PJB-2019-835
IDENTIFICATION AND CHARACTERIZATION OF ACTIVE INGREDIENT EUGENOL FROM SYZYGIUM AROMATICUM (CLOVE OIL) THROUGH HPLC AND ITS PHYTOCHEMICAL ANALYSIS
DR. SABA IRSHAD
Abstract
Background: Clove is one of the most ancient and premium sort of spice having a wide range of biological activities that makes it highly valuable in pharmaceutical and food sector. Methods: Clove oil was extracted by using Soxhlet method. Phytochemical screening was carried out by different chemical tests. DPPH scavenging assay was used to analyze antioxidant property of clove oil. For the Qualitative estimation of active components, Bromine, KMnO4 and iron chloride tests were performed respectively. Antimicrobial activity was confirmed by Disc diffusion and Well diffusion assays. Quantification of eugenol in clove oil was done by HPLC. Results: Phytochemical analysis of aqueous extract of clove showed the presence of carbohydrates, flavonoids, coumarins, steroids, saponnins, and tannins in clove while proteins and leucoanthocyanins were absent in it. In DPPH scavenging assay, an increase in percentage inhibition was observed with an increase in concentration of clove oil. Antimicrobial activity of clove oil was tested against three gram positive strains i.e. Bacillus subtilis, Staphylococcus aureus and Bacillus thuringiensis and one gram negative strain i.e. Escherichia coli by well diffusion and disc diffusion methods. T-test was applied for statistical analysis of antimicrobial activity. A significant difference was found between well diffusion and disc diffusion method as the p-value obtained was 0.0215 which is ˂0.05. Aqueous clove nanoparticles were made and were found to have antimicrobial activity against E.coli (gram -ve) and B.subtilis (gram +ve). HPLC of Clove oil along with standard i.e. eugenol was performed. Correlation coefficient (R2) was found to be 0.973531. Quantitative estimation of eugenol showed the presence of 740 ppm eugenol in said concentration of clove sample. Conclusion: Antimicrobial, antioxidant property of clove proved it as a valuable spice in pharmaceutical and food sector.
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