HPLC-fluorescence detection for assay of tramadol binary mixtures with ibuprofen or chlorzoxazone in tablets and plasma: Analytical Eco-Scale and GAPI tools for green assessment

Abstract Tramadol, a strong pain killer known for its addictive problems is either co-administrated or co-formulated with other analgesics muscle relaxants. The power of fluorescence detection in HPLC tested to resolve such mixtures plasma matrix reach the required sensitivity simple sample treatment using just protein precipitation. aim this work was develop an eco-friendly and sensitive method fluorimetric analysis Tramadol two binary Ibuprofen (mixture 1) Chlorzoxazone 2) combined dosage forms spiked plasma. Separation done C 18 column mobile phase acetonitrile water (pH 3.5) gradient elution 1 mL/min flow rate. Detection carried out λ excitation/ emission 220 307 nm, respectively. applied detect real samples after invivo application rats, assure that drugs’ metabolites do not affect selectivity assay. Evaluation greenness proposed semi-quantitative Eco‐Scale new Green Analytical Procedure Index which showed can be greener alternative higher both mixtures. (15 min-assay) linear over concentrations 0.1–10 μg/mL 0.1–33 In addition, validated per ICH as well FDA bioanalytical methods’ validation guidelines.