As GLP-1-based therapeutics and other complex peptides continue to advance, bioanalytical methods must balance sensitivity, selectivity, quantitative performance and development speed. Semaglutide, tirzepatide and retatrutide present analytical challenges that can require careful optimization of chromatography and mass spectrometry conditions. 

At SCIEX Summit 2026, Veloxity Labs President and CEO Shane Needham, PhD, presented a head-to-head evaluation of high-resolution mass spectrometry and triple quad LC-MS/MS workflows for quantitative peptide bioanalysis.

Using a single sample preparation and LC-MS/MS analysis in human plasma, the study compared the SCIEX 6500+ triple quad system with the SCIEX 8600 ZenoTOF HRMS system for semaglutide, tirzepatide and retatrutide.

Key findings

Both workflows achieved a 0.5 ng/mL LLOQ for all three peptides and maintained a linear dynamic range of 0.5–1,000 ng/mL. The HRMS workflow achieved this performance using 5 µL injection volume compared with 15 µL for the triple quad workflow.

But quantitative performance told only part of the story.

Comprehensive MS1/MS2 data from the HRMS workflow provided additional analytical information for charge-state assessment, parent-mass confirmation and flexible product-ion selection. The ability to evaluate multiple product ions from the same acquisition and combine informative fragment signals also reduced the need for iterative optimization.

In this evaluation, flexible product-ion selection and fragment summing supported approximately 3–4X faster method development, while maintaining sensitivity across all three peptide assays.

The findings demonstrate how HRMS can complement established triple quad workflows for complex peptide bioanalysis, providing comparable quantitative sensitivity while adding selectivity, analytical insight and method-development flexibility. 

Presented by: Shane Needham, PhD, President & CEO, Veloxity Labs

Event: SCIEX Summit 2026

Date: October 7, 2026