MAD HATTER Correctly Annotates 98% of Small Molecule Tandem Mass Spectra Searching in PubChem

Metabolites provide a direct functional signature of cellular state. Untargeted metabolomics usually relies on mass spectrometry, a technology capable of detecting thousands of compounds in a biological sample. Metabolite annotation is executed using tandem mass spectrometry. Spectral library search is far from comprehensive, and numerous compounds remain unannotated. So-called in silico methods allow us to overcome the restrictions of spectral libraries, by searching in much larger molecular structure databases. Yet, after more than a decade of method development, in silico methods still do not reach the correct annotation rates that users would wish for. Here, we present a novel computational method called Mad Hatter for this task. Mad Hatter combines CSI:FingerID results with information from the searched structure database via a metascore. Compound information includes the melting point, and the number of words in the compound description starting with the letter 'u'. We then show that Mad Hatter reaches a stunning 97.6% correct annotations when searching PubChem, one of the largest and most comprehensive molecular structure databases. Unfortunately, Mad Hatter is not a real method. Rather, we developed Mad Hatter solely for the purpose of demonstrating common issues in computational method development and evaluation. We explain what evaluation glitches were necessary for Mad Hatter to reach this annotation level, what is wrong with similar metascores in general, and why metascores may screw up not only method evaluations but also the analysis of biological experiments. This paper may serve as an example of problems in the development and evaluation of machine learning models for metabolite annotation.

Automated summary

Metabolites reflect the cellular state and can be analyzed using mass spectrometry. However, the current annotation methods in metabolomics are not achieving the desired annotation rates. In this paper, a novel computational method called Mad Hatter is presented, which combines results from CSI:FingerID and a structure database to improve annotation accuracy.