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

Yıl 2020, Cilt: 3 Sayı: 1, 50 - 63, 29.03.2020

Raju Asirvatham Aparna Ann Mathew Salwa Abdul Salam

Öz

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. 

Anahtar Kelimeler

Indian Drosera species, HPTLC, Plumbagin

Kaynakça

• Bag GC, Grihanjali Devi P, Bhaigyabati TH (2015). Assessment of total flavonoid content and antioxidant activity of methanolic rhizome extract of three Hedychium species of manipur valley. Int J Pharm Sci Rev Res 30(1): 154-159.
• Balkwill, Mantovani A (2001). Inflammation and cancer: back to Virchow? The Lancet 357(9255): 539–545.
• Chien-Fu H, Shih-Shen L, Pao-Hsin L, Young SC, Yang CC (2008). The immunopharmaceutical effects and mechanisms of herb medicine. Cellular and Molecular Immunology 5(1):23–31.
• Gandhidasan R, Thamaraichelvan A (1991). Anti-inflammatory action of Lannea coromandelica by HRBC membrane stabilization. Fitoterapia 62(1): 81–83.
• Jayaram K, Prasad MNV (2006). Droser aindica L. and D. burmanii Vahl., medicinally important insectivorous plants in Andhra Pradesh –regional threats and conservation. CurrSci 91:943-946.
• Madhavan V, Basnett H, Kumar AC, Yoganarasimhan SN (2008). Fingerprinting of plumbagin in Drosera burmannii Vahl using high performance thin layer chromatography. Indian J Pharm Sci 70:798-800.
• Prasanna BG, Sameera RS, Ira TG, Weerasinghe AK, Mayuri GT, Ratnasooriya WD, Kamani HT (2012). Anti-inflammatory activity is a possible mechanism by which the polyherbal formulation comprised of nigella sativa (seeds), Hemidesmusindicus (Root), and Smilax glabra (Rhizome) mediates its antihepatocarcinogenic effects. Evidence Based Complementary and Alternative Medicine: 1-12.
• Raju A , Christina AJM , Mayakrishnan A (2012). Antitumor potential of ethanol and aqueous extracts of Drosera burmannii Vahl against Dalton’s ascitic lymphoma bearing mice. J Pharm Res 5(3): 1418-1423.
• Santapau H, Henry AN (1976). A Dictionary of the flowering plants in India. Publication and information Direcorate, pp.58, New Delhi, India.
• Suresh Kumar Karri, Angappan Sheela (2017). Comparative in vitro Antidiabetic and Immunomodulatory evaluation of standardized five select medicinal herbs and spectral analysis of i L. (Nyctaginaceae). PharmacognJ 9(3): 336-344.
• Wagner H, Bladt S, Zgainski EM (1984). A Thin Layer Chromatography Atlas. In: Plant Drug Analysis – 2nd edn, pp. 276,Springer: Berlin, Germany.
• Yamada Y, Webber EM, Kirillova I, Peschon JJ, Fausto N (1998). Analysis of liver regeneration in mice lacking type 1 or type 2 tumor necrosis factor receptor: requirement for type 1 but not type 2 receptor. Hepatology 28(4I): 959–970.
• Yu He, Zhimin He, Feng He, Haitong Wan (2012). Determination of quercetin, plumbagin and total flavonoids in Droserapeltata Smith var. glabrataY.Z.Ruan. Pharmacogn Mag 8(32): 263–267.