Interest in implementing green chemistry principles in analytical chemistry has grown dramatically in the past few years. The solvents used have the major influence on the greenness of the method. Most conventional high-performance liquid chromatography methods employed utilize solvents that are "hazardous for the environment". In the present study, two-factor three-level response surface design was exploited to develop eco-friendly chromatographic methods for two different mixtures. The first one was atorvastatin and amlodipine and the second one was amlodipine, perindopril, and indapamide. As it is nontoxic to the environment, ethanol was used as the organic modifier in the mobile phase. The separation of the first mixture was attained using phosphate buffer (pH 7)/ethanol (42:58 v/v), and the second mixture was fully resolved using phosphate buffer (pH 5)/ethanol (40:60 v/v). The use of high-performance liquid chromatography allows excellent resolution in a short run time, hence, less waste was produced. The greenness of the developed methods was assessed by two evaluation tools, namely, National Environmental Methods Index and analytical eco-scale, and found to be excellent green analytical methods. Moreover, the developed methods were compared with other reported methods regarding accuracy and greenness and were found to be perfect alternatives to reported methods for separation and quantification of the mixtures.

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Determination of a widely used antihypertensive combination of atenolol and hydrochlorothiazide was achieved by rapid and eco-friendly high-performance liquid chromatography method combined with fluorescence detection. The response surface methodology is conducive to the complete separation of the two drugs in a shorter analysis time. The separation of the mixture was achieved using an Inertsil C18 analytical column (150 × 4.6 mm, 5 µ). The mobile phase used was ethanol: potassium dihydrogen phosphate at pH 3 (65:35 v/v) and the flow rate was 0.7 mL/min. The fluorescence detector operated at excitation and emission wavelengths of 230 and 310 nm (atenolol) and 270 and 375 nm (hydrochlorothiazide). The linearity of the developed method covered a concentration of atenolol of 0.05-5 µg/mL and a concentration of hydrochlorothiazide of 0.02-1 µg/mL. The greenness of the developed method was evaluated by analytical eco-scale and the recently reported evaluation method, that is, green analytical procedure index, and it was found to be an excellent, sensitive, and green alternative to the reported methods. The developed method was validated according to the ICH guidelines and compared with the reference method. No significant difference was found in terms of accuracy.

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