Analytical studies on the active constituents in crude drugs. III. High-speed liquid chromatographic determination of ecdysterone and inokosterone in Achyranthis radix.

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Abstract
A simple, rapid, and accurate high-speed liquid chromatographic method was established for the determination of ecdysterone (I) and inokosterone (II) in Achyranthis radix. I and II, extracted from Achyranthis radix, were directly injected onto column and separated in approximately 15 min using a 2 m Permaphase ETH column with a mobile phase of 10% ethanol in n-hexane at 50°. A working relative standard deviation was less than 2%. The extraction methods were also investigated and it was concluded that the Soxhlet extraction with methanol was the most effective. Moreover, II was separated into two epimeric isomers on a Permaphase octadecyl silane column with a mobile phase of methanol in water.