Enhanced sequence coverage in tryptic fragment analysis by two-dimensional HPLC/MS using a monolithic silica capillary column

• Published in: Bioscience, biotechnology, and biochemistry Vol. 70; no. 9; pp. 2154 - 2159
• Main Authors: Morisaka, H.(Kyoto Univ. (Japan)), Hata, K, Mima, J, Tanigawa, T, Furuno, M, Ishizuka, N, Tanaka, N, Ueda, M
• Format: Journal Article
• Language: English
• Published: Tokyo Japan Society for Bioscience, Biotechnology, and Agrochemistry 01.09.2006; Japan Society for Bioscience Biotechnology and Agrochemistry; Oxford University Press
• Subjects: Amino Acid Sequence; AMINO ACID SEQUENCES; Biological and medical sciences; capillary column; CHROMATOGRAPHIE; CHROMATOGRAPHY; Chromatography, High Pressure Liquid - methods; Chromatography, Ion Exchange; CROMATOGRAFIA; DESEMPENO DE LA MAQUINARIA; EQUIPMENT PERFORMANCE; ESPECTROMETRIA DE MASAS; Fundamental and applied biological sciences. Psychology; HPLC; LC/MS; MASS SPECTROMETRY; Molecular Sequence Data; monolithic column; multi-dimensional HPLC; PEPTIDE; PEPTIDES; PEPTIDOS; PERFORMANCE DU MATERIEL; Proteomics - methods; SECUENCIAS DE AMINOACIDOS; Sequence Analysis, Protein - methods; SEQUENCE D'ACIDES AMINES; Serum Albumin, Bovine - chemistry; Serum Albumin, Bovine - metabolism; SPECTROMETRIE DE MASSE; Spectrometry, Mass, Electrospray Ionization - methods; Trypsin - metabolism; LC/MS; capillary column; Monolithic column; HPLC chromatography; multi-dimensional HPLC; Fragment; Silica
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.60115

The HPLC/MS system, in which a monolithic silica capillary column is directly connected to an electronspray-ionization mass spectrometer, showed superior performance at high mobile phase linear velocity. A two-dimensional (2D) HPLC/MS system was established, using an ion-exchange particle-packed capillary column at the first dimension and a monolithic silica capillary column at the second dimension. In an analysis of tryptic fragments from bovine serum albumin, an 81% sequence coverage, obtained by the 2D-HPLC/MS system, increased by 23% as compared to a ID-HPLC/MS system. This 2D-HPLC/MS system using a monolithic silica capillary column should be useful for enhancing sequence coverage of tryptic fragments in proteomics.