Determination of Pregabalin in Human Plasma Using LC-MS-MS

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 x 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[rightward arrow]55.1 (m/z) for pregabalin and 172.2[rightward arrow]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%