Long-Term System Suitability Evaluation for Mass Accuracy in the Analysis of Small Molecules by High-Resolution Mass Spectrometry

Abstract

High-resolution mass spectrometry (HRMS) is critical for the identification and characterization of (un)known organic chemicals. In this regard, ensuring high mass accuracy in HRMS instruments is essential for reliable results in nontarget and suspect screening. This study presents a practical approach for evaluating and maintaining mass accuracy over time using ultrahigh pressure liquid chromatography coupled with electrospray ionization Orbitrap HRMS. A set of 13 reference standards, encompassing a range of polarities and chemical families, was analyzed before and after sample analysis batches to assess the impact of various factors on the instrumental performance regarding mass accuracy. The aim is not to recalibrate the system but to provide a reliable snapshot of the mass accuracy over time. The study found that the positive ionization mode exhibited higher accuracy and precision compared with the negative mode. Factors affecting mass accuracy included calibration quality, the number of batch injections, and the time between calibrations, where the two latter factors were related to each other. Results suggest that performing system suitability tests for high-resolution accurate masses with two injections before and after sample analysis is adequate for ensuring acceptable mass spectrometric performance for robust and reliable HRMS data acquisition, but performing three injections is recommended. This protocol ensures that informed decisions can be made with regard to the mass accuracy, the calibration, and a potential recalibration before HRMS data acquisition is performed.