Prasugrel

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.

A simple, reliable, rapid, precise, sensitive and validated RP-HPLC method has been developed to determine Acetaminophen (AI) and Prasugrel (PRA) in synthetic mixture form. Chromatographic separation achieved isocratically on Luna C18 column (5 µm, 150mm x 4.60mm) and acetonitrile: 0.05M ammonium acetate buffer (pH 4.5) in the ratio of 75:25 (v/v) as the mobile phase, at a flow rate of 0.6 mL/min. Detection was carried out at 245 nm. Parameters such as linearity, precision, accuracy, recovery, specificity and ruggedness are studied as reported in the ICH guidelines. The retention times for AI and PRA was found to be 2.25 ± 0.5 and 8.72 ± 0.5 min respectively. Linearity for AI and PRA was in the range of 75-375 μg/mL and 10-50 μg/mL respectively.  The mean recoveries obtained for AI and PRA were 99.58 and 99.48% respectively and RSD was less than 2. The correlation coefficients for all components are close to 1. Developed method was found to be accurate, precise, selective and rapid for simultaneous estimation of AI and PRA.

The present research work discusses the two new, simple, accurate, precise and reproducible UV spectrophotometric methods have been developed and validated for the simultaneous determination of Prasugrel (PRASU) and Aspirin (ASP) in their combined dosage form.  Method- I is based on simultaneous equation method using two wavelengths, 254 nm (λmax of PRASU) and 276 nm (λmax of ASP). Method - II Q‐absorption ratio method using two wavelengths, 274.7 nm (Isoabsorptive point) and 254 nm (λmax of PRASU). Methanol was the solvent used in all methods. This method obeyed Beer’s law in the concentration range of 5-60 µg /ml for PRASU and 20-140 μg/ml ASP. All methods were validated statistically and recovery studies were carried out. Hence, the methods herein described can be successfully applied in quality control of combined pharmaceutical dosage form.