Isolation and Identification of Bioactive Chlorogenic Acid from Ethanol Fraction of Cordia subcordata: HPLC, HPTLC, FTIR, and LC-MS Analysis.

Abstract

Cordia subcordata was subjected for soxhlet extraction using soxhlet apparatus by hydroalcoholic solvent (Ethanol:water; 70:30). The obtained crude extract was subjected to organoleptic properties. The extract was dark greenish or greenish-black in color. The odor of the extract was strong like tamarind. The extract taste was bitter mixed with pungent and the texture was sticky solid. The yield of the extract was 9.1%. It was observed that crude extract contains carbohydrates, reducing sugars, monosaccharides, proteins, amino acids, fats and oil, steroids, cardiac glycosides, saponin glycosides, alkaloids, tannins and phenolic compounds, and flavonoids. The presence of many phytochemicals in the extract signifies its substantial contribution to various disorders. The crude extract was subjected to column fractionation using various solvents. For column fractionation, we used polar solvents and analyzed the resulting fractions using thin-layer chromatography (TLC) to determine the presence of individual or multiple compounds. From TLC analysis, it was observed that the ethanol fraction contains a single compound and therefore same fraction was subjected for HPLC, HPTLC, FTIR, and LC-MS analysis. From HPLC analysis, it was clearly identified that this ethanolic fraction contains chlorogenic acid and it displayed a peak at 8.428 minutes. The HPTLC analysis revealed the presence of chlorogenic acid. From HPTLC analysis, chlorogenic acid was identified at 0.65 Rf value. From the present investigation, we concluded that this developed method can be used for the isolation of chlorogenic acid from C. subcordata.