HPTLC Based Screening of -Sitosterol from Andrographis paniculata

Sterols are secondary metabolites of plants known for their inhibitory effects on cancer cell growth, lower cholesterol and enhance immunity. b-sitosterol belongs to phytosterol which enhances antioxidant enzymes and thus reduces the oxidative stress. In present experiment, b-sitosterol was detected from Andrographis paniculata belonging to family Acanthaceae by using HPTLC. The plant is popularly known as Kalmegha having multiple pharmacological properties used for treatment of several diseases. Leaf, stem and root extracts prepared in chloroform, methanol and petroleum ether were used for detection of b-sitosterol from the plant. The chromatography was performed on TLC plates coated with Al silica gel 60 F254 and solvents used for mobile phase was toluene:ethyl acetate:formic acid (15:4.5:1.5). After development, the plates were derivatized with 10% methanolic sulphuric acid, scanned and quantified at 510 nm. The results showed the presence of b-sitosterol in all the parts with Rf value 0.62. Calibration curve was prepared and the amount of b-sitosterol was quantified in the extracts by comparing the respective peak areas with that of the standard. The correlation coefficient for b-sitosterol against reference sample was found significant (r= 99.769057%) for the concentration range of 0.5 to 4.0 µg. Leaf methanol extract showed the highest concentration of b-sitosterol, i.e., 147.6 µg/mL. The extraction efficiency of b-sitosterol was found higher in methanol followed by petroleum ether and chloroform extract. Thus, our finding shows that Andrographis paniculata has a significant concentration of b-sitosterol, which may be useful for pharmacological application against cancer, hypercholesterolemia, inflammation and for angiogenesis process.