Extraction, Isolation and Fingerprinting of Phenolic Compound from Hydroalcoholic Extract of Cordia macleodii Hook Bark

Natural products in plants extract are usually found in complex form with numerous phytoconstituents of different polarities and thus it becomes big challenge to isolate them into pure components. In addition to fingerprinting of isolated component by HPLC and spectroscopic methods, an effort is made to develop applicable column chromatographic approach for the separation and detection of phenolic compound from the fraction of hydroalcoholic extract of Cordia macleodii bark (CMB). Hydroalcoholic extract (60:40) of Cordia macleodii bark was prepared by cold maceration method and partitioned into different fractions based on increasing polarity of solvents using separating funnel. Most productive ethanolic fraction thus obtained was undertaken for column chromatographic study using ethyl acetate: methanol (20:80) as mobile phase. HPLC study performed for identification of bioactive compound and diverse spectral studies for structural elucidation. Yield of ethanolic fraction obtained from CMB hydro alcoholic extract was found to be 84.33% w/w. HPLC data recognized gallic acid in fraction since the RT of fraction matches with standard gallic acid RT of 5.080. By using spectroscopic techniques like UV, IR, 1H & 13C NMR, and MS, the structure of the isolated compound was identified as 3,4,5 trihydroxy benzoic acid, gallic acid. Ethyl acetate: methanol (20:80) is best suited mobile phase for separation and isolation of phenolic compound from the complex mixture of fraction of extract. Gallic acid confirmed by chromatographic and spectroscopic analysis may support the pharmacological relevance of its use and substantiate traditional benefits. Keywords: Hydroalcoholic extract, Cordia macleodii bark, HPLC, Spectroscopic methods, gallic acid.