BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF PREGABALIN IN RAT PLASMA BY SOLID PHASE EXTRACTION WITH HPLC-MS/MS: APPLICATION TO A PHARMACOKINETIC STUDY

Abstract

The most suitable bioanalytical method based on solid phase extraction has been developed and validated for quantification of pregabalin in rat plasma. Pregabalin D4 was used as an internal standard for pregabalin. Thermo Hypurity C18 (4.6 mm × 150 mm, 5.0 µm) column provided chromatographic separation of analyte followed by detection with mass spectrometry. The method involved simple isocratic chromatographic separation and mass spectrometric detection in the positive ionization mode using an API-4000 system. The total run time was 3.5 minutes. The proposed method has been validated with the linear range of 0.50–20000.00 ng/mL for Pregabalin. The intra-run and inter-run precision values were within 1.05 to 4.81% and 1.57 to 3.90%, respectively. The overall recovery for pregabalin and pregabalin D4 was 86.49% and 86.57%, respectively. This validated method was successfully applied into the pharmacokinetic study of rat plasma.

Keywords:

• anticonvulsant
• HPLC-MS/MS
• pharmacokinetic study
• pregabalin
• rat plasma
• solid-phase extraction

ACKNOWLEDGMENT

The authors are grateful to the Indian Institute of Chemical Technology, Hyderabad, for literature survey and AZIDUS, Chennai, India for their Lab facility of this research work.

Notes

AUC_0—∞: area under the curve extrapolated to infinity; AUC_0—t: area under the curve up to the last sampling time; Cmax: the maximum plasma concentration; Tmax: the time to reach peak concentration; Kel: the apparent elimination rate constant.