RT-Pred: A web server for accurate, customized liquid chromatography retention time prediction of chemicals

Abstract

High-performance liquid chromatography (HPLC) together with mass spectrometry (MS) is routinely used to separate, identify and quantify chemicals. HPLC data also provides retention time (RT) which can be aligned with structural data. Recent developments in machine learning (ML) have improved our ability to predict RTs from known or postulated chemical structures, allowing RT data to be used more effectively in LC-MS-based compound identification. However, RT data is highly specific to each chromatographic method (CM) and hundreds of different CMs with interdependent parameters are used in separations. This has limited the application of ML-based RT predictions in compound identification. Here we introduce an easy-to-use RT prediction webserver (called RT-Pred) that predicts RTs for molecules across most chromatographic setups. RT-Pred not only supports its own in-house CM-specific RT predictors, it allows users to easily train a custom RT-Pred model using their own RT data on their own CM and to predict RTs with that custom model. RT-Pred also supports RT and compound searches against its own database of millions of predicted RTs spanning >40 different CMs. RT-Pred is also uniquely capable of accurately identifying compounds that will elute in the void volume or be retained on the column. Including this void/retained/eluted classifier significantly improves RT-Pred's performance. Tests indicate that RT-Pred had an average coefficient of determination (R²) of 0.95 over 20 different CMs. Comparisons of RT-Pred against other RT predictors showed that RT-Pred achieved lower mean absolute errors and higher R² scores than any other published RT predictor. RT-Pred is freely available at https://rtpred.ca.