Development and Validation of a Highly Sensitive LC-MS/MS Method for the Quantification of Gabapentin in Human Plasma: Analytical Method Validation and Application in Bioequivalence Studies

In this study, a novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the quantification of gabapentin in human plasma. The method employed a reverse-phase high-performance liquid chromatography (RP-HPLC) system coupled with an electrospray ionization (ESI) quadrupole mass spectrometer. Analytical method validation (AMV) was performed in compliance with the International Council for Harmonisation (ICH) M۱۰ guidelines and European Medicines Evaluation Agency (EMEA) recommendations. Gabapentin and the internal standard (tramadol) were extracted from plasma samples using a liquid-liquid extraction technique, followed by chromatographic separation on an Agilent ZORBAX SB-C۱۸ column. The method demonstrated excellent specificity, linearity, and precision over a concentration range of ۰.۲ to ۹۶۰۰ ng/mL. The limit of quantification (LLOQ) for gabapentin was determined to be ۲۰ ng/mL with a signal-to-noise ratio (S/N) greater than ۱۰. The accuracy of the method was assessed by testing at three different concentration levels (low, medium, and high), with deviation percentages (Dev%) of ≤ ۵.۰%. The method exhibited satisfactory precision, with relative standard deviations (RSD%) ≤ ۱۰%. Cross-validation between between-run and within-run conditions confirmed method robustness and reliability. The method's applicability was demonstrated by its successful use in plasma bioequivalence studies involving gabapentin. Additionally, the carry-over effect was tested to ensure minimal interference in subsequent injections. Overall, the method is highly suitable for pharmacokinetic studies and clinical research requiring precise quantification of gabapentin in human plasma.

کلیدواژه ها:

Gabapentin ، LC-MS/MS ، Plasma quantification ، Method validation ، ICH M۱۰ ، EMEA guidelines ، Bioequivalence ، Analytical method validation ، Electrospray ionization ، Liquid-liquid extraction