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Jul 9, 2021Keywords:
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Main Article Content
Evaluation of in vitro cytotoxicity effect of Clinacanthus nutans (Brum. f.) Lindau standardized leaf extracts
Abstract
Purpose: To standardize Clinacanthus nutans (CN) leaf extracts, evaluate their contents of orientin, vitexin and isovitexin using a reversed-phase high-performance liquid chromatography (RP-HPLC) method, and also to investigate in vitro cytotoxicity of CN.
Methods: CN leaf powder was macerated in distilled water, methanol, methanol (50 %), ethanol, and ethanol (50 %) over a hot water bath at 50 - 55 °C for 24 h. The extracts were standardized for total phenolic, flavonoid, proteins and polysaccharides content by ultra-violet (UV) spectrophotometry. Moreover, RP-HPLC was used to determine the contents of orientin, vitexin and isovitexin in the extracts. The anti-proliferative effect of the extracts against human colorectal carcinoma cell line (HCT116) and human colon normal cell line (CCD-18Co) was assessed using 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl tetrazolium bromide (MTT) assay. The most active extract was fractionated using silica gel flash column chromatography to produce 20 fractions. All the fractions were subjected to the MTT test.
Results: The extracts showed modest cytotoxicity against HCT-116 and non-cytotoxicity against CCD18Co cell lines. Of all the extracts tested, the methanol extract (CN-M) showed the highest activity of all the extracts and had the highest content of flavonoid and phenolic compounds. Twenty fractions were obtained from this extract. Fraction nos. F3, F4, F14 and F16 showed significant (p < 0.05) cytotoxicity against HCT-116, with F14 having the highest activity.
Conclusion: Fraction F14 has the potential to be developed to anti-colon cancer agent. However, further studies including chemical profiling, mechanism of action and safety profile of this fraction are required.
