Packing material for HILIC columns, HILIC column filled with same, and method for analyzing oligosaccharide with use of same
Provided is a packing material for HILIC columns for more accurately and more easily performing oligosaccharide analysis by liquid chromatography; an HILIC column which is filled with the packing material for HILIC columns; and a method for analyzing an oligosaccharide with use of this packing material for HILIC columns A packing material for HILIC columns according to the present invention is composed of particles, each of which is obtained by reacting glycidol to a hydroxyl group of a porous cross-linked polymer base material having the hydroxyl group, and which have a hydrophilicity index of 2.30 or more and a surface-pH index of from 0.95 to 1.05.
1. A packing material for HILIC columns, comprising a cross-linked polymer base material having a hydroxyl group,
wherein the cross-linked polymer base material is porous particles;
a glycidol is reacted with the hydroxyl group of the cross-linked polymer base material;
the packing material has a hydrophilicity index of 2.30 or more; and
the packing material has a surface-pH index of from 0.95 to 1.05,
wherein the hydrophilicity index is defined by a separation coefficient of α1 (U/2dU), when performing a liquid chromatography measurement of a uridine (U) and a 2′-deoxyuridine (2dU) in a HILIC separation mode condition;
the surface-pH index is a separation coefficient of α2 (Tb/Tp), when performing a liquid chromatography measurement of a theobromine (Tb) and a theophylline (Tp) in a HILIC separation mode condition;
the separation coefficient α1 (U/2dU) and the separation coefficient α2 (Tb/Tp) are ratios of retention factors k of the materials, respectively;
each retention factor k is defined by k=(t−t 0 )/t 0 , wherein t 0 is an elution time of toluene and, t is an elution time of each substance; and
a condition of the liquid chromatography measurement in the HILIC separation mode is shown as below:
a column temperature is 30° C.; and
a mobile phase is a mixed liquid of acetonitrile and an aqueous solution in a ratio (acetonitrile/aqueous solution) of 90/10 (volume before mixing) wherein the aqueous solution contains 10 mM acetic acid and 10 mM ammonium acetate.
2. The packing material for HILIC columns according to claim 1 , wherein the porous cross-linked polymer base material having a hydroxyl group is a copolymer of vinyl alcohol and triallyl isocyanurate.
3. A HILIC column, comprising
the packing material for HILIC columns according to claim 1 , and
a liquid chromatography casing in which the packing material for HILIC columns is filled.
4. A method for analyzing oligosaccharides, comprising
analyzing oligosaccharides in a HILIC mode using the HILIC column according to claim 3 .
5. The method for analyzing oligosaccharides according to claim 4 ,
wherein a composition of a mobile phase is constant during the measurement (in an isocratic elution condition) and a column temperature is constant during the measurement method for analyzing oligosaccharides.
6. The method for analyzing oligosaccharides according to claim 4 ,
wherein a molecular size of the oligosaccharides to be analyzed is 17 mer or more.
7. The packing material for HILIC columns according to claim 1 ,
wherein a particle size of the porous particles is 1 to 30 μm as a volume average particle diameter, and
an average pore diameter of pores forming the porous particles is from 3 to 30 nm.