US 12,110,309 B2
Method for preparing analytical sample, analysis method, and kit for preparing analytical sample
Takashi Nishikaze, Kyoto (JP)
Assigned to SHIMADZU CORPORATION, Kyoto (JP)
Filed by SHIMADZU CORPORATION, Kyoto (JP)
Filed on Oct. 2, 2019, as Appl. No. 16/590,676.
Claims priority of application No. 2018-188588 (JP), filed on Oct. 3, 2018.
Prior Publication US 2020/0109159 A1, Apr. 9, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. C07H 1/00 (2006.01); G01N 33/68 (2006.01)
CPC C07H 1/00 (2013.01) [G01N 33/6848 (2013.01); G01N 2400/00 (2013.01); G01N 2440/38 (2013.01); G01N 2560/00 (2013.01)] 15 Claims
OG exemplary drawing
 
1. A method for preparing an analytical sample for analysis of a glycan contained in a sample, the method comprising:
performing an amidation reaction that amidates a lactone structure included in the glycan through contacting the sample with a reaction solution that has a pH of higher than 10;
adding an acidic solution to the reaction solution after the amidation reaction; and
purifying the sample contained in the reaction solution by using a carrier for hydrophilic interaction chromatography after the acidic solution is added to the reaction solution, wherein:
the amidation reaction amidates the lactone structure included in the glycan by aminolysis based on an interaction between an amino group and a lactone;
the reaction solution comprising at least one selected from the group consisting of ammonia, a primary amine, and a salt thereof;
wherein the concentration of ammonia, a primary amine, or a salt thereof in the reaction solution is 0.5 M or more;
a time during which the sample is in contact with the reaction solution for the amidation reaction is shorter than 30 minutes;
performing a lactonization reaction that lactonizes at least a part of sialic acids included in the glycans before the amidation reaction, wherein in the lactonization reaction, at least one selected from the group consisting of α2,3-sialic acid, α2,8-sialic acid, and α2,9-sialic acid among the sialic acids is lactonized;
an operation that reacts the sample with a dehydration condensation agent is not performed after the addition of the reaction solution to the sample, wherein: the reaction solution does not include a dehydration condensation agent that is reacted with the lactones, and wherein only contacting the sample with the amidation reaction solution is performed for the amidation reaction;
pH of the reaction solution after the acidic solution is added to the reaction solution is 10 or lower; and
the amidation reaction is performed in a liquid phase.