Abstract

The toxic components of the seed oil of Croton tiglium L. (Euphorbiaceae), croton oil, are known to be cocarcinogenic phorbol-12, 13-diesters and cryptic cocarcinogenic phorbol-12, 13, 20-triesters. For a rapid isolation of mixtures of these phorbol esters from the oil rotation locular countercurrent chromatography (RLCC) was used. The eluted fractions were analysed by thin layer chromatography (TLC) and 1H-nuclear magnetic resonance spectroscopy (1H-NMR). The phorbol diesters were separated from the triesters although no baseline separation could be achieved, and the diesters as well as the triesters were completeley extracted from the oil. In a simple methanolic extraction procedure, on the contrary, the phorbol triesters were partially lost. A qualitative as well as a quantitative physico-chemical evaluation method was developed for the phytochemical screening of these cocarcinogenic and cryptic cocarcinogenic compounds in biological material using 1H-NMR spectroscopy. It was possible to quantify the phorbol esters in mixtures, in semi-purified samples, or even in the complete oil, and to determine at the same time the proportion of diand triesters. Because of its speed and accuracy, this technique provides a valuable alternative for the less versatile biological methods, to screen for known classes of compounds (like e. g. phorbol esters).

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