Abstract
A stability-indicating reversed-phase liquid chromatographic (RPLC) method has been established for analysis of rutin (1), kaempferol-3-O-robinobioside (2), and nicotiflorin (3) in Desmodium gangeticum. The study was performed in the presence of the degradation products generated in the study of forced degradation. Marker compounds were subjected to stress by hydrolysis (acidic and basic), oxidation, photolysis, and thermal treatment. Under the optimized conditions, well-resolved separation of pure compounds from the degradation products with significantly different Rt values was achieved on a Spherisorb ODS2 column (250 mm × 4.6 mm, 10 µm) using isocratic elution of methanol and water (0.5% acetic acid); with acceptable validation results such as linearity, sensitivity, and recovery in terms of RSD (%). The calibration curves were linear in the concentration range of 20–100 µg/mL. Method was validated as per ICH guidelines. The reproducible and robust method may be applied for assays and stability tests of D. gangeticum and phytopreparations containing D. gangeticum.
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