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This study was designed to develop and validate a liquid chromatographic method for determination of Embelin in crude extract of Embelia ribes. The chromatographic separation was performed on a HPLC coupled to a diode array detector with Thermo Hypersil-Keystone, BDS C-18 column (25 cm L x 4.6 mm ID; particle size- 5μ) using methanol and 0.1% TFA (88:12 v/v) as mobile phase, at a flow rate of 1 ml/min. Quantification was achieved with UV detection at 288nm, based on peak area. The optimized procedure involved the extraction of Embelin in crude extract of Embelia ribes by soxhlet and maceration using chloroform and methanol as a solvent. Extraction by soxhlet at 70°C for 2 hr using chloroform as solvent was the best suited method. The assay method was developed and validated for system suitability, linearity, range, accuracy, precision and robustness. The plot of integrated peak area and concentration of Embelin was found to be linear over a range of 6.25-200 μg/ml. The LOD and LOQ (μg/ml) levels were, 1.5 and 4.5, respectively. The recovery experiments led to mean recovery of 98.6%. This indicates that this method is simple, rapid, sensitive, precise and accurate and can be applicable for the extraction and quantitative determination of Embelin in crude extracts of Embelia ribes.
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Author(s): Rastogi S Bhatia AK Kushwaha A Pandey MK Sharma A Jai Prakash Singh GN
Extract of Embelia ribes, Embelin, HPLC