Reversed-Phase High-Performance Liquid Chromatography Quantification, Gas Chromatography-Mass Spectrometry Study and In Vitro Antioxidant Potential of Clerodendrum serratum Linn. Roots

Abstract

The present study was aimed to identify and to quantify the phytoconstituents by gas chromatography-mass spectrometry, reversed-phase high-performance liquid chromatography techniques and further to evaluate their in vitro antioxidant potential. In this study, the roots of Clerodendrum serratum Linn. was subjected to soxhlet extraction using n-hexane, chloroform, ethyl acetate and ethanol solvents and the obtained extracts (CSRH, CSRC, CSREA and CSRA) were analyzed by gas chromatography-mass spectrometry technique. The caffeic acid and beta-sitosterol were quantified from the extracts using reversed-phase high-performance liquid chromatography technique. Total phenolic and flavonoid content were determined using established methods viz. Folin-Ciocalteu and aluminium chloride colorimetric assay. Antioxidant potential was assessed using in vitro methods such as 2,2-diphenyl-1-picrylhydrazyl scavenging activity; ferric reducing ability of plasma assay and total antioxidant activity. The gas chromatography-mass spectrometry analysis revealed the presence of 48 compounds, out of which 41 compounds were identified and among them caffeic acid and beta-sitosterol were quantified using reversed-phase high-performance liquid chromatography technique. The abundance of phenolic and flavonoid content was found in CSREA and CSRA fractions. The half maximal inhibitory concentration values of 2,2-diphenyl-1-picrylhydrazyl assay was found to be 1.24, 3.36, 10.61 μg/ml for the CSRA, CSREA and CSRC fractions respectively and the total antioxidant activity was found to be 960±0.25, 766±0.14 and 816.6±0.28 μg/mg for the CSRC, CSREA and CSRA fractions respectively showing strong antioxidant activity. The ferric reducing ability of plasma assay showed strongest reducing power for CSRA and CSREA fractions. The results showed that the CSRA and CSREA fractions had strong antioxidant potential. It may be concluded from the results that the Clerodendrum serratum Linn. roots possesses significant antioxidant potential may be due to presence of potent phenolic and flavonoid compounds and these finding supports the traditional use of plant to treat the oxidative stress and related disorders.