Abstract
A new GLC-MS method for the separation and identification of cardenolide acetates in hydrolyzed plant extracts was established. The method was shown to be selective and linear in the range from 10 μg to 5 mg/ml. Detection limits in a selected ion modus (SIM) were determined to be 5 μg/ml for digitoxigenin acetate, digoxigenin diacetate and diginatigenin triacetate and 1 μg/ml for gitoxin diacetate. Recovery rates were determined to be 96%. MS-fragmentation revealed that cardenolides with different hydroxylation patterns can be differentiated by using distinct mass fragments as indicators. While 12 β-hydroxylated cardenolides are characterized by m/z 201 and 229, cardenolides without this function fragment mainly to m/z 203 and 231. Δ5,6-unsaturated cardenolides (xysmalogenin acetate) or 5α-cardenolides (uzarigenin acetate) show fragmentation patterns similar to those of saturated or 5β-cardenolides.
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