Feasible salting-out assisted extraction integrated with green TLC approach for concurrent chlorzoxazone and ibuprofen determination in human plasma.

The combination of chlorzoxazone and ibuprofen provides an effective therapeutic option for relieving pain and inflammation associated with rheumatoid arthritis, sprains, ankylosing spondylitis, and pain after surgical procedures. Given its widespread use globally, it is crucial to develop a straightforward method for determination of this combination in real human plasma. This study focuses on the salting-out liquid-liquid extraction technique, an emerging approach for extracting drugs from biological matrices. A salting out liquid-liquid extraction with ammonium sulfate salt and acetonitrile has been utilized to extract chlorzoxazone and ibuprofen from real human plasma prior to a TLC-densitometric approach for their separation and quantitative determination. As a stationary phase, TLC silica gel 60 F254 plates have been employed, while ethyl acetate: methanol: ammonia (8:2:0.5 v/v/v) was used as a mobile phase. Both drugs have been detected with dual wavelengths at 223 and 282 nm for ibuprofen and chlorzoxazone, respectively. Separate and distinct bands have been detected with retardation factors of 0.34 and 0.76 for ibuprofen and chlorzoxazone, respectively. The suggested approach revealed quantitation limits of 5 and 19 µg. mL-1 and detection limits of 2 and 6 µg. mL-1 over a linear range of 5-200 and 20-200 µg. mL-1 for chlorzoxazone and ibuprofen, respectively. In line with the growing emphasis on sustainable practices, the greenness of the proposed method was evaluated using the Analytical Eco-Scale and the Green Analytical Procedure Index (GAPI) tools.