Analytical quality by design approach for the control of potentially counterfeit chloroquine with some NSAIDS using HPLC with fluorescence detection in pharmaceutical preparation and breast milk

Abstract Chloroquine phosphate (CQ) the antimalarial drug and suggested to treat pandemic disease coronavirus (COVID-19) is often adulterated with some of non-steroidal anti-inflammatory drugs (NSAIDs) such as paracetamol, aspirin (ASP), or both. The purpose this study detect counterfeited drugs, using a reversed phase high pressure liquid chromatography (RP-HPLC) method fluorescence detection. Analysis was divided into three phases. In first phase, Plackett-Burman design (PBD) used screen five independent factors, namely, buffer pH, concentration (mM), acetonitrile content (%), flow rate (mL/min) triethylamine (TEA) in preparation (%). selected dependent variables were (resolution, symmetry peaks run time). objective second optimize performance Box-Behnken (BBD) desirability function for multiple response optimization obtain best chromatographic shortest time. Optimal separation achieved on YMC-pack pro C18 ODS-A column (15 cm × 4.6 mm, 5 µm) at room temperature optimum mobile consisted mM sodium dihydrogen containing 0.5% triethyamine (30:70, v/v) pH adjusted 3.5 an orthophosphoric acid solution. maintained 1 mL/min, detection performed detector fixed 380 nm (λ emission ) after excitation 335 ). third validation according ICH guidelines, providing be specific, precise, accurate, robust. linear over range 0.4–8 µg/mL chloroquine ASP, while paracetamol it 16–48 µg/mL. developed RP-HPLC quantitation dosage form samples. shows great tendency classification between genuine ones pharmaceutical preparations breast milk.