Method for identification and quantification of intact teduglutide peptide using (RP)UHPLC-UV-(HESI/ORBITRAP)MS

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis

Background:

• Teduglutide is a glucagon-like peptide 2 analogue used to treat short bowel syndrome.
• Analyzing protein-based therapeutics like teduglutide presents significant challenges due to their complex structures.

Purpose of the Study:

• To develop and validate a rapid Reversed-Phase Ultra-High-Performance Liquid Chromatography-Ultraviolet-Mass Spectrometry (RP-UHPLC-UV-MS) method for teduglutide.
• To simultaneously identify, structurally characterize, and quantify intact teduglutide.

Main Methods:

• A novel RP-UHPLC-UV-(HESI/ORBITRAP)MS method was established and validated per ICH guidelines.
• Method validation included linearity, LOD/LOQ, accuracy, precision, trueness, robustness, system suitability, and specificity.
• Forced degradation studies (thermal, mechanical, light) were conducted to assess specificity and impact of mishandling.

Main Results:

• The developed method demonstrated excellent validation performance for both UV and MS signals.
• Teduglutide was identified by its exact mass, confirming the absence of post-translational modifications.
• The method successfully detected and characterized degradation/modification products, indicating stability.

Conclusions:

• This study presents the first method for simultaneous identification, structural characterization, and quantification of therapeutic teduglutide.
• The validated UHPLC-MS method is suitable for quality control and stability assessment of teduglutide formulations.
• The method's ability to detect degradation products ensures the integrity and efficacy of teduglutide treatment.