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

Springer Science and Business Media LLC - Tập 67 - Trang 237-243 - 2008
Uttam Mandal1, Amlan Kanti Sarkar1, Kadagi Veeran Gowda1, Sangita Agarwal1, Anirbandeep Bose1, Uttam Bhaumik1, Debotri Ghosh1, Tapan Kumar Pal1
1Bioequivalence Study Centre, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India

Tóm tắt

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.

Tài liệu tham khảo

Tassone DM, Boyce E, Guyer J, Nuzum D (2007) Clin Ther 29:26-48 Hamandi K, Sander JW (2006) Seizure 15:73-78 Kavoussi R (2006) Eur Neuropsychopharmacol 16:S128-S133 Freynhagen R, Strojek K, Griesing T, Whalen E, Balkenohl M (2005) Pain 115:254-263 Sharma U, Young J, LaMoreaux L (2005) Pain 6:S29 Arroyo S, Anhut H, Kugler AR, Lee CM, Knapp LE, Garofalo EA, Messmer S (2004) Epilepsia 45:20-27 Uthman BM, Lee CM, Spiegel K (2004) Epilepsia 45:1-393 Lydiard RB, Feltner DE, Robinson P, Liu-Dumaw M, Pande AC (2001) Eur Neuropsychopharmacol 11:S159-S160 Lauria HBA, Pohl RB (2003) Expert Opin Invest Drugs 12:663-672 Dooley DJ, Donovan CM, Meder WP, Whetzel SZ (2002) Synapse 45:171-190 Gee NS, Brown JP, Dissanayake VUK, Offord J, Thurlow R, Woodruff GN (1996) J Biol Chem 271:5768-5776 Fink K, Dooley DJ, Meder WP, Chauhan NS, Duffy S, Clusmann H, Gothert M (2002) Neuropharmacology 42:229-236 Dooley DJ, Donovan CM, Pugsley TA (2000) J Pharmacol Exp Ther 295:1086-1093 Dooley D J, Mieske CA, Borosky SA (2000) Neurosci Lett 280:107-110 Maneuf YP, Hughes J, McKnight AT (2001) Pain 93:191-196 Bialer M, Johannessen SI, Kupferberg HJ, Levy RH, Perucca E, Tomson T (2007) Epilepsy Res 73:1-52 Taylor CP, Angelotti T, Fauman E (2007) Epilepsy Res 73:137-150 Wesche D, Bockbrader H (2005) Pain 6:29 Stewart BH, Kugler AR, Thompson PR, Bockbrader HN (1993) Pharm Res 10:276-228 Turck D, Vollmer KO, Bockbrader H (1989) Eur J Clin Pharmacol 36:310 Vermeij TAC, Edelbroek PM (2004) J Chromatogr B 810:297-303 Berry D, Millington C (2005) Ther Drug Monit 27:451-456 Food and Drug Administration of the United States (2001) Guidance for industry-Bioanalytical Method Validation, U.S. Department of Health and Human Services, Center for Drug Evaluation and Research (CDER), Center for Veterinary Medicine (CVM). http://www.fda.gov/cder/guidance/index.htm