PeakClimber: A software tool for the accurate quantification of complex HPLC chromatograms

High-performance liquid chromatography (HPLC) is a common medium-throughput technique to quantify the components of complex mixtures like those typically obtained from biological tissue extracts. However, analysis of HPLC data from 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 bidirectional exponentially modified Gaussian (BEMG) functions to accurately deconvolve overlapping, multianalyte peaks in HPLC traces. Here we show that HPLC peaks are well-fit by a BEMG function, that PeakClimber more accurately quantifies known peak areas than standard industry software and other open-source software packages for HPLC, and that PeakClimber accurately quantifies differences in triglyceride abundances between colonized and germ-free fruit flies.