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Abstracto

Development and Minimizing the Carryover of A Sensitive and High Throughput LC ESI MS/MS Method for the Quantification of Rizatriptan in Human Plasma and Its Application in a Clinical Trial

Pankaj Joshi, Hanumanth Srikanth Cheruvu, Nitin Vig, Abdulla Ansari, Pankaj Singh

A simple, sensitive, and high throughput liquid chromatographic method coupled with the tandem mass spectrometry method has been developed and validated to quantify rizatriptan in human plasma using rizatriptan D 6 as an internal standard. The analyte and internal standard were extracted from plasma via solid-phase extraction and were separated on a C 18 column using methanol: 5mM ammonium acetate buffer: formic acid (80/20/0.1 v/v/v) as a mobile phase. The mass transitions m/z 270.24→201.10 and m/z 276.21→207.20 were used to measure rizatriptan and rizatriptan D 6 respectively. The proposed method validated for a linear dynamic range of 0.151-52.170 ng/mL with a correlation coefficient ≥0.999, where the regression model (1/×2) was best fitted. The carryover of the method in both aqueous and extracted blank samples after the highest concentration level (52.17 ng/mL) was less than 5% compared to the LLOQ concentration level (0.151 ng/mL). The recovery of rizatriptan at LQC, MQC, HQC levels were 79.7, 82.3, and 79.4%, respectively consistent and reproducible. The within and between batches assay precision (% CV) across the three validation batches (LLQC, LQC, MQC, and HQC) were in the range of 1.0-6.5% and 3.0-8.9% respectively. The validated method has been successfully applied to investigate a clinical pharmacokinetic study.

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