oleandrin and oleandrigenin

An unusual case of poisoning by the ingestion of oleander leaves is reported. A 71 year old male laboratory technician committed suicide at home in this unusual manner. At the death scene a steel pan and other paraphernalia, used for the extraction of oleandrin and other cardiac glycosides from the leaves of the Nerium oleander plant were found.Toxicological investigations for oleandrin, oleandrigenin, neritaloside, and odoroside were performed by LC-MS/MS on all biological samples (peripheral blood, vitreous humor, urine, liver, gastric contents) and on the yellow infusion found at the death scene.In all samples, toxic levels of oleandrin were detected (blood 37.5 ng/mL, vitreous humor 12.6 ng/mL, urine 83.8 ng/mL, liver 205 ng/mg, gastric content 31.2 µg/mL, infusion 38.5 µg/mL). Qualitative results for oleandrigenin, neritaloside, and odoroside were obtained. Oleandrigenin was present in all tissue samples whereas neritaloside and odoroside were absent in the blood and vitreous humor but present in urine, liver, gastric content, and in the leaf brew.The purpose of this study was the identification of oleandrin and its congener oleandrigenin, detected in the vitreous humor. The blood/vitreous humor ratio was also calculated in order to assess of the likely time interval from ingestion to death. According to the toxicological results death was attributed to fatal arrhythmia due to oleander intoxication. The manner of death was classified as suicide through the ingestion of the infusion.

Topics: Aged; Cardenolides; Gastrointestinal Contents; Humans; Liver; Male; Nerium; Plant Leaves; Suicide, Completed; Vitreous Body

An HPLC/MS/MS method has been developed for the characterization and quantification of the cardiac glycosides oleandrin, odoroside, neritaloside and the aglycone oleandrigenin, all contained in a patented-hot-water extract of Nerium oleander L (Anvirzel). Qualitative analysis of such extracts was achieved using a hybrid tandem quadrupole time-of-flight (QqTOF) mass spectrometer. Collision-induced dissociation (CID) mass spectra of oleandrin, oleandrigenin, odoroside, and neritaloside were obtained with greater than 5 ppm mass accuracy and resolution routinely in excess of 8000 (fwhm). The detection limit for oleandrin of 20 pg (injected) was realized when the precursor-to-product ion transition, m/z 577 --> 373, was monitored. We have also applied the analytical method to the determination of oleandrin, oleandrigenin, neritaloside, and odoroside in human plasma following an intramuscular injection of Anvirzel.

Topics: Antineoplastic Agents; Cardenolides; Cardiac Glycosides; Chromatography, High Pressure Liquid; Humans; Mass Spectrometry; Molecular Structure; Plant Extracts; Plants, Medicinal

The purpose of this study was to examine the mechanism(s) and differential cell-killing effects of Anvirzel, an extract of oleander (Nerium oleander; family-Apocynaceae), and its derivative compound Oleandrin on human, canine and murine tumor cells. Cells received different concentrations of Anvirzel (1.0 ng/ml to 500 microg/ml) or Oleandrin (0.01 ng/ml to 50 microg/ml) in both continuously treated and pulse-treated/recovery cultures. The cytotoxicity of these compounds was then determined. Both Anvirzel and Oleandrin were able to induce cell killing in human cancer cells, but not in murine cancer cells; the cell-killing potency of Oleandrin was greater than that of Anvirzel. Canine oral cancer cells treated with Anvirzel showed intermediate levels of response, with some abnormal metaphases and cell death resulting from the treatment. From these results we conclude that Anvirzel and Oleandrin act in a species-specific manner, and while testing the effectiveness of a new compound for cancer treatment, one must use not only murine but a variety of cancer cells, including those of human origin.

Topics: Animals; Antineoplastic Agents; Blotting, Western; Cardenolides; Cell Death; DNA-Binding Proteins; Dogs; Dose-Response Relationship, Drug; Flow Cytometry; Humans; In Situ Hybridization, Fluorescence; Male; Melanoma, Experimental; Metaphase; Mice; Plant Extracts; Plants; Prostatic Neoplasms; Species Specificity; Telomeric Repeat Binding Protein 2; Tumor Cells, Cultured

Oleandrin plant poisoning is common in children and the plant extract is used in Chinese medicines. The toxicity is due to oleandrin and the deglycosylated metabolite oleandrigenin. Bufalin and cinobufotalin (toad cardiac toxins) are also widely used in Chinese medicines like Chan SU, and Lu-Shen -WU. Severe toxicity from bufalin after consumption of toad soup has been reported. Taking advantage of structural similarities of these toxins with digitoxin, we demonstrated that these compounds can be rapidly detected in blood by the fluorescence polarization immunoassay for digitoxin. The cross reactivities of these compounds with digoxin assay were much lower. For example, when a drug free serum was supplemented with 10 microg/ml of oleandrin, we observed 127.7 ng/ml of digitoxin equivalent but only 2.4 ng/ml of digoxin equivalent concentration. Digibind neutralized all cardiac toxins studied as evidenced by significant fall of free concentrations. When aliquots of serum pool containing 50.0 microg/ml of oleandrin were supplemented with 0, 10.0, 25.0, 50.0, 100, and 200 microg/ml of digibind, the mean free concentrations were 30.6, 23.3, 16.0, 10.7, 7.8 and 5.5 microg/ml respectively. Similarly, with 50.0 microg/ml of oleandrigenin (total concentration: 36.2 ng/ml), the free concentration was 14.5 ng/ml digitoxin equivalent in the absence of digibind and 5.4 ng/ml in the presence of 200 microg/ml of digibind. In another specimen containing 500 ng/ml bufalin (total concentration: 156.9 ng/ml), the free concentration was 8.6 ng/ml in the absence of digibind and none detected in the presence of 100.0 microg/ml digibind. Because such neutralization may also occur in vivo, digibind may be useful in treating patients exposed to these toxins.

Topics: Bufanolides; Cardenolides; Cardiotonic Agents; Chromatography, High Pressure Liquid; Cross Reactions; Digitoxin; Digoxin; Humans; Immunoassay; Immunoglobulin Fab Fragments; Mass Spectrometry; Neutralization Tests

Toxicity from ingestion of the oleander plant is common. Oleandrin, the oleander glycoside, has structural similarity to cardiac glycoside digoxin and is known to cross react with various digoxin immunoassays. The authors studied the cross reactivity of oleandrin and its deglycosylated congener oleandrigenin with a fluorescence polarization immunoassay for digitoxin and compared their results with a new chemiluminescent assay for digitoxin on the Automated Chemiluminescent System (ACS:180 Plus) from Chiron Diagnostics. Even though the chemiluminescent assay has been reported to be comparable with the fluorescence polarization assay among normal patient population, oleandrin and oleandrigenin showed very high cross reactivities with the fluorescence polarization immunoassay and minimal cross reactivity with the new chemiluminescent assay. When the authors supplemented a serum specimen containing no digitoxin with 50 micrograms/ml of oleandrin, the fluorescence polarization assay recorded a value of 535.7 ng/ml of digitoxin equivalent, whereas the new chemiluminescent assay recorded a value of 10.3 ng/ml of digitoxin equivalent. The cross reactivity of oleandrigenin with the fluorescence polarization immunoassay for digitoxin was significantly lower than oleandrin. The presence of oleandrin also falsely elevated total digitoxin level in a specimen supplemented with digitoxin and oleandrin. The authors also measured free digitoxin concentration by the fluorescence polarization immunoassay in the ultrafiltrate of serum supplemented with digitoxin and oleandrin. Because digitoxin and oleandrin are bound strongly to protein, monitoring free digitoxin concentration by the fluorescence polarization immunoassay instead of total digitoxin concentration does not eliminate oleandrin interference. The authors conclude that fluorescence polarization immunoassay for digitoxin has a high cross reactivity with oleandrin and can falsely elevate digitoxin concentration in the presence of oleandrin, whereas the new chemiluminescent assay for digitoxin is almost free from interferences from oleandrin.

Topics: Anti-Arrhythmia Agents; Cardenolides; Cross Reactions; Digitoxin; Fluorescence Polarization Immunoassay; Immunoassay; Luminescent Measurements

A case is presented of cardiac glycoside poisoning in a 1-year-old patient from the plant Nerium oleander (common oleander). The patient had bradycardia, vomiting, altered level of consciousness, and no history of ingestion. Antibody-based digoxin assays may cross-react with other cardiac glycosides nonquantitatively. Chromatographic techniques can be used in the specific diagnosis.

Topics: Animals; Anti-Arrhythmia Agents; Bradycardia; Cardenolides; Chromatography, High Pressure Liquid; Cross Reactions; Digoxin; False Positive Reactions; Glycosides; Humans; Immunoassay; Infant; Male; Plant Poisoning; Vomiting

Ingestion of oleander plant, containing the cardiac glycoside oleandrin, has been reported to induce fatal poisonings. Derivatives of oleandrin are structurally similar to digoxin. We investigated the cross-reactivities of oleandrin and its aglycone metabolite, oleandrigenin, in several commercially available digoxin immunoassays; assessed their ability to inhibit Na,K-ATPase catalytic activity; and measured their binding to proteins in serum. As assayed with ACS:180, Stratus, RIA, On-Line, and TDx digoxin assays, oleandrin at 100 micromol/L in digoxin-free serum gave apparent digoxin values of 0, 0.83, 2.24, 2.37, and 5.34 nmol/L, respectively, whereas oleandrigenin at that concentration gave results of 0, 0.52, 0.77, 4.94, and 1.40 nmol/L. Study of Na,K-ATPase inhibition showed IC50 values (micromol/L) of 0.22 for ouabain, 0.62 for oleandrin, 1.23 for oleandrigenin, and 2.69 for digoxin. At 25 degrees C, 96% of oleandrin and 48% of oleandrigenin were bound to serum proteins. Because detection of oleandrin and oleandrigenin by digoxin immunoassays is variable between assays as well as between congeners, assessment of cross-reactivity is warranted for each assay. The inhibition of Na,K-ATPase by oleandrin and oleandrigenin confirms that they likely exert their toxic effects through inhibition of sodium pump activity. In cases of digitalis-like poisoning with suspicion of oleander ingestion, a combination of digoxin immunoassays may be useful to effectively rule out the presence of oleander.

Topics: Blood Proteins; Cardenolides; Cardiac Glycosides; Digoxin; Enzyme Inhibitors; Humans; Immunoassay; Sensitivity and Specificity; Sodium-Potassium-Exchanging ATPase