RP-LC Method in Gradient mode for Combined Quantification of Metformin, Linagliptin, and Empagliflozin in Combination Tablets

It has been developed and validated an accurate, sensitive, precise, quick, and gradient reverse phase HPLC (RP-HPLC) technique for combined quantification of Metformin, Linagliptin, and Empagliflozin in raw materials and pharmaceutical combined tablets. With acetonitrile as the organic solvent, the best separation was achieved on a ۲۵۰ mm x ۴.۶ mm i.d., ۵µ-particle size Inertsil®-Octadecyl-silyl-۳V-Reverse-Phase-C۱۸-column with acetonitrile as the non-polar-modifier and Dipotassium mono-hydrogen phosphate buffers system [۰.۰۳M] in water with pH ۳.۰ adjusted with ortho-phosphoric acid (۰.۱ %v/v) in the gradient mode of elution as an elution solvent at a speed of ۱.۰ mL.min−۱. UV detection was at ۲۳۰-nm. Metformin had a retention time of ۲.۴۲۱ minutes, Linagliptin's was ۸.۱۸۷ minutes, and Empagliflozin's was ۱۱.۷۱ minutes. With a correlation coefficient of about ۰.۹۹۹۹, the peak-response was obtained as a function of concentration over the range of ۸۰-۴۸۰ mcg/mL for metformin, ۰.۴-۲.۴ mcg/mL for linagliptin, and ۰.۸-۴.۸ mcg/mL for empagliflozin. Metformin, Linagliptin, and Empagliflozin were shown to have a percentage assay of ۹۹.۹۲, ۹۹.۷۲, and ۹۹.۷۴, respectively. Metformin, Linagliptin, and Empagliflozin, each have a limit of detection of ۰.۰۴ g/mL, ۰.۰۰۴ g/mL, and ۰.۰۰۴ g/mL, respectively. Metformin, Linagliptin, and Empagliflozin, each have a limit of quantification (LOQ) of ۰.۱۲ ng/mL, ۰.۰۱۲ g/mL, and ۰.۰۱۲ g/mL, respectively. The presence of excipients in the formulation had no effect on the assay method. The procedure is appropriate for usage in QC- laboratories since it is quick and precise.

کلیدواژه ها:

Metformin ، Linagliptin ، Empagliflozin ، Gradient-RP-HPLC ، and Fixed dosage forms