjects.
authors.affiliation: Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad, 380009, Gujarat, India.
authors.forename: Darshan V
authors.initials: DV
authors.lastname: Chaudhary
authors.affiliation: Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad, 380009, Gujarat, India.
authors.forename: Daxesh P
authors.initials: DP
authors.lastname: Patel
authors.affiliation: Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad, 380009, Gujarat, India.
authors.forename: Jaivik V
authors.initials: JV
authors.lastname: Shah
authors.affiliation: Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad, 380009, Gujarat, India.
authors.forename: Priyanka A
authors.initials: PA
authors.lastname: Shah
authors.affiliation: Department of Chemistry, St Xavier's College, Navrangpura, Ahmedabad, 380009, Gujarat, India.
authors.forename: Mallika
authors.initials: M
authors.lastname: Sanyal
authors.affiliation: Department of Chemistry, School of Sciences, Gujarat University, Ahmedabad, 380009, Gujarat, India.
authors.forename: Pranav S
authors.initials: PS
authors.lastname: Shrivastav
mesh_terms: D000293:Adolescent; D000328:Adult; D002138:Calibration; D002243:Carbolines / Q000097:blood* / Q000493:pharmacokinetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018592:Cross-Over Studies; D005260:Female; D064368:Healthy Volunteers; D006801:Humans; D008297:Male; D008875:Middle Aged; D015203:Reproducibility of Results; D052616:Solid Phase Extraction / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013810:Therapeutic Equivalency; D055815:Young Adult
chemical_list: D002243:Carbolines; C090840:alosetron
keywords: Alosetron; UPLC-MS/MS; gender effects on pharmacokinetics; high throughput; solid phase extraction