Development and validation of an isocratic HPLC method for simultaneous determination of quaternary mixtures of antihypertensive drugs in pharmaceutical formulations

Abstract

Summary. The objective of this study was to develop and validate an assay method for simultaneous determination of atenolol, furosemide, losartan, and spironolactone in pharmaceutical formulations. A reverse-phase high-performance liquid chromatography procedure was developed, using a Kinetex ® C-18 column (100 mm × 4.6 mm, 2.6 µm). The mobile phase was composed of methanol-water (75:25 v/v, pH 3.0, adjusted with phosphoric acid), with a flow rate of 0.4 mL min −1 . All drugs were separated in less than 5 min. The method was validated according to International Conference on Harmonization (ICH) and Association of Official Analytical Chemists (AOAC) guidelines. The method showed linearity in a concentration range of 0.75-12.0 µg mL −1 for atenolol (r = 0.9995), 0.30-12.00 µg mL −1 for furosemide (r = 0.9997), 0.45-12.00 µg mL −1 for losartan (r = 0.9995), and 0.45-12.0 µg mL −1 for spironolactone (r = 0.9999). The method also showed repeatability and precision. The three-day average intra-day precisions were 101.35 ± 0.74% for atenolol, 95.84 ± 1.44% for furosemide, 98.90 ± 1.16% for losartan, and 97.19 ± 0.18% for spironolactone. Similarly, the inter-day precisions were 101.34 ± 0.72% for atenolol, 95.84 ± 0.1.50% for furosemide, 98.90 ± 1.17% for losartan, and 97.19 ± 0.83% for spironolactone. The method accuracy was also tested and validated -in this case, the average recovery values were 100.18 ± 1.20% for atenolol, 99.83 ± 1.54% for furosemide, 100.07 ± 0.95% for losartan, and 99.94 ± 0.93% for spironolactone. Finally, the method was successfully applied in the simultaneous determination of atenolol, furosemide, losartan, and spironolactone in magisterial formulas, as well as in commercial pharmaceutical formulations.

Intensification of UV-C treatment to remove emerging contaminants by UV-C/H2O2 and UV-C/S2O82−: Susceptibility to photolysis and investigation of acute toxicity

“…The separation was performed on C18 columns (Poroshell EC-C18 (3 x 50 mm, internal diameter 2.7 µm) followed by Poroshell HPH-C18 (4.6 x 150 mm, internal diameter 2.7 µm) at a flow rate of 0.5 mL min -1 . Ultra-pure water (40%) and MeOH (60%; MERCK HPLC grade, 98% purity), both containing 0.1% formic acid (SIGMA-ALDRICH HPLC grade, 98% purity), was used as the mobile phase for LP and FRSM, adapting previously described procedures [42]. Both LP and FRSM were monitored at =240 nm.…”

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“…In reviewing the literature, various analytical methods were found for the determination of atenolol whether alone or in its combination with other drugs in pharmaceutical preparations including spectrophotometry 4-6 kinetics 7, 8 Titrimetric 9 GC 10 and HPLC [11][12][13][14][15] .…”

The development of HPLC-DAD method for determination of active pharmaceutical ingredient in the potassium 2-((4-amino-5-(morpholinomethyl)-4H-1,2,4-triazol-3-yl)thio) acetate substance

“…Of these salts, The development of HPLC-DAD method for determination of active pharmaceutical ingredient in the potassium 2-((4-amino-5-(morpholinomethyl)-4H-1,2,4-triazol-3-yl)thio) acetate substance 2-((4-amino-5-(morpholinomethyl)-4H-1,2,4-triazol-3-yl) thio)acetate, which along with low toxicity, exhibits antioxidant and hepatoprotective properties [11]. Therefore, the development of a method for determination the API in the substance is one of the important steps towards the creation of a new medicinal product [12][13][14].…”