Study of in vitro immunomodulatory effect and quantitative evaluation of main phytoconstituents in Indian Drosera species

Year 2020, Volume: 3 Issue: 1, 50 - 63, 29.03.2020

Raju Asirvatham , Aparna Ann Mathew , Salwa Abdul Salam

Abstract

The present study compared the total flavonoid content in ethanol and aqueous extracts of Indian Drosera and their in vitro immunomodulatory activities. None of the medicinally important compounds from this species has been quantified and compared previously. HPTLC method was used to quantify plumbagin and quercetin content in the ethanol extracts of three Indian Drosera species.
The in-vitro immunomodulatory activities of both ethanol and aqueous extracts were evaluated by the inhibition of heat and hypotonic induced hemolysis, nitroblue tetrazolium (NBT) assay and by inhibition of TNF-α release in DAL cell lines.
Ethanol extract of D. burmannii showed significant inhibition of heat and hypotonic induced hemolysis when compared with diclofenac. In NBT reduction test, ethanol extract of D. burmannii showed significantly higher reduction than D. indica and D. peltata. Inhibition of TNF-α release was significantly enhanced by 400 µg/mL of D. burmannii. Higher concentration of flavonoid was found in the ethanol extract of D. burmannii. Flavonoid concentration was the least in aqueous extract of D. indica. The calibration curve of plumbagin and quercetin were found to be linear (200 -1000 ng/spot). Correlation coefficient of r = 0.9994 ± 8.62 and r= 0.99068 ± 13.63 was detected for plumbagin and quercetin, respectively. This is indicative of good linearity between concentration and peak area.
The identity of the plumbagin and quercetin band in the sample extracts was confirmed by comparing the UV absorption spectrum of the sample to that of the reference standard plumbagin and quercetin, using the Camag TLC scanner. Higher concentrations of plumbagin and quercetin were found in the ethanol extract of D. burmannii. The proposed HPTLC method provided good resolution and accuracy, and can be practiced for the rapid determination of plumbagin and quercetin in the herbal drugs. Such an approach is effective for routine quality control analysis and quantification of plumbagin. The pharmacological actions of the species investiagted are due to its chief chemical constituents. 

Keywords

Indian Drosera species, HPTLC, Plumbagin

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