Developing A Method To Extract Alkaloids From The Secretions Of Salamanders Found In Northwestern North Carolina

Many species of salamanders secrete noxious chemicals as a defense mechanism. Two salamanders found in western North Carolina, P. yonahlossee and P. montanous, belong to the same genus, but show vastly different coloring. P. yonahlossee displays a bright red dorsal patch, while P. montanous is uniformly grey. Recent research conducted in the Biology Department at Appalachian State University using clay models to represent salamanders showed that predatory birds avoided models with bright red spots, suggesting that the defense mechanism was effective. It is not known, however, if the P. yonahlossee secrets noxious substances, or if the red patch is simply a form of mimicry. The goal of this work is to determine if the P. yonahlossee secrets noxious substances. Recent research conducted on poisonous frogs suggests that the noxious compounds in salamanders could be alkaloids. I used nicotine as a model alkaloid and compared the ability of liquid-liquid extraction (LLE) and solid phase extraction (SPE) to recover alkaloids from salamander secretion. GC/MS was used to quantify all extracts. Although previous students optimized the LLE method, we have been unable to reproduce their results and have done experiments to determine if nicotine is lost in the evaporation step of the extraction. We used two different GCs and determined that one of them provides cleaner chromatograms, probably because it has fewer users. Although our samples have aged in the freezer and our proposed method is only semi-quantitative, this research has the potential to answer important questions about salamanders in western North Carolina that can be applied to other regions. In addition, to our knowledge, no other methods in the literature have used SPE to recover alkaloids from salamander secretion, so there is potential to contribute a new, robust method.