Metabolite Identification

Characterize and quantify metabolites with speed, efficiency, and confidence.

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Using Ion Mobility LC-MS and LC-MS/MS to Resolve and Identify Isobaric Glucuronide Metabolites

Common metabolic pathways often give rise to multiple instances of isobaric metabolites such as, (but not limited to), multiple mono-hydroxylations and di-hydroxylations with and without phase II conjugations, such as glucuronidation. LC-MS analysis of metabolic pathways for drugs requires accurate detection and identification of closely eluting species with similar RT’s and spectral profiles. This is further complicated by shifting abundance profiles at different time points, species differences and RT shifts due to biological matrix effects. Modern chromatography (and mass spectrometry) is sufficient in many cases, however isobaric species remain problematic. Ensuring closely eluting species have not shifted by RT requires standard addition or commixing of matrix and true isobaric coelutions may go undetected with conventional LC-MS approaches as they rely solely on discrimination using precursor mass. Furthermore, the MS/MS patterns from related compounds are often extremely similar and coelutions give rise to complex spectra which can hinder identification and structural elucidation. MS/MS of metabolites at low concentrations may also introduce noise from matrix at high concentrations. Review this poster to find out more information.

Use of Ion Mobility for DMPK Applications Throughout the Drug Development Programme of Small Molecules

Dr. Catherine Holdsworth discusses how Covance routinely utilizes ion mobility technology. She demonstrates its benefits for metabolite profiling studies, including resolving co-eluting isomeric peaks, and tracking, correlating, and comparing metabolites between different chromatographic systems.

Routine Ion Mobility and UNIFI Software as Tools for Efficient and Robust Metabolite Characterisation

Dr. Jan Boerma reviews how he and Unilabs Bioanalytical Solutions use ion mobility to reduce noise and reveal low abundance diagnostic ions, remove endogenous components, and obtain clean, high-energy spectra of co-eluting components with different drift times. He also shows how the UNIFI Scientific Information System helps to streamline metbolite ID by uncovering primary and secondary metabolites and identify unexpected metabolites.

Metabolism of Peptide Therapeutics Using UNIFI

Dr. Fabrice Bonche discusses how Sanofi utilizes HRMS and the UNIFI Scientific Information System to investigate metabolism of peptide therapeutics. He examines raw MS spectra and chromatography, considerations for "semi" quantification; and the full PK profile.

White paper: Biopharmaceutical System Solution with UNIFI Scientific Information System for the Evaluation of Peptide Catabolism

This white paper discusses the use of the Waters Biopharmaceutical Peptide Mapping solution for the characterization of peptide catabolism, using a dedicated workflow.

Data Independent and Data Dependent Acquisition Strategies Combined With Ion Mobility for Drug Metabolism and Pharmacokinetics

This poster presented at 12th Annual Great Lakes DMDG meeting from May 4-5th in Michigan.

Ion Mobility-enabled Data-dependent Experiments Distinguishing Co-eluting Isomeric Metabolites Using an IMS-QTof Mass Spectrometer

Co-eluting metabolites attributed to glucuronides of dihydroxylated metabolites were successfully characterised using IMS-enabled DDA, generating two distinct precursor ion MS spectra and product ion MS/MS spectra for the drift time separated metabolites. The m/z and drift time filtered data provide cleaner, unambiguous spectra and increases confidence in structural assignment compared with simple m/z-selective DDA.

Utilization of Ion Mobility Enabled Collisional Cross Section Measurements for the Comparison of Metabolites across Differing Chromatographic Methods

When metabolites are successfully analyzed across different chromatographic methods, it reduces the need to reanalyze samples and allows comparisons to be made across studies using both HPLC and UPLC methods. Ion mobility can play an instrumental role by discriminating isomeric metabolites based on precise measurement of their ion mobility drift times and collisional cross section areas. Covance illustrates how they detected Nefazodone metabolites across two methods. CCS confirmed which isomers were not detected.

What are Ion Mobility MS and Collision Cross Section Data?

Learn about the fundamentals of ion mobility mass spectrometry (IMS) and the benefits of collisional-cross section (CCS) data.

Resolution and Characterisation of Co-Eluting Isomeric Metabolites by Collisional Cross Section Measurements Using a Novel Geometry Travelling Wave IMS QToF Mass Spectrometer

ASMS 2016 poster by Covance Laboratories demonstrating resolution of co-eluting Nefazodone isomeric metabolites using ion mobility mass spectrometry.