Analysis of psilocybin has been somewhat overlooked through history because of the controlled nature of the substance. We adapted literature methods (Table ) for our equipment and found most failed to reproducibly produce a single well-resolved peak for the compound. Many of these methods employed typical C18 or C8 columns, readily available eluents, and typical temperature programming since these are important criteria for robust HPLC analysis. We found the main source of inaccuracies with certain literature methods were related to either high concentrations of eluent additive employed or unbuffered formic acid use. Psilocybin and psilocin remain infrequently analyzed and standard analytical methods are not well developed or validated. Further, research is partially limited to academic laboratories with special clearance and advanced research equipment. The handful of analytical methods developed in academic settings tend to be more selective and potentially more accurate; however, they are also costly and inherently less available to the average analyst. Other quantitative HPLC methods provided sound characterization at the expense of acquisition time. The objective of this research was to achieve a reliable peak shape and maximize performance for the quantitation of psilocybin and psilocin. We aimed to keep the method short for use as a high-throughput purity check. Here, we present our results and discuss HPLC method development along with the pitfalls of commonly used HPLC eluent additives for this analysis.