Quercetin Lab Report

Abstract A natural polyphenolic compound Quercetin was extracted from widely available and consumed plant Allium fistulosum which has shown promising chemo preventive and chemotherapeutic activities in cancer. In this study, quercetin is encapsulated and loaded onto PLGA (Poly lactic glycolic acid) microspheres using double emulsification methodology for improving its efficiency in cancer therapy. Quercetin is extracted and purified using chromatographic and nuclear magnetic resonance (NMR) techniques. PLGA-heparin (PLGAh) and PLGA-heparin-quercetin (PLGAh+Q) loaded microspheres were characterized for morphology using scanning electron microscopy (FESEM). The PLGAh and PLGAh-Q microspheres were tested for its cytotoxicity against MCF-7 lines.
4. Results
4.1 Column chromatography
The column chromatography was performed using different concentrations of elution solution and the column is eluted in increasing order of polarity of solvent from 100 % of extract, 25 % of ethyl acetate in hexane, 50 % of ethyl acetate in hexane, 75 % of ethyl acetate in hexane and 100 % of ethyl acetate. Then the polarity was increased using 2 % Methanol in Ethyl acetate to 50 % Methanol in ethyl acetate. 8 % of methanol and 92 % of ethyl acetate concentration was found to be the best concentration and after concentrating all fractions of these concentrations, approximately 7 to 8 mg of quercetin was obtained.

4.2 Thin layer chromatography
TLC profiles were carried out for the fractions, performed using the solvent mixture of toluene: ethyl acetate: methanol in the ratio of 5:3:2 (v/v/v) and this mobile phase enabled excellent separation of the bioactive compound. The results showed that there was good amount of correlation between the standard quercetin band and the band of bioactive sample plates when visualized under UV at 254
The PLGAh and PLGAh+Q carrier was tested by these assay for rate of apoptosis in the in Michigan cancer foundation (MCF-7) cell lines. The control was the normal cancer cell lines without any PLGAh and PLGAh-Q carrier. The PLGAh carrier was tested in assay in two different concentrations which was found above 1.5 µg and 3.0 µg. The summation of early and late apoptosis was just 8 % in concentration of 1.5 µg and in concentration of the 3.0 µg it was found to be 15 %. But PLGAh-Q in the concentration of 1.5 µg showed approximately 45 % of apoptosis (early+ late). But when concentration was increased to the 3.0 µg the apoptosis was (early + late) found to be 60 %. So it is concluded that PLGAh carrier in various concentration (1.5 µg and 3.0 µg) does not show any significant apoptosis but the PLGAh-Q carrier showed increase in apoptotic activity by increasing the concentration of drug from 1.5 µg to 30 µg. Studies have indicate that the viability of cancer cell decreased when it was given silver nanoparticles in different concentration and the confirmation of different stages of apoptosis were observed by separating the live cell and dead cells by annexin V / PI staining which also