PeakClimber: A software tool for the accurate quantification of complex multianalyte HPLC chromatograms using the exponential Gaussian function

High-performance liquid chromatography (HPLC) is a common medium-throughput technique to quantify the components of often complex mixtures like those typically obtained from biological tissue extracts. However, analysis of HPLC data from complex multianalyte samples is hampered by a lack of tools to accurately determine the precise analyte quantities on a level of precision equivalent to mass-spectrometry approaches. To address this problem, we developed a tool we call PeakClimber, that uses a sum of exponential Gaussian functions to accurately deconvolve overlapping, multianalyte peaks in HPLC traces. Here we analytically show that HPLC peaks are well-fit by an exponential Gaussian function, that PeakClimber more accurately quantifies known peak areas than standard industry software for both HPLC and mass spectrometry applications, and that PeakClimber accurately quantifies differences in triglyceride abundances between colonized and germ-free fruit flies.