Development of HPTLC method for quantification of vasicine in marketed Herbal formulations containing Adhatoda vasica

Abstract

Leaves of Adhatoda vasica, commonly known as vasaka (family: Acanthaceae) are used as alcoholic extracts and liquid extracts or syrup for treating cold cough, whooping cough, chronic bronchitis and asthma. The concentration of vasicine, the active principle varies greatly from 0.541 to 1.105% on dry basis [1]. To analyze vasicine content of two poly-herbal formulations containing vasaka, we developed a simple, fast, sensitive, precise, and reliable HPTLC method. Precoated Silica gel 60 F 254 plates were spotted with 4–12µg/ml of vasicine and developed with methanol:toluene:dioxane:ammonia (2:2:5:1). The spots were scanned at 254nm to prepare the standard calibration curve. 10ml of the formulation was extracted with methanol and the extract upon concentration was dissolved in known volume of methanol. 10µl of this solution was used for the quantitative estimation [2]. The Vasicine content in the selected formulations were determined by comparing area of the chromatogram of formulations with the standard vasicine. The regression coefficient of calibration curve was 0.998 and the curve was linear from 4–40µg. The method was found to be precise with mean±SD of 36.298±0.020. The vasicine content of two formulations varied greatly suggesting necessity of developing a well validated method for preparation of formulations. Recovery studies were satisfactory. Thus the method was found reliable and practicable.