HPLC Separation of Cephalotaxine, Harringtonine and Homoharringtonine from Callus and Root Cultures ofCephalotaxus Harringtonia

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Abstract
A simple extraction and analytical protocol was developed to assay cephalotaxine, harringtonine, and homoharringtonine from callus and root cultures of Cephalotaxus harringtonia. The process involves extraction by methanol followed by partitioning between 0.5% ammonium hydroxide and chloroform. The chloroform fraction recovered greater than 90% of all three alkaloids. This fraction was concentrated to dryness, resuspended in methanol and analyzed by high pressure liquid chromatography using a UV detector. Peaks corresponding to all three alkaloids were identified by comparing their retention times with authentic standards and their identity confirmed by fast atom bombardment spectrometry. The root cultures contained higher levels of harringtonine and homoharringtonine (2.4 and 3.9 mg/kg dry matter, respectively) compared to the callus cultures; however, the levels of cephalotaxine were comparable (10.2 mg/kg dry matter).