Analysis of Aspirin, Prasugrel and Clopidogrel in Counterfeit Pharmaceutical and Herbal Products: Plackett–Burman Screening and Box–Behnken Optimization

Abstract

An isocratic reversed-phase high-performance liquid chromatographic method has been developed and validated for the simultaneous determination of aspirin, prasugrel HCl and clopidogrel bisulfate in the presence of clopidogrel-related compound (impurity-A) in focus on counterfeit. This method was used to determine counterfeited antiplatelet drugs in two substandard Indian pharmaceutical products sold on the market in Yemen and two traditional herbal medicines sold on the market in China. Thin layer chromatography and mass spectrometry of counterfeit herbal medicines have additionally been carried out to verify the identification of adulterants. Chromatographic separation was performed on Inertsil ® ODS-3 C18 (4.6 × 250 mm, 5 μm) with isocratic mobile phase elution containing a mixture of acetonitrile: (25 mM) potassium dihydrogen phosphate buffer, pH 2.7 adjusted with 0.1 M o-phosphoric acid (79: 21, v/v), at a flow rate of 1 mL/min and UV detection at 220 nm. Designs of experiment methodology, Plackett–Burman and Box–Behnken designs were used for the screening and optimization of the mobile phase composition. The method validation was also performed in accordance with the International Council on Harmonization (ICH) guidelines. The method developed for routine analysis was found to be sensitive, simple, accurate and highly robust. The results were statistically compared to reference methods using Student’s t-test and variance ratio F-test at P < 0.05.

Utilizing experimental design and desirability function in optimizing RP-HPLC method for simultaneous determination of some skeletal muscle relaxants and analgesics

“…The Adjusted R-squared value serves as a more reliable measure of the model's goodness of fit than the R-squared value, especially in situations where the model incorporates an excessive number of independent variables. High values of R-squared and adjusted R-squared indicate that the data have been fitted properly by the models 39 . As noted in Table 4 , the predicted R-squared values were within accepted limits with regards to the adjusted R-squared values, i.e.…”

Development and Validation of RP-HPLC Method for Analysis of Counterfeit Clopidogrel

“…Different volumes (0.5, 1.5, 2, 2.5, and 3.5) mL of clopidogrel bisulfate and (0.3, 0.5, 1.5, 2, and 2.5) mL of acetyl salicylic acid were taken from the standard stock solution of each drug into 10 mL volumetric flasks and the volume was completed to the mark with methanol to obtain different concentrations (10,30,40, 50 and 70) µg/mL and (6, 10, 30, 40 and 50) µg/mL for clopidogrel bisulfate and acetyl salicylic acid, respectively to construct the calibration curves.…”

“…Counterfeit clopidogrel bisulfate was analyzed by different methods such as RP-HPLC, TLC, and mass spectroscopy 3,10 . Few analytical methods were reported for the simultaneous determination of clopidogrel and acetyl salicylic acid by RP-HPLC 11 and chemometric techniques 12 .…”