Detection of Residues of Cardenolides of Nerium oleander by High-Performance Thin-Layer Chromatography in Autopsy Samples

Background: Nerium oleander is an evergreen shrub of Apocynaceae family cultivated worldwide as an ornamental plant. All parts of the plant are toxic and contain a mixture of very toxic cardiac glycosides of cardenolides. A number of techniques were used to determine the cardenolides of N.oleander in various biological matrices. A survey of literature has revealed that the use of high-performance thinlayer chromatography (HPTLC) for the detection of oleander glycosides is very scanty. Method: A simple and rapid HPTLC method for separation and identification of cardenolides of N.oleander is reported. The cardenolides present in the aerial parts of the plant and residues available in the autopsy samples sent in cases of poisoning; were extracted with chloroform by using accelerated solvent extractor (ASE). Results: Separation of cardenolides was achieved on precoated silica gel 60F254 HPTLC plates with chloroform-acetone-acetic acid 8.5:1:0.5 (v/v) as mobile phase and densitometric analysis was carried out at 275 nm. A comprehensive study for the separation and detection of cardenolides in general and oleandrin in particular were studied by new mobile phases and spray reagents. The 1H-NMR spectra were recorded for the separated components and the component corresponding to oleandrin was identified. Conclusion: The method has specific advantages that it is simple, rapid and has higher resolution of separation achieved so as to be free from interferences from the plant and forensic matrices.