Identification and Quantification of Germacrene D from the Extracts of Zanthoxylum ovalifolium Leaves by AgNO3-TLC Densitometric Method

Germacrene D an important sesquiterpenoid has been extracted from leaves of Zanthoxylum ovalifoilum and high performance thin-layer chromatographic method has been developed for its simultaneous identification and determination. The compound was extracted by cold extraction and separated and identified on a AgNO3 impregnated silica gel 60 F254 HPTLC plates with dichloromethane:acetone (9.8:0.2 v/v) as mobile phase. The quantification of geremcare D was carried out by densitometry scanning performed at λ = 254 nm by reflectance scanning. This facilitated a well-resolved band for the compound with a Rf value of 0.46 ± 0.01. Linear regression concentration analysis data for mean calibration plots showed good linear relationship in the range of 1000-10000 ng/mL, with a correlation coefficient, slope and intercept of 0.9985 ± 0.0004, 21.62 ± 0.435 and 3308.87 ± 10.24, respectively. The minimum amount of germacrene D that could be authentically detected (LOD) and quantified (LOQ) were determined and found to be 212.07 and 521.23 ng/spot, respectively. The method was validated by considering the parameters linearity, precision, accuracy, specificity and robustness wherein, it is evident from the results that the proposed method was simple, safe and reliable.