Topkafa, MustafaAyyildiz, Hamide FilizMemon, Fakhar N.Kara, Huseyin2020-03-262020-03-2620161615-93061615-9314https://dx.doi.org/10.1002/jssc.201600045https://hdl.handle.net/20.500.12395/33825A new humic acid based stationary phase has been used, for the first time, to achieve the separation and quantification of paracetamol and caffeine in pharmaceutical preparations under reversed-phase high-performance liquid chromatography conditions. Central composite design was applied as a powerful tool to optimize the most dominant parameters that influence the resolution of reversed-phase high-performance liquid chromatography, that is, mobile phase composition (acetonitrile percentage in water), flow rate, and column temperature. The optimum conditions were obtained as 21.69%, 1.5 mL/min, and 15 degrees C, respectively, with the aid of a second-order quadratic model and desirability function. Under the optimum conditions, the peaks could be baseline separated within 10 min. For the developed reversed-phase high-performance liquid chromatography method, the linearity was investigated in the concentration ranges of 2-160 mg/mL (R-2 = 0.999) for paracetamol and 2-9.9 mg/mL (R-2 = 0.991) for caffeine. Mean recoveries for paracetamol and caffeine were 95.90 and 95.68%, respectively. The limits of detection and quantification were 4.1 x 10(-4) and 1.3 x 10(-3) mg/mL for paracetamol and 1.6 x 10(-4) and 5.0 x 10(-4) mg/mL for caffeine. The results showed that the new humic acid based stationary phase is very suitable for the separation of paracetamol and caffeine in pharmaceutical preparations and, thus it can be used effectively in the pharmaceutical industry.en10.1002/jssc.201600045info:eu-repo/semantics/closedAccessCaffeineCentral composite designImmobilized humic acidParacetamolReversed-phase high-performance liquid chromatographyArticle39132451245827197723Q2WOS:000379905600002Q2