A VALIDATED ISOCRATIC RP-HPLC METHOD FOR CONCURRENT ESTIMATION OF GYMNEMAGENIN, GALLIC ACID AND 18Î’-GLYCYRRHETINIC ACID IN POLYHERBAL FORMULATION
Keywords:
Gymnemagenin, Gallic acid, Nil, Isocratic HPLC, ICHAbstract
Objective: To develop and validate a simple, precise, selective, and accurate reversed phase high performance liquid chromatography method for concurrent analysis of gymnemagenin, gallic acid and 18β-glycyrrhetinic acid in polyherbal formulation.
Methods: The chromatographic separation was achieved on a Thermo Synchronis C18, 5 μm, 250 × 4.6 mm i. d. analytical column. The mobile phase comprised of methanol: water (88: 12, v/v), pH 3.1 adjusted with orthophosphoric acid. The flow rate was kept at 0.8 mL min-1. Quantitation was achieved with UV detection at 218 nm, based on peak area.
Results: The retention time for gallic acid, gymnemagenin, and 18β-glycyrrhetinic acid was found to be 3.08, 4.15, and 10.30 min, respectively. Validation of the RP-HPLC method was performed as per International Conference on Harmonization (ICH) Q2 (R1) guideline. The proposed method showed good linearity in the range of 100-1000 μg mL-1 for gymnemagenin, 2.5-50 μg mL-1 for gallic acid and 50-500 μg mL-1 for 18β-glycyrrhetinic acid. The % content of gymnemagenin, gallic acid and 18β-glycyrrhetinic acid in the marketed formulation was found to be 0.1320, 0.2129 and 0.2799 %, respectively.
Conclusion: The proposed method can be useful in the quality control of gymnemagenin, gallic acid and 18β-glycyrrhetinic acid in polyherbal formulation.
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References
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