Determination of oleander glycosides in biological matrices by high-performance liquid chromatography

A high-performance liquid chromatography (HPLC)-fluorescence method was developed for quantitative determination of oleandrin in gastrointestinal contents (stomach, rumen, colon, and cecum contents). Oleandrin was extracted with methylene chloride. The extract was cleaned up on a charcoal solid phase extraction (SPE) column, derivatized with 1-naphthoyl chloride, and quantified by HPLC with a fluorescence detector. The limit of detection was 0.05 ppm. Six replicate fortifications of stomach contents matrix at 0.10 ppm oleandrin gave an average recovery of 85%, with 4.6% CV (relative standard deviation). The diagnostic utility of the method was tested by analyzing samples submitted to the veterinary toxicology laboratory.