Evaluation of Matrix Effect in Bio Estimation of Prasugrel Hydrochloride by Liquid Chromatography-Mass Spectrometry

Liquid chromatography-mass spectrometry (LC-MS) is sensitive technique that is affected by matrix effect. It arises due to phospholipids, proteins or metabolites present in biological fluids. It either increases or decreases the ionization of sample and affects detection limits. By careful evaluation of matrix effect on the drugs and their ionization in LC-MS the sensitivity could be improved. Evaluation of matrix effect for bio estimation of prasugrel hydrochloride is done by sample preparation methods, chromatographic conditions, internal standard and mass spectrometric conditions. A LC-MS method was developed for prasugrel hydrochloride with mobile phase consisted of ammonium formate (5mM, pH 7.5): acetonitrile (30:70, %v/v) at 1 mL/min flow rate and PDA detection at 220 nm. The mass detection method was after ionization with ESI probe, CDL temperature 230°C, detector voltage-1.5 Kv, nebulisation gas flow-1.5 L/min and column temperature was 30.5°C. The molecular ion peak was identified at m/z of 410 and product ion peak at m/z 374. The plasma spiked with prasugrel was subjected to each evaluation process and matrix effect was studied.  In sample preparation technique, the extraction efficiency and process efficiency were also calculated. The study results have shown that the matrix effect could be reduced systematically by the procedures performed and the sensitivity of LC-MS in the detection and quantification was increased to detect 20 ng/mL of Prasugrel from plasma.