Integrative AQbD, up-to-date greenness, and whiteness tools for evaluation of a sustainable RP-HPLC method used for simultaneous separation of triple antihypertensive combination therapy as a model

The present study was designed to develop and validate a simple and fast RP-HPLC method for si-multaneously evaluating hydrochlorothiazide (HCTZ), amlodipine besylate (AML), and losartan potassium (LOS). An Eclipse plus C18 column (250 x 4.6 mm, i.d. 5 mu m) is used as a stationary phase and a simple gradient elution system. The mobile phase "A" is ethanol, and mobile phase "B" is KH2PO4 (0.01 M; pH 3.5 adjusted with 1% orthophosphoric acid). From 0 to 3.2 min into the gradient elution, the ratio of mobile phase A: B is 10:90%, v/v, and from 3.2 to 12 min, the gradient elution ratio is changed into 50:50%, v/v. A photodiode array detector (PDA) demon-strates the separated peaks. The complete separation was achieved at the retention times of 4.4, 8.1 and 9.1 min for HCTZ, AML and LOS, respectively, and a universal 233.0 nm as a detection wavelength. The method's linearity, recovery, detection and quantification limits, precision, and accuracy have been statistically validated just before the successful real applications. The present study successfully investigated the degradation behavior of these medications when exposed to various stress conditions. Furthermore, the degradation products were effectively separated and identified. For the developed and reported comparable techniques, the current work presents a sustainability profile, which was proven with innovative assessment evaluation tools. The up-to-date ChlorTox Scale indicator, CALIFICAMET tool, RGB12 algorithm, Greenness Index tool through spider plots, white and green analytical chemistry principles, and green engineering con- cepts have merged. The comprehensive findings obtained from utilizing various tools and indica-tors consistently indicate that the developed method surpasses the previously mentioned ones in terms of its environmentally friendly and sustainable characteristics.