RP-HPLC and Spectrophotometric Determination of Rutin Trihydrate, Berberine Chloride and Trigonelline Hydrochloride in Antidiabetic Polyherbal Formulations
Introduction: High performance liquid chromatographic method and Spectrophotometric method by Simultaneous equation were developed for determination of three herbal markers- Rutin Trihydrate (RUT), Berberine Chloride (BER) and Trigonelline Hydrochloride (TRG) in their combined dosage form. Methods: RP-HPLC separation was achieved on Inert sustainswift, C18 column (250mm x 4.6mm i.d., 5μ) using mobile phase composed of acetonitrile: methanol: water [(15:15:70 v/v/v), pH adjusted to 3.5 with 0.1% ortho phosphoric acid) with a flow rate of 1.0 ml/min. RP-HPLC quantitation was achieved with UV detection at wavelength 270 nm. For spectrophotometric method, 249 nm, 261 nm and 274 nm were selected for determination of RUT, BER and TRG respectively. Results: The calibration curves follow Beer’s law in the range of 50-250 μg/ml in RP-HPLC and 10-30 μg/ml in spectrophotometric method for RUT, BER and TRG. Correlation coefficients were found to be close to 0.999 for three drugs. The accuracy was in the range of 99.06-102.27% for all three markers. Conclusion: The proposed methods were successfully applied for the determination of RUT, BER and TRG in pure form and in Antidiabetic Polyherbal formulations.