High-Efficiency Liquid Chromatographic Separation Utilizing Long Monolithic Silica Capillary Columns

• Published in: Analytical chemistry (Washington) Vol. 80; no. 22; pp. 8741 - 8750
• Main Authors: Miyamoto, Kosuke, Hara, Takeshi, Kobayashi, Hiroshi, Morisaka, Hironobu, Tokuda, Daisuke, Horie, Kanta, Koduki, Kodai, Makino, Satoshi, Núñez, Oscar, Yang, Chun, Kawabe, Takefumi, Ikegami, Tohru, Takubo, Hirotaka, Ishihama, Yasushi, Tanaka, Nobuo
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 15-11-2008
• Subjects: Analytical chemistry; Animals; Cattle; Chemical reactions; Chemistry; Chromatographic methods and physical methods associated with chromatography; Chromatography; Chromatography, High Pressure Liquid - methods; Exact sciences and technology; Hydrocarbons; Hydrocarbons, Aromatic - isolation & purification; Other chromatographic methods; Peptides; Peptides - isolation & purification; Peptides - metabolism; Proteins; Silica; Silicon Dioxide; Capillary column; Water; Performance evaluation; Hydrocarbon; Peptides; Benzene derivatives; HPLC chromatography; Retention factor; Liquid chromatography; Permeability; Silica; Protein; Reversed phase chromatography; Deadtime; Efficiency; Acetonitrile; Height equivalent to a theoretical plate
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac801042c

Long monolithic silica-C18 capillary columns of 100 μm i.d. were prepared, and the efficiency was examined using reversed-phase HPLC under a pressure of up to 47 MPa. At linear velocities of 1−2 mm/s, 100000−500000 theoretical plates could be generated with a single column (90−440 cm in length) using an acetonitrile−water (80/20) mobile phase with a column dead time (t 0) of 5−40 min. It was possible to prepare columns with a minimum plate height of 8.5 ± 0.5 μm and permeability of (1.45 ± 0.09) × 10−13 m2. The chromatographic performance of a long octadecylsilylated monolithic silica capillary column was demonstrated by the high-efficiency separations of aromatic hydrocarbons, benzene derivatives, and a protein digest. The efficiency for a peptide was maintained for an injection of up to 0.5−2 ng. When three 100 μm i.d. columns were connected to form a 1130−1240 cm column system, 1000000 theoretical plates were generated for aromatic hydrocarbons with retention factors of up to 2.4 with a t 0 of 150 min. The fact that very high efficiencies were obtained for the retained solutes suggests the practical utility of these long monolithic silica capillary columns.