Anion exchange stationary phases based on a polyalkylpolyamine polymer layer

Abstract

An anion exchange for separating a plurality of carbohydrates includes a negatively charged substrate particle. A base polymer layer includes a first plurality of quaternary amines. The polyalkylpolyamine polymer layer is covalently attached to the base condensation polymer layer. The polyalkylpolyamine polymer layer includes a polymeric branch structure that includes a second plurality of quaternary amines. A density of the second plurality of quaternary amines increases in a direction away from the base condensation polymer layer. The anion exchange stationary phase does not have a hydroxy group spaced apart from any one of the first or the second plurality of quaternary amines by an ethyl group.

What is claimed is: 
     
         1 . An anion exchange stationary phase for separating a plurality of carbohydrates comprises:
 a) a negatively charged substrate particle;   b) a base condensation polymer layer attached to the negatively charged substrate particle, the base condensation polymer layer comprises a first plurality of quaternary amines, in which the first plurality of quaternary amines are spaced apart by either a first spacer or a second spacer, in which the base condensation polymer layer does not have a hydroxy group spaced apart from one of the first plurality of quaternary amines by an ethyl group;   c) a polyalkylpolyamine condensation polymer layer covalently attached to the base condensation polymer layer, the polyalkylpolyamine condensation polymer layer comprises a polymeric branch structure, the polymeric branch structure includes a second plurality of quaternary amines, in which the second plurality of quaternary amines are spaced apart by the first spacer or the second spacer, in which a density of the second plurality of quaternary amines increases in a direction away from the base condensation polymer layer, in which the polyalkylpolyamine condensation polymer layer does not have a hydroxy group spaced apart from one of the second plurality of quaternary amines by an ethyl group.   
     
     
         2 . The anion exchange stationary phase of  claim 1 , in which the negatively charged substrate particle comprises a crosslinked divinylbenzene and ethylvinyl benzene copolymer, in which at least a surface of the negatively charged substrate particle includes sulfonate groups. 
     
     
         3 . The anion exchange stationary phase of  claim 1 , in which the negatively charged substrate particle comprises a crosslinked divinylbenzene and ethylvinyl benzene copolymer, in which at least a surface of the negatively charged substrate particle includes carboxylate groups. 
     
     
         4 . The anion exchange stationary phase of  claim 1 , in which the base condensation polymer layer is positively charged and ionically attached to the negatively charged substrate particle. 
     
     
         5 . The anion exchange stationary phase of  claim 1 , in which the first spacer comprises a chemical formula of (—CH 2 —) x  and the second spacer comprises a chemical formula of (—CH 2 —) y , where x and y each independently range from 3 to 6. 
     
     
         6 . The anion exchange stationary phase of  claim 1 , in which the first spacer comprises a first alkyl and the second spacer comprises a second alkyl, in which the first alkyl and the second alkyl are both a linear and unsubstituted alkyl. 
     
     
         7 . The anion exchange stationary phase of  claim 1 , in which the first spacer comprises a linear and unsubstituted alkyl and the second spacer comprises an arylalkyl. 
     
     
         8 . The anion exchange stationary phase of  claim 1 , in which the first spacer is selected from group consisting of an alkyl, a dialkylether, a cycloalkyl, an arylalkyl, and a combination thereof. 
     
     
         9 . The anion exchange stationary phase of  claim 1 , in which the first spacer comprises an alkyl and the second spacer is selected from group consisting of an alkyl, an arylalkyl, and a combination thereof. 
     
     
         10 . An anion exchange stationary phase for separating a plurality of carbohydrates, the anion exchange stationary phase formed by a method comprising:
 reacting a polyhalohydrocarbon with a polyalkylpolyamine to form a base condensation polymer layer on a negatively charged substrate particle; and   reacting the base condensation polymer layer with a number of reaction cycles to form a polyalkylpolyamine condensation polymer layer, in which the number of reaction cycles ranges from about three to about ten and each reaction cycle includes a polyhalohydrocarbon treatment and a polyalkylpolyamine treatment.   
     
     
         11 . The anion exchange stationary phase of  claim 10  further comprising:
 reacting the polyalkylpolyamine condensation polymer layer with a monohaloalkane treatment. 
 
     
     
         12 . The anion exchange stationary phase of  claim 10 , in which the negatively charged substrate particles are contained as a packed bed in a column, in which the reacting of the polyhalohydrocarbon with the polyalkylpolyamine comprises: flowing a solution of the polyhalohydrocarbon and the polyalkylpolyamine through the column to form the base condensation polymer layer on the negatively charged substrate particles. 
     
     
         13 . The anion exchange stationary phase of  claim 12 , in which the polyhalohydrocarbon treatment comprises: flowing a solution of the polyhalohydrocarbon through the column; the polyalkylpolyamine treatment comprises: flowing a solution of the polyalkylpolyamine through the column; and the monohaloalkane treatment comprises: flowing a solution of the monohaloalkane through the column. 
     
     
         14 . The anion exchange stationary phase of  claim 10 , in which the number of reaction cycles ranges from about 3 to about 4. 
     
     
         15 . The anion exchange stationary phase of  claim 10 , in which the negatively charged substrate particle comprises a crosslinked divinylbenzene and ethylvinyl benzene copolymer, in which at least a surface of the negatively charged substrate particle includes sulfonate groups. 
     
     
         16 . The anion exchange stationary phase of  claim 10 , in which the negatively charged substrate particle comprises a crosslinked divinylbenzene and ethylvinyl benzene copolymer, in which at least a surface of the negatively charged substrate particle includes carboxylate groups. 
     
     
         17 . The anion exchange stationary phase of  claim 10 , in which the polyhalohydrocarbon comprises a material selected from group consisting of a dihaloalkane, a dihalodialkylether, a dihalocycloalkane, a trihaloarylalkane, and a combination thereof. 
     
     
         18 . The anion exchange stationary phase of  claim 10 , in which the polyalkylpolyamine comprises a material selected from group consisting of a polyalkyltriamine, a polyalkyldiamine, and a combination thereof. 
     
     
         19 . The anion exchange stationary phase of  claim 10 , in which all amines of the polyalkylpolyamine are tertiary amines. 
     
     
         20 . The anion exchange stationary phase of  claim 10 , in which the polyhalohydrocarbon is selected from the group consisting of a dibromobutane, a dibromopentane, a dibromohexane, a tribromomethylbenzene, and a combination thereof. 
     
     
         21 . The anion exchange stationary phase of  claim 10 , in which the polyalkylpolyamine is selected from the group consisting of a pentamethyldipropyltriamine, a pentamethyldihexylltriamine, a permethylated spermine, a permethylated spermidine, and a combination thereof. 
     
     
         22 . The anion exchange stationary phase of  claim 10 , in which the polyhalohydrocarbon is dibromobutane and the polyalkylpolyamine is pentamethyldihexylltriamine. 
     
     
         23 . The anion exchange stationary phase of  claim 10 , in which the polyhalohydrocarbon is a trihaloalkylaryl and the polyalkylpolyamine is an alkyldiamine. 
     
     
         24 . The anion exchange stationary phase of  claim 10 , in which the polyhalohydrocarbon is tribromomethylbenzene and the polyalkylpolyamine is tetramethylhexanediamine. 
     
     
         25 . An anion exchange stationary phase for separating a plurality of carbohydrates, the anion exchange stationary phase comprises:
 A) a negatively charged substrate particle;   B) a base condensation polymer layer attached to the negatively charged substrate particle, the base condensation polymer layer comprises a reaction product of
 i) a first polyhalohydrocarbon, and 
 ii) a first polyalkylpolyamine; 
   C) a first alkyl condensation reaction product covalently attached to the base condensation polymer layer, the first alkyl condensation reaction product comprises a reaction product of
 i) an amine group of the base condensation polymer layer, and 
 ii) a second polyhalohydrocarbon, in which the amine group of the base condensation polymer layer includes a positive charge so that the base condensation polymer layer is ionically coupled to the negatively charged substrate particle; 
   D) a first polyalkylpolyamine condensation reaction product covalently attached to the first alkyl condensation reaction product, the first polyalkylpolyamine condensation reaction product comprises a reaction product of
 i) a halide group of the second polyhalohydrocarbon, and 
 ii) a second polyalkylpolyamine; and 
   E) a second alkyl condensation reaction product covalently attached to the first polyalkylpolyamine condensation reaction product, the second alkyl condensation reaction product comprises a reaction product of
 i) an amine group of the first polyalkylpolyamine condensation reaction product, and 
 ii) a third polyhalohydrocarbon; 
   F) a second polyalkylpolyamine CRP covalently attached to the second alkyl CRP, the second polyalkylpolyamine CRP comprises a reaction product of
 i) a halide group of the third polyhalohydrocarbon and 
 ii) a third polyalkylpolyamine; 
   G) a third alkyl CRP covalently attached to the second polyalkylpolyamine CRP, the third alkyl CRP includes a reaction product of
 i) an amine group of the second polyalkylpolyamine CRP, and 
 ii) a fourth polyhalohydrocarbon; and 
   H) a third polyalkylpolyamine CRP is covalently attached to the third alkyl CRP, the third polyalkylpolyamine CRP includes a reaction product of
 i) a halide group of the fourth polyhalohydrocarbon, and 
 ii) a fourth polyalkylpolyamine. 
   
     
     
         26 . The anion exchange stationary phase of  claim 25 , in which the first polyhalohydrocarbon, second polyhalohydrocarbon, third polyhalohydrocarbon, and fourth polyhalohydrocarbon comprise a dihaloalkane; and the first polyalkylpolyamine, second polyalkylpolyamine, third polyalkylpolyamine, and fourth polyalkylpolyamine comprise a polyalkyltriamine. 
     
     
         27 . A method of using an anion exchange stationary phase for separating a plurality of carbohydrates in a sample, the anion exchange stationary phase comprising:
 a) a negatively charged substrate particle;   b) a base condensation polymer layer attached to the negatively charged substrate particle, the base condensation polymer layer comprises a first plurality of quaternary amines, in which the first plurality of quaternary amines are spaced apart by either a first spacer or a second spacer, in which the base condensation polymer layer does not have a hydroxy group spaced apart from one of the first plurality of quaternary amines by an ethyl group;   c) a polyalkylpolyamine condensation polymer layer covalently attached to the base condensation polymer layer, the polyalkylpolyamine condensation polymer layer comprises a polymeric branch structure, the polymeric branch structure includes a second plurality of quaternary amines, in which the second plurality of quaternary amines are spaced apart by the first spacer or the second spacer, in which a density of the second plurality of quaternary amines increases in a direction away from the base condensation polymer layer, in which the polyalkylpolyamine condensation polymer layer does not have a hydroxy group spaced apart from one of the second plurality of quaternary amines by an ethyl group, the method comprising:
 flowing an eluent through a chromatography column, the chromatography column containing the anion exchange stationary phase, in which the eluent includes a hydroxide; 
 injecting the sample comprising a plurality of carbohydrates into the chromatography column; 
 separating at least one carbohydrate from the sample injected into the chromatography column; and 
 detecting the at least one carbohydrate at a detector. 
   
     
     
         28 . The method of  claim 27 , in which the at least one carbohydrate is a branched glycan.