title: Reliable and sensitive determination of dutasteride in human plasma by liquid chromatography-tandem mass spectrometry.
abstract: An accurate and precise method was developed and validated using LC-MS/MS to quantify dutasteride in human plasma. The analyte and dutasteride-13C6 as internal standard (IS) were extracted from 300 μL plasma volume using methyl tert-butyl ether-n-hexane (80:20, v/v). Chromatographic analysis was performed on a Gemini C18 (150 × 4.6 mm, 5 µm) column using acetonitrile-5 mm ammonium formate, pH adjusted to 4.0 with formic acid (85:15, v/v) as the mobile phase. Tandem mass spectrometry in positive ionization mode was used to quantify dutasteride by multiple reaction monitoring. The entire data processing was done using Watson LIMS(TM) software, which provided excellent data integrity and high throughput with improved operational efficiency. The calibration curve was linear in the range of 0.1-25 ng/mL, with intra-and inter-batch values for accuracy and precision (coefficient of variation) ranging from 95.8 to 104.0 and from 0.7 to 5.3%, respectively. The mean overall recovery across quality controls was ≥95% for the analyte and IS, while the interference of matrix expressed as IS-normalized matrix factors ranged from 1.01 to 1.02. The method was successfully applied to support a bioequivalence study of 0.5 mg dutasteride capsules in 24 healthy subjects. Assay reproducibility was demonstrated by reanalysis of 103 incurred samples.
authors: Bioanalytical Department, Veeda Clinical Research, Ahmedabad, 387810, India.
authors: Pritesh
authors: P
authors: Contractor
authors: Hemal
authors: H
authors: Kurani
authors: Swati
authors: S
authors: Guttikar
authors: Pranav S
authors: PS
authors: Shrivastav
mesh_terms: D000328:Adult; D001378:Azasteroids / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D000068538:Dutasteride; D006801:Humans; D016014:Linear Models; D059625:Liquid-Liquid Extraction / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013810:Therapeutic Equivalency
chemical_list: D001378:Azasteroids; D000068538:Dutasteride
keywords: LC-MS/MS; bioequivalence study; dutasteride; dutasteride-13C6; human plasma; liquid-liquid extraction