High purity chromatrographic materials comprising an ionizable modifier

Abstract

The present invention provides novel chromatographic materials, e.g., for chromatographic separations, processes for its preparation and separations devices containing the chromatographic material; separations devices, chromatographic columns and kits comprising the same; and methods for the preparation thereof. The chromatographic materials of the invention are high purity chromatographic materials comprising a chromatographic surface wherein the chromatographic surface comprises a hydrophobic surface group and one or more ionizable modifier.

1 - 17 . (canceled) 
     
     
         18 . A high purity chromatographic material comprising a chromatographic surface wherein the chromatographic surface comprises a hydrophobic surface group and one or more ionizable modifiers with the proviso that when the ionizable modifier does not contain a Zwitterion, the ionizable modifier does not contain a quaternary ammonium ion moiety;
 wherein the ionizable modifier is obtained from an ionizable modifying reagent selected from groups having the formula (III)   
       
         
           
           
               
               
           
         
         
           wherein 
           m is an integer from 1-8; 
           v is 0 or 1; 
           when v is 0, m′ is 0; 
           when v is 1, m′ is an integer from 1-8; 
           Z represents a chemically reactive group comprising at least one of: 
         
       
       
         
           
           
               
               
           
         
         
           —OH, —OR 6 , amine, alkylamine, dialkylamine, isocyanate, acyl chloride, triflate, isocyanate, thiocyanate, imidazole carbonate, NHS-ester, carboxylic acid, ester, epoxide, alkyne, alkene, azide, —Br, —Cl, or —I; 
           Y is an embedded polar functionality; 
           each occurrence of R 1  independently represents a chemically reactive group on silicon, including (but not limited to) —H, —OH, —OR 6 , dialkylamine, triflate, Br, Cl, I, vinyl, alkene, or —(CH 2 ) m″ Q; 
           each occurrence of Q is —OH, —OR 6 , amine, alkylamine, dialkylamine, isocyanate, acyl chloride, triflate, isocyanate, thiocyanate, imidazole carbonate, NHS-ester, carboxylic acid, ester, epoxide, alkyne, alkene, azide, —Br, —Cl, or —I; 
           m″ is an integer from 1-8 
           p is an integer from 1-3; 
           each occurrence of R 1′  independently represents F, C 1 -C 18  alkyl, C 2 -C 18  alkenyl, C 2 -C 18  alkynyl, C 3 -C 18  cycloalkyl, C 1 -C 18  heterocycloalkyl, C 5 -C 18  aryl, C 5 -C 18  aryloxy, or C 1 -C 18  heteroaryl, fluoroalkyl, or fluoroaryl; 
           each occurrence of R 2 , R 2′ , R 3  and R 3′  independently represents hydrogen, C 1 -C 18  alkyl, C 2 -C 18  alkenyl, C 2 -C 18  alkynyl, C 3 -C 18  cycloalkyl, C 2 -C 18  heterocycloalkyl, C 5 -C 18  aryl, C 5 -C 18  aryloxy, or C 4 -C 18  heteroaryl, —Z, or a group having the formula —Si(R′) b R″ a  or —C(R′) b R″ a ; 
           a and b each represents an integer from 0 to 3 provided that a+b=3; 
           R′ represents a C 1 -C 6  straight, cyclic or branched alkyl group; 
         
         R″ is a functionalizing group selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, cyano, amino, diol, nitro, ester, a cation or anion exchange group, an alkyl or aryl group containing an embedded polar functionality and a chiral moiety.
 each occurrence of R 6  independently represents C 1 -C 18  alkyl, C 2 -C 18  alkenyl, C 2 -C 18  alkynyl, C 3 -C 18  cycloalkyl, C 1 -C 18  heterocycloalkyl, C 5 -C 18  aryl, C 5 -C 18  aryloxy, or C 1 -C 18  heteroaryl; and 
 A represents an acidic ionizable modifier moiety or a dual charge ionizable modifier moiety. 
 
       
     
     
         19 . The high purity chromatographic material of  claim 18 , wherein the concentration of ionizable modifier in the high purity chromatographic material is less than 0.5 μmol/m 2  of the specific surface area. 
     
     
         20 . The high purity chromatographic material of  claim 18 , wherein the ratio of the hydrophobic surface group:ionizable modifier is from about 2.5:1 to about 35:1. 
     
     
         21 . The high purity chromatographic material of  claim 18 , further comprising a chromatographic core material. 
     
     
         22 . The high purity chromatographic material of  claim 21 , wherein the chromatographic core material is a silica material or a hybrid inorganic/organic material. 
     
     
         23 . The high purity chromatographic material of  claim 22 , wherein the chromatographic core material is a superficially porous material. 
     
     
         24 . The high purity chromatographic material of  claim 18 , wherein the hydrophobic surface group is a C4 to C30 bonded phase, an aromatic, a phenylalkyl, a fluoro-aromatic, a phenylhexyl, a pentafluorophenylalkyl, or a chiral bonded phase. 
     
     
         25 . The high purity chromatographic material of  claim 24 , wherein the hydrophobic surface group is a C18 bonded phase. 
     
     
         26 . The high purity chromatographic material of  claim 18 , wherein the material is in the form of a particle. 
     
     
         27 . The high purity chromatographic material of  claim 18 , wherein the material has been surface modified. 
     
     
         28 . A method for preparing a high purity chromatographic material according to  claim 21  comprising the steps of:
 a. reacting a chromatographic core with ionizable modifying reagent to obtain a ionizable modified material; and 
 b. reacting the resultant ionizable material with a hydrophobic surface modifying group. 
 
     
     
         29 . A method for preparing a chromatographic material according to  claim 21  comprising the steps of:
 a. reacting a chromatographic core with hydrophobic surface modifying group to obtain a bonded material; and 
 b. reacting the resultant bonded material with an ionizable modifying reagent. 
 
     
     
         30 . A separations device having a stationary phase comprising the high purity chromatographic material of  claim 18 . 
     
     
         31 . A chromatographic column, comprising
 a) a column having a cylindrical interior for accepting a packing material and   b) a packed chromatographic bed comprising the high purity chromatographic material of  claim 18 .   
     
     
         32 . A kit comprising the high purity chromatographic material of  claim 18 , and instructions for use. 
     
     
         33 . A chromatographic device, comprising
 a) an interior channel for accepting a packing material, and   b) a packed chromatographic bed comprising the high purity chromatographic material of  claim 18 .   
     
     
         34 . A method for selectively isolating, separating or purifying a macromolecule of a peptide, protein, nucleic acid, or nucleotide from a sample, the method comprising the steps of:
 a) loading a sample containing the macromolecule onto a chromatographic separations device comprising the high purity chromatographic material of  claim 18 , such that the macromolecule is selectively adsorbed onto the high purity chromatographic material; and   b) eluting the adsorbed macromolecule from the high purity chromatographic material, thereby selectively isolating the macromolecule from the sample.   
     
     
         35 . The method of  claim 34 , wherein the chromatographic separations device is a device is selected from the group consisting of a chromatographic column, a thin layer plate, a filtration membrane, a microfluidic separation device, a sample cleanup device, a solid support, a solid phase extraction device, a microchip separation device, and a microtiter plate. 
     
     
         36 . The method of  claim 34 , wherein the peptide, protein, nucleic acid, or nucleotide is selected from the group consisting of a peptide, a polypeptide, a phosphopeptide, a glycopeptide, a protein, a glycoprotein, an antibody, a phosphoprotein, a nucleic acid, an oligonucletoide, a polynucleotide, and mixtures thereof. 
     
     
         37 . The method of  claim 34 , further comprising the step of preparing the sample by treating a mother sample to a secondary chromatographic means to obtain the sample. 
     
     
         38 . The method of  claim 34 , further comprising the step of treating the macromolecules eluted in step b with a secondary chromatographic means to further isolate, purify, or separate the macromolecules.