Rapid quantification of nebivolol in human plasma by liquid chromatography coupled with electrospray ionization tandem mass spectrometry.
A simple, sensitive and rapid liquid chromatographic/electrospray ionization tandem mass spectrometric method was developed and validated for the quantitation of nebivolol in human plasma. The method involved a simple single-step liquid-liquid extraction with diethyl ether/dichloromethane (70/30). The analyte was chromatographed on Waters symmetry C18 reversed-phase chromatographic column by isocratic elution with water:acetonitrile:formic acid (30:70:0.03, v/v) and analyzed by mass spectrometry in the multiple reaction monitoring mode. The precursor to product ion transitions of m/z 406.4-151.5 and m/z 409.1-228.1 were used to measure the analyte and the internal standard (I.S.), respectively. The chromatographic runtime was 2 min and the weighted (1/x2) calibration curves were linear over the range 50-10,000 pg/mL. The method was validated in terms of accuracy, precision, absolute recovery, freeze-thaw stability, bench-top stability and re-injection reproducibility. The limit of detection and lower limit of quantification in human plasma were 10 and 50 pg/mL, respectively. The within- and between-batch accuracy and precision were found to be well within acceptable limits (<10%). The analyte was stable after three freeze-thaw cycles (deviation <10%). The average absolute recoveries of nebivolol and tamsulosin, used as an internal standard, from spiked plasma samples were 73.4+/-3.7 and 72.1+/-2.0%, respectively. The assay method described here was applied to study the pharmacokinetics of nebivolol.
Ramakrishna
N V S
NV
Biopharmaceutical Research, Suven Life Sciences Ltd., Serene Chambers, Road #7, Banjara Hills, Hyderabad 500034, India. nvsrk@suven.com
Vishwottam
K N
KN
Koteshwara
M
M
Manoj
S
S
Santosh
M
M
Varma
D P
DP
D000319:Adrenergic beta-Antagonists / Q000097:blood* / Q000493:pharmacokinetics; D001578:Benzopyrans / Q000097:blood* / Q000493:pharmacokinetics; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004983:Ethanolamines / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D000068577:Nebivolol; D011786:Quality Control; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization
D000319:Adrenergic beta-Antagonists; D001578:Benzopyrans; D004983:Ethanolamines; D000068577:Nebivolol