Improvement of separation efficiencies of anion-exchange chromatography using monolithic silica capillary columns modified with polyacrylates and polymethacrylates containing tertiary amino or quaternary ammonium groups.
Anion-exchange (AEX) columns were prepared by on-column polymerization of acrylates and methacrylates containing tertiary amino or quaternary ammonium groups on monolithic silica in a fused silica capillary modified with anchor groups. The columns provided a plate height (H) of less than 10 microm at optimum linear velocity (u) with keeping their high permeability (K=9-12 x 10(-14) m2). Among seven kinds of AEX columns, a monolithic silica column modified with poly(2-hydroxy-3-(4-methylpiperazin-1-yl)propyl methacrylates) (HMPMA) showed larger retentions and better selectivities for nucleotides and inorganic anions than the others. The HMPMA column of 410 mm length produced 42,000-55,000 theoretical plates (N) at a linear velocity of 0.97 mm/s with a backpressure of 3.8 MPa. The same column could be employed for a fast separation of inorganic anions in 1.8 min at a linear velocity of 5.3 mm/s with a backpressure of 20 MPa. In terms of van Deemter plot and separation impedance, the HMPMA column showed higher performance than a conventional particle-packed AEX column. The HMPMA column showed good recovery of a protein, trypsin inhibitor, and it was applied to the separation of proteins and tryptic digest of bovine serum albumin (BSA) in a gradient elution, to provide better separation compared to a conventional particle-packed AEX column.
Watanabe
Yuta
Y
Department of Biomolecular Engineering, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan.
Ikegami
Tohru
T
Horie
Kanta
K
Hara
Takeshi
T
Jaafar
Jafariah
J
Tanaka
Nobuo
N
D000180:Acrylic Resins / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D002852:Chromatography, Ion Exchange / Q000295:instrumentation* / Q000379:methods*; D011109:Polymethacrylic Acids / Q000737:chemistry*; D011506:Proteins / Q000032:analysis / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000032:analysis* / Q000378:metabolism; D012822:Silicon Dioxide / Q000737:chemistry*
D000180:Acrylic Resins; D011109:Polymethacrylic Acids; D011506:Proteins; C030613:polymethacrylic acid; D012710:Serum Albumin, Bovine; C006903:carbopol 940; D012822:Silicon Dioxide