title: Quantitative determination of nebivolol from human plasma using liquid chromatography-tandem mass spectrometry.
abstract: In the present work, a rapid, sensitive, specific, precise and accurate liquid chromatography-tandem mass spectrometry method for determination of nebivolol in human plasma was developed and validated with a large calibration curve range (50-5000 pg/mL) which can be used for routine drug analysis and bioequivalence studies. Liquid-liquid extraction method was used to extract the analyte from the human plasma. The separation was achieved using Waters symmetry, C18, 4.6 × 150 mm, 5 μm column with formic acid in water, 0.01%, v/v: Acetonitrile (40:60) as a mobile phase. A flow rate of 0.8 mL/min, no splitting and run time 2.00 min was used for the chromatographic analysis of nebivolol. Sensitivity of this method was found to be 30 pg/mL. The analyte was analyzed by mass spectrometry in the multiple reaction monitoring mode. A Turbo-Ion spray source was interfaced between the HPLC and triple quadrupole mass spectrometer (MDS Sciex API 4000). The precursor-product ion m/z 406.00-151.00 for nebivolol and m/z 410.20-151.00 for nebivolol-D4 were used for quantification of an analyte and its IS. The method was validated in terms of accuracy, precision, selectivity, absolute recovery, freeze-thaw stability, bench-top stability, dry extract stability, short and long term stock solution stability, wet extract stability and re-injection reproducibility. The within- and between-batch accuracy was found to lie within the range of 87.00-100.40% and within- and between-batch precision was obtained within the range 0.33-8.67%. The mean recovery of all three concentration levels for drug was obtained 67.67% where as the mean recovery of IS was 68.74%. The %RSD value at higher concentration and lower concentration in all stability experiments was within 15%. This method is free from ion suppression, ion enhancement and any type of abnormal ionization.
authors.affiliation: Pharmacy Department, Faculty of Technology and Engineering, the MS University of Baroda, Gujarat, India. jatinnandania@gmail.com
authors.forename: Jatin
authors.initials: J
authors.lastname: Nandania
authors.forename: S J
authors.initials: SJ
authors.lastname: Rajput
authors.forename: Pritesh
authors.initials: P
authors.lastname: Contractor
authors.forename: Pragnesh
authors.initials: P
authors.lastname: Vasava
authors.forename: Bhavik
authors.initials: B
authors.lastname: Solanki
authors.forename: Mohsin
authors.initials: M
authors.lastname: Vohra
mesh_terms: D000328:Adult; D001578:Benzopyrans / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D018592:Cross-Over Studies; D004355:Drug Stability; D004983:Ethanolamines / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D005260:Female; D006801:Humans; D016014:Linear Models; D008297:Male; D000068577:Nebivolol; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013810:Therapeutic Equivalency
chemical_list: D001578:Benzopyrans; D004983:Ethanolamines; D000068577:Nebivolol