A two-dimensional thin-layer chromatographic method was developed for the qualitative determination of the cardiotoxins oleandrin, gitoxin, digitoxin, gitoxigenin, and grayanotoxins I, II, and IIIin gastrointestinal contents (stomach, rumen, colon, and cecum contents), feces, and plant material.The cardiotoxins were extracted with dichloromethane. The extract was cleaned up by charcoaland reverse phase solid-phase extraction columns. Analysis was performed by two-dimensional thin-layer chromatography on silica gel plates and visualized by aluminum chloride followed bychloramine T
spray. The method detection limits were 0.05
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g/g for oleandrin, 0.1
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g/g for gitoxin,and 0.2
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g/g for the other toxicants in gastrointestinal contents and feces and were 5 times higherin plant material. Four replicate fortifications of bovine rumen contents, bovine feces, and alfalfaat these levels were all well recovered. The diagnostic utility of the method was tested by analyzingsamples submitted to the veterinary toxicology laboratory.Keywords: Oleandrin; digitoxin; gitoxin; grayanotoxin; thin-layer chromatography