Accelerating Method Development and Manufacturing of GLP-1 Analogs with LCUV/ MS

Significance of the Topic

Glucagon-like peptide-1 receptor agonists (GLP-1 RAs) are critical in managing type 2 diabetes and obesity. Rapid market growth for these therapeutics imposes greater demands on manufacturing and quality control workflows. Orthogonal analytical approaches that combine liquid chromatography with mass detection offer enhanced impurity screening and identity confirmation, helping manufacturers meet regulatory requirements and reduce risk.

Objectives and Study Overview

This application note aims to demonstrate a streamlined LC-UV/MS workflow for method development and in-process control of GLP-1 analogs. The study evaluates how integrating a compact mass detector with standard UV detection expedites impurity detection, supports out-of-specification (OOS) investigations, and improves overall laboratory productivity.

Used Methodology and Instrumentation

This workflow couples the ACQUITY QDa II Mass Detector and Empower 3 CDS with the Arc Premier LC system for QC-friendly operation. For high-resolution confirmation, the Xevo G3 QTof mass spectrometer and UNIFI Scientific Information System were employed. Key conditions included:

• Reversed-phase chromatography on peptide-optimized C18 columns at 60 °C
• Mobile phase of 0.1% formic acid in water and acetonitrile
• ESI positive-mode detection with full-scan acquisition from m/z 250–1500 (QDa II) and 50–2000 (QTof)
• Gradient elution and rapid tryptic digestion of exenatide to generate peptide fragments

Main Results and Discussion

Using exenatide samples from two vendors, UV detection alone indicated high purity for both. However, QDa II MS revealed a significant impurity in vendor 2 material with mass shifts of +97.1 Da. Detailed peptide mapping after tryptic digestion uncovered a unique fragment consistent with a proline insertion. Subsequent high-resolution MS confirmed this sequence variant at the C-terminus.

• LC-UV/MS enabled rapid screening for unexpected variants
• Orthogonal mass data detected impurities up to 40% abundance unseen by UV alone
• Peptide-level analysis localized the modification, avoiding extensive rework in manufacturing

Benefits and Practical Applications of the Method

The integrated workflow:

• Provides complementary UV and mass data to enhance confidence in results
• Supports quick decision-making during method development and QC release
• Reduces error rates and investigative workload in OOS scenarios
• Maintains compliance with audit-trail capability in Empower CDS

Future Trends and Applications

As peptide therapeutics diversify, compact mass detectors will become more prevalent in QC labs. Future developments may include automated spectral interpretation, real-time monitoring of bioprocess streams, and AI-driven impurity predictions. Integration with informatics platforms will further accelerate forensic analysis of product variants and process deviations.

Conclusion

The ACQUITY QDa II Mass Detector integrated with LC-UV workflows delivers critical orthogonal data for impurity detection and identity verification of GLP-1 analogs. This approach enhances lab productivity, supports regulatory compliance, and reduces the risk of undetected sequence variants.

Reference

1. Yu M. et al. Battle of GLP-1 Delivery Technologies. Advanced Drug Delivery Reviews. 2018;130:113–130. DOI:10.1016/j.addr.2018.07.009.