Abstract
A bioanalytical method has been developed and validated for determination of pregabalin in human plasma. The analytical method consists in the precipitation of plasma sample with trichloro acetic acid (20% v/v solution in water), followed by the determination of pregabalin by an LC-MS-MS method using gabapentin as internal standard. Separation was achieved on a Gemini C18 50 mm × 2.0 mm (3 μm) column with an isocratic mobile phase consisting of methanol–water (98:2, v/v) with 0.5% v/v formic acid. Protonated ions formed by a turbo ionspray in positive mode were used to detect analyte and internal standard. The MS-MS detection was by monitoring the fragmentation of 160.2→55.1 (m/z) for pregabalin and 172.2→67.1 (m/z) for gabapentin on a triple quadrupole mass spectrometer. The assay was calibrated over the range 0.1–15.0 μg mL−1 with correlation coefficient of 0.9998. Validation data showed intra-batch (n = 6) CV% ≤ 6.89 and RE (%) between −4.17 and +3.08 and inter-batch (n = 18) CV% < 9.09 and RE (%) between −3.0 and +10.00. Mean extraction recovery were 80.45–89.12% for three QC samples and 87.56% for IS. Plasma samples were stable for three freeze–thaw cycles, or 24 h ambient storage, or 1 and 3 months storage at −20 °C. Processed sample (ready for injection) were stable up to 72 h at autosampler (4 °C). This method has been used for analyzing plasma samples from a bioequivalence study with 18 volunteers.
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Acknowledgments
The authors are thankful to Burgeon Pharmaceutical Pvt. Ltd., Chennai, India, and Cosmas Pharmaceuticals, Ludhiana, India for supplying the gift samples of pregabalin and gabapentin respectively. The authors also acknowledge the Department of Science and Technology (DST), Govt. of India, New Delhi, for providing financial assistance to carry out this project through their project No. VII-PRDSF/56/05-06 under DPRP programme.
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Mandal, U., Sarkar, A.K., Gowda, K.V. et al. Determination of Pregabalin in Human Plasma Using LC-MS-MS. Chroma 67, 237–243 (2008). https://doi.org/10.1365/s10337-007-0440-2
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DOI: https://doi.org/10.1365/s10337-007-0440-2