ABSTRACT
An LC-MS screening method was developed to detect the presence of atractyloside (ATR), the toxic principle of a commonly used medicinal plant in South Africa, Callilepis laureola, in biological matrices such as body fluids and human viscera.
Subject(s)
Atractyloside/analysis , Atractyloside/poisoning , Callilepis , Plant Poisoning/diagnosis , Viscera/chemistry , Body Fluids/chemistry , Callilepis/chemistry , Callilepis/poisoning , Chromatography, Liquid , Forensic Pathology , Humans , Plant Extracts/chemistry , Plant Extracts/poisoning , Plant Poisoning/pathology , Plant Roots/chemistry , Spectrometry, Mass, Electrospray IonizationABSTRACT
A selective analytical method based on high-performance liquid chromatography, combined with atmospheric pressure ionisation mass spectrometry, was developed for the detection of atractyloside. The analysis was performed on an Xterra Phenyl column utilising a gradient elution profile and a mobile phase consisting of 10 mM aqueous ammonium acetate buffer-methanol-acetonitrile. The calibration curve of the method (1 ng/ml-160 microg/ml) was best described by a second order polynomial function (r2 = 0.998) but displayed good linearity in the range of 100 ng/ml-1 microg/ml (r2 = 0.999). The limit of detection for the atractyloside standard was determined and found to be 100 pg/ml and the limit of quantification of atractyloside in tuber matrix was found to be 250 pg/ml. The relative standard deviation of the method was on average below 5% (n = 8). The method was successfully applied to the analysis of Callilepis laureola tubers and unknown powdered samples for the presence of atractyloside.
Subject(s)
Callilepis/chemistry , Mass Spectrometry/methods , Atmospheric Pressure , Flow Injection Analysis , Sensitivity and SpecificityABSTRACT
The traditional Zulu remedy impila (Callilepis laureola) can cause acute fatal hepatocellular necrosis, especially in children. We investigated the mechanism(s) of toxicity using HuH-7 hepatocytes. Impila tubers were extracted with boiling water and the aqueous extract was used at different concentrations to study the effects on the morphology of the cells. Flow cytometry and labelling with fluorescent antibodies to tubulin were also used. At high concentrations, necrosis occurred; however, at lower concentrations, the extracts gave rise to a variety of changes including hypercondensation of chromatin, multinucleate cells, nuclear fragmentation and apoptosis. In addition, we observed destruction of cytoplasmic tubulin. These findings give further insight into the mechanism of toxicity of herbal remedies containing atractyloside.
