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Key Advantages of LC-MS/MS High sensitivity for low-abundance peptides Detects trace-level impurities and minor variants that are critical in pharmaceutical analysis Accurate mass measurement for sequence confirmation High-resolution mass spectrometry ensures precise molecular weight determination Fragmentation analysis (MS/MS) for amino acid mapping Generates sequence-specific fragment ions (b/y ions) to confirm peptide structure Ability to detect minor modifications and impurities Identifies PTMs such as oxidation, deamidation, and fatty acid conjugation Comparison with Other Techniques Why LC-MS/MS is the Gold Standard Combines chromatographic separation (LC) with mass-based detection (MS/MS) Handles complex peptide mixtures and modified analogs effectively Meets regulatory expectations for peptide characterization Provides high reproducibility and robustness for routine analysis 3: Workflow for Peptide Sequencing of GLP-1 Peptide Using LC-MS/MS The workflow for Peptide Sequencing of GLP-1 Peptide using LC-MS/MS involves a systematic process of sample preparation, separation, ionization, mass analysis, fragmentation, and data interpretation to accurately determine peptide structure and modifications