High Sensitive Method Development and validation of lmotriptan in Human Plasma by UPLC Tandem Mass Spectrometry

A highly sensitive ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed for the quantification of Almotriptan in human plasma.To the best of our knowledge; this is the sensitive method for the quantification of Almotriptan in human plasma. Matrix effects were strongly reduced and selectivity increased by using an efficient chromatographic separation on a sub-2m column (Acquity UPLC HSS T3 1.8 m, 2.1 50 mm) in 3 min with a gradient of ammonium formate 5mM and acetonitrile as mobile phase at a flow rate of 0.4 ml/min. Detection was made using a tandem quadrupole mass spectrometer operating in positive electrospray ionization mode, using multiple reaction monitoring. The procedure was fully validated according to the latest Food and Drug Administration guidelines. Very good results were obtained by using Naratriptan as internal standard, to compensate for the variability due to the extraction and ionization steps. The method was very sensitive with lower limits of quantification of 0.1 ng/ml for Almotriptan. The upper limit of quantification was 250 ng/ml. Accuracy (94.0-105.7%), Precision (1.0-6.7%), and internal standard-normalized matrix effects (95-102%) were in accordance with international recommendations. The method can be used for pharmacokinetic studies.