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Analysis of calcitonin and its analogues by capillary zone electrophoresis and matrix‐assisted laser‐desorption ionization time‐of‐flight mass spectrometry
Author(s) -
Amini Ahmad,
Olofsson IngMarie
Publication year - 2004
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200401722
Subject(s) - chemistry , capillary electrophoresis , mass spectrometry , chromatography , desorption , cationic polymerization , analyte , analytical chemistry (journal) , electrophoresis , capillary electrophoresis–mass spectrometry , matrix assisted laser desorption/ionization , capillary action , time of flight mass spectrometry , ionization , matrix (chemical analysis) , ion , electrospray ionization , adsorption , materials science , polymer chemistry , organic chemistry , composite material
Capillary zone electrophoretic (CZE) separations and mass spectrometric analysis of salmon calcitonin and related analogues were performed to generate electrophoresis and mass fingerprints for quality control of the recombinant polypeptide pharmaceutical salmon calcitonin. The calcitonins and their corresponding tryptic digests were successfully separated by CZE at low pH in fused silica capillaries dynamically modified with poly‐cationic polymers. The poly‐cationic modified inner surface of the fused silica capillaries generated a strong anionic electroosmotic flow (EOF). Analytes of negative, neutral, and positive charge were all swept through the capillary toward the positive electrode. Compared to Polybrene‐coated capillaries, capillaries coated with PEI showed a markedly slower but much more stable electroosmotic flow. The migration order of the analytes was predicted by comparing approximate values of the charge to (molecular mass) 2/3 ratios. The predicted migration order was confirmed by off‐line analysis of CZE fractions with matrix‐assisted laser‐desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS).