ISOLATION AND QUANTIFICATION OF TANNINS FROM THE ROOT BARK OF CLERODENDRUM INFORTUNATUM LINN. AND ASSESSMENT OF THEIR ANTIOXIDANT POTENTIAL AND ANTIPROLIFERATIVE EFFECT ON HCT-15 CELLS
Keywords:
Clerodendrum infortunatum, Verbenaceae family, Hydrolysable tannins, Antiproliferative effect, Colon cancer, Thin layer chromatographyAbstract
Objectives: Clerodendrum infortunatum is a traditional Indian medicinal plant which has been used for the treatment of many diseases. The root bark juice is effective against indigestion and abdominal pain. Based on the preliminary screening study on the antioxidant activity of the plant, the present study was aimed to isolate tannins from the root bark of Clerodendrum infortunatum and it was qualitatively analysed and quantified. The in-vitro antioxidant activity and antiproliferative effect on HCT-15 cell lines was also evaluated.
Methods: Tannins were isolated from the root bark of Clerodendrum infortunatum. It was qualitatively analysed by phytochemical screening, protein precipitation test and thin layer chromatography (TLC) and quantified by Folin-denis assay and protein precipitation assay. The modified method of protein precipitation assay was carried out to differentiate between condensed and hydrolysable tannins. Antioxidant and antiproliferative activity were also evaluated.
Results: Phytochemical screening revealed the presence of tannins. TLC and protein precipitation ability of the extract confirmed the presence of tannins. The total tannin content by Folin-denis assay was found to be 166.6±5.607 mg tannic acid equivalents/g dry extract. This coincides with the radial diffusion method (163.75±7.5 mg tannic acid equivalents/g dry extract). Isolated tannins exhibited significant antioxidant and radical scavenging activities compared to the standard antioxidant ascorbic acid. 3-(4, 5 dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay demonstrated the antiproliferative effect of isolated tannins against HCT-15 cell lines and about 50% of the cells were found to be dead at a concentration of 100 µg/ml.
Conclusion: The antioxidant and antiproliferative properties exhibited by tannins isolated from Clerodendrum infortunatum suggest it as a new source of medicament.
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