As the connectivity of the world continues to grow, access to both native and non-native plant materials has increased in many areas. Some of these materials are used in traditional medicines and some are known and used as toxins. As these materials become more readily accessible for legitimate and illegitimate purposes, the need to have a standardized method to isolate and identify pharmacologically and toxicologically active compounds, present in these plant materials, has become acute. Approaches to this problem have been developed on the individual genus/species level as well as through more thorough approaches that analyze a variety of toxins in tissue and blood samples. Here we show a standardized approach to the extraction and identification of toxins in several different plant species containing different alkaloids. Since we expect to see species-to-species variation in level of active compounds in a particular genus, the method must be ubiquitous and robust enough to translate to other species of the same genus. For example, the minimum protocol set should be able to identify the different toxins in both digitalis purpurea vs. digitalis lanata. This method, in particular, focuses on seed materials as they are most easily stored and transported. The universal protocol homogenizes the seed material via a freezer mill in liquid nitrogen. The homogenized material is then sequentially extracted by room temperature maceration in methanol and the resulting extract is purified by solid phase extraction, leaving the analytes in an acceptable solvent for liquid chromatography-mass spectrometry (LC-MS) analysis. The toxins can then be identified by their retention times, mass-to-charge ratio, as well as their fragmentation patterns. The species initially analyzed include aconitum napellus, atropa belladonna, cerbera odollam, conium maculatum, digitalis purpurea, strychnos nux-vomica, and nerium oleander. Each of these species contain several active compounds of interest with varying toxicities.

Universal approach to the screening of plant-based toxins from a variety of genus and species

2019 Spring National Meeting

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