A simple, sensitive HPTLC method has been developed and validated for the quantitative estimation of andrographolide in callus extracts of Andrographis paniculata. The stationary phase used was precoated silica gel 60F 254 plate. The mobile phase used was a mixture of chloroform: toluene: methanol (6:2.5:1.5 v/v/v). The detection of spots was carried out densitometrically using a UV detector at 310 nm in absorbance mode. Andrographolide showed mean Rf value of 0.59 with λmax at 231 nm. The method was validated in terms of linearity (100 – 700 ng), precision and accuracy (99.52% recovery). Limit of detection and limit of quantification were found to be 30 ng and 100 ng respectively. The calibration curve was found to be linear between 100 to 700 ng/spot for andrographolide. The validated method was applied for quantification of andrographolide in methanolic extracts of Andrographis paniculata samples obtained from various callus tissues. The proposed method is simple, sensitive and precise; it can be used for routine analysis.
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