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Abstract
In this article, simultaneous quantitative estimation of two biologically active triterpenoid compound Lupeol and a steroid compound, Stigmasterol, in Costus igneus stems was performed using high-performance thin-layer chromatography (HPTLC). TLC aluminum plates precoated with silica-gel 60 F254(20 cm × 10 cm) were used with a mobile phase of n-Hexane: Ethyl acetate (80:20v/v) for lupeol, Toluene: Acetone: Acetic acid (8.9: 0.9: 0.2 v/v/v) for stigmasterol and densitometric determination of these compounds was carried out at 366 nm in reflectance/absorbance mode. The linear regression data for the calibration plots showed a good linear relationship with r = 0.99794 and r = 0.99291 for Lupeol and Stigmasterol, respectively. A recovery study was carried out to check the accuracy of the method. The average recovery of Lupeol and Stigmasterol was 100.16% and 99.94%, respectively. According to International Conference on Harmonization (ICH) guidelines, the method was validated for linearity, precision, accuracy, and recovery, limit of detection, limit of quantification, specificity, robustness, and ruggedness. The HPTLC method was found to be reproducible, accurate, and precise and could detect these two compounds at nanogram level from the stem of Costus igneus.
ACKNOWLEDGMENT
The authors acknowledge DBT, New Delhi (BT/PR10018/NNT/28/2007) for providing financial support and K. Manjula acknowledges DBT for Junior Research Fellowship, Dr. S. John Britto, Director, Rapinat herbarium, St. Joseph College, Tiruchirapalli, and Tamil Nadu for identifying the plants. The authors acknowledg Dr. N. Ramachandran, Vice-Chancellor of Periyar Maniammai University for his constant support of this research.