Quantitative analysis of four active constituents in Tibetan herb Gymnadenia conopsea by high-performance liquid chromatography
Yang, B.; Li, S.; Zhang, R.; Wang, Y.; Shi, J.
Zhongguo Zhong Yao Za Zhi 34(14): 1819-1822
2009
ISSN/ISBN: 1001-5302 PMID: 19894516 Document Number: 629791
To develop a high performance liquid chromatography method for simultaneous determination of tour active constituents, dactylorhin B, loroglossin, dactylorhin A, militarine present in a Tibetan herb Gymnadenia conopsea. The analysiswas achieved on a Pravil C18 column (4.6 mm x 150 mm, 5 microm) eluted with the mobile phases of methanol (A)-water (B) in gradient elution. The initial condition was 15% A, chsnhrf to 20% A in 10 min, to 45% A in 10 min, to 70% A in 15 min. The detection wavelength was set at 222 nm. The flow rate was 0.7 mL x min(-1). Orthogonal test was adopted to optimize extraction process of four active compounds from Gymnadenia conopsea. The assay displayed good linearity over the concentration ranges of 0.20-4.05 microg (r = 0.9999), 0.10-5.11 micro g (r = 0.9999), 0.10-5.11 microg (r = 0.9999) and 0.09-1.79 microg (r = 0.9999), respectively. The average recoveries (n=9) were 99.19%, 99.16%, 99.52% and 99.00% for dactylorhin B, loroglossin, dactylorhin A and militarine respectively. The method is simple, sensitive, reliable and reproducible which can be used for the quality study of Gymnadenia conopsea.