Bioanalytical method development and validation of rizatriptan in human plasma using LC–MS/MS method

Aim The aim of the present work was to develop a novel, reliable and accurate Liquid Chromatography–Mass Spectrometry/Mass spectrometry (LC–MS/MS) method for the estimation of a potent 5-HT1B/1D receptor agonist, Rizatriptan in human plasma and to validate the proposed method. Experimental Zolmitriptan was used as the internal standard. The analyte and internal standard were isolated from 100 mL plasma samples by liquid-liquid extraction (LLE) and chromatographed on an ACE C18 column, (150 × 4.6 mm, 3 μm) with a mobile phase consisting of acetonitrilee10 mM aqueous ammonium acetate-acetic acid (90:10:0.5% v/v/v) pumped at 1.0 mL/min. The method had a chromatographic total run time of 10 min. A Varian 1200 L electrospray tandem mass spectrometer equipped with an electrospray ionization source was operated in multiple reaction monitoring (MRM) mode with the precursor-to-product ion transitions m/z 270.39/201.3 (Rizatriptan) and 288.43/182.2 (Zolmitriptan) used for quantitation. Results and discussion The method was validated over the concentration range of 50–1000 ng/ml and was found to have acceptable accuracy, precision, linearity, and selectivity. The mean extraction recovery from spiked plasma samples was above 98.26–101.32%. The intra-day accuracy of the assay ranged from 99.525 to 103.32% and intra-day precision ranged from 99.525 to 103.32% C.V. Inter-day accuracy and precision results for quality control samples ranged between 103.62 and 105.196% of nominal and precision is observed to be 5.45–10.946% C.V. The drug was found to be stable after a number of stability studies. Conclusion According to the validated results, the proposed method was found to be specific, accurate, and precise and could be used for the estimation of Rizatriptan in human plasma over a concentration range of 50.0–1000 ng/ml and can be applied for the routine analysis.