Bioanalytical method development and validation of trigonelline by tandem mass spectra and its application to pharmacokinetic study

Abstract

A rapid, simple and sensitive LC–MS/MS analytical method was developed and validated for the determination of trigonelline in Sprague Dawley Rat Plasma, using Atenolol as an internal standard. AB SCIEX QTRAP® 4000 LC-MS/MS was used. Chromatographic separation was achieved using a Gemini 5µ C6 Phenyl 100x4.6mm (Phenomenex), maintained at 40°C. The samples were eluted using 0.1% Formic acid in Water (solvent A) and 0.1% formic acid in Acetonitrile (solvent B), at a flow rate of 0.8 ml/min with a total run time of 4.0 min. The atmospheric pressure ionization source (API-4000) triple quadruple mass spectrometer equipped with an electro spray ionization source, operating in positive mode. Analysis was performed in multiple reaction-monitoring (MRM) mode by monitoring the ion transitions from m/z 138.0→92.1 (Trigonelline) and m/z 267.3→145.2 (IS). Calibration curves in spiked plasma were linear over the concentration range of 1–2000 ng/mL with determination coefficient >0.9989. The developed method was validated in terms of selectivity, accuracy, precision, linearity, Matrix effect, dilution integrity and stability study. The proposed method uses less biological material and the method is compatible for different biological matrix also. Method can be applicable for pharmacokinetic studies and in-vitro related studies using LC-MS/MS or HPLC.