A VALIDATED HPLC METHOD FOR THE DETERMINATION OF BETULIN IN THE STEM BARK OF TECTONA GRANDIS LINN.

Betulin, a known pentacyclic tri-terpenoid, possessing diverse pharmacological properties was determined using HPLC in stem bark methanol extract of Tectona grandis Linn. distributed in greater part of India. Furthermore, Tectona grandis Linn. is used traditionally in treatment of bronchitis, dysentery, headache, leukoderma, leprosy and constipation. The present study was aimed to validate HPLC method for the determination of betulin in Tectona grandis Linn. (bark). The chromatographic separation was achieved using Waters HPLC instrument, reverse phase Hypersil C18 column (250 mm) under isocratic elution of acetonitrile-water 85:15 (v/v) with a flow rate of 1.0mL min ˉ¹ and the run time was set at 15 min. The detection was done at 210 nm and the column temperature was maintained at 25⁰ C. The method was linear for betulin over the concentration range of 28.42- 53.99 ppm (R² = 0.9974) in Tectona grandis Linn. (bark). The quantification of betulin in methanol extract of Tectona grandis Linn. was validated in terms of system precision, method precision, linearity, recovery, robustness, ruggedness, LOD and LOQ according to ICH guidelines. The % RSD values were found to be in an acceptable range as per ICH guidelines