Multiresidue screen for cardiotoxins by two-dimensional thin-layer chromatography.
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 III in gastrointestinal contents (stomach, rumen, colon, and cecum contents), feces, and plant material. The cardiotoxins were extracted with dichloromethane. The extract was cleaned up by charcoal and 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 by chloramine T spray. The method detection limits were 0.05 microg/g for oleandrin, 0.1 microg/g for gitoxin, and 0.2 microg/g for the other toxicants in gastrointestinal contents and feces and were 5 times higher in plant material. Four replicate fortifications of bovine rumen contents, bovine feces, and alfalfa at these levels were all well recovered. The diagnostic utility of the method was tested by analyzing samples submitted to the veterinary toxicology laboratory.
Holstege
D M
DM
Toxicology Laboratory, California Veterinary Diagnostic Laboratory System, University of California, Davis, California 95616, USA. dholsteg@cvdls.ucdavis.edu
Francis
T
T
Puschner
B
B
Booth
M C
MC
Galey
F D
FD
D000818:Animals; D002417:Cattle; D002855:Chromatography, Thin Layer / Q000379:methods*; D004179:Cobra Cardiotoxin Proteins / Q000032:analysis*; D005243:Feces / Q000737:chemistry; D000455:Medicago sativa / Q000737:chemistry; D012417:Rumen / Q000737:chemistry
D004179:Cobra Cardiotoxin Proteins