Metabolite identification using an ion mobility enhanced data-independent acquisition strategy and automated data...

Area of Science:

• Analytical Chemistry
• Mass Spectrometry
• Pharmacokinetics

Background:

• Liquid chromatography-mass spectrometry is crucial for drug metabolism and pharmacokinetics studies.
• Data-independent acquisition (MS E) aids metabolite detection but lacks quadrupole-resolved spectra.
• Ion mobility separation, integrated into High-Definition MS E (HDMS E), aims to improve metabolite resolution and characterization efficiency.

Purpose of the Study:

• To compare the efficacy of High-Definition Data-Dependent Acquisition (HD-DDA), MS E, and HDMS E methods.
• To evaluate automated software processing using Mass-MetaSite and WebMetabase for metabolite identification.
• To assess the impact of ion mobility on the accuracy and confidence of metabolite structure elucidation.

Main Methods:

• Metabolite identification was performed on incubations of glucagon-like peptide-1 (7-37) and verapamil hydrochloride.
• Experiments utilized UPLC, a VION IMS QTOF mass spectrometer under UNIFI control.
• Data processing involved MassMetaSite and WebMetabase for peak detection and spectral interpretation.

Main Results:

• HDMS E and MS E proposed identical structures for verapamil and GLP-1 metabolites.
• HDMS E demonstrated a slightly higher ratio of structurally matched to mismatched product ions compared to MS E and HD-DDA.
• Ion mobility acquisition in HDMS E enhanced confidence in proposed metabolite structures.

Conclusions:

• HDMS E is an effective technique for elucidating metabolite structures in both small molecules and peptides.
• The approach offers excellent accuracy and quality with minimal compound-specific optimization.
• HDMS E provides reliable metabolite characterization, improving upon traditional MS E methods.