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

Abstract

Hase et al.: Antioxidant Potential of Clerodendrum serratum Linn. RootsThe 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 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 ethyl acetate and residual aqueous 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 residual aqueous fraction, ethyl acetate and chloroform 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 chloroform, ethyl acetate and residual aqueous fractions respectively showing strong antioxidant activity. The ferric reducing ability of plasma assay showed strongest reducing power for residual aqueous fraction and ethyl acetate fractions. The results showed that the residual aqueous fraction and ethyl acetate 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.

Thin layer chromatography‐densitometric method for quantification of apigenin, oleanolic acid, and β‐sitosterol in clerodendrum serratum (Bharangi) root extract

“…Leaves [12] and HPLC method for caffeic acid and β -sitosterol in C. serratum Linn. Root extract [13] has been reported. Several analytical methods have also been developed for the simultaneous determination of Oleanolic acid, β-Sitosterol, or Apigenin along with other markers in other plant and their extracts like Leptadenia pyrotechnica [14], Betula utilis D. Don [15], Syzygium Cumini [16], Nyctanthes arbor-tristis [17], and so forth.…”