Process for Analyzing, for Separating, and for Isolating Individual Polar Protic Monomers and/or Oligomers

Abstract

An improved process for separating and isolating individual polar protic monomer(s) and/or oligomer(s) on the basis of degree of polymerization. A liquid sample containing polar protic monomer(s) and/or oligomer(s) is introduced into a liquid chromatography (LC) column packed with a polar bonded stationary chromatographic phase. The individual polar protic monomer(s) and/or oligomer(s) are separated via a binary mobile phase elution. One or more individual fractions containing the monomer(s) and/or oligomer(s) are eluted. The polar protic monomer(s) and/or oligomer(s) may be proanthocyanidins, hydrolyzable tannins, oligosaccharides, oligonucleotides, peptides, acrylamides, polysorbates, polyketides, poloxarners, polyethylene glycols, polyoxyethylene alcohols or polyvinyl alcohols. The binary mobile phase comprises an A phase consisting essentially of a polar aprotic solvent and a B phase consisting essentially of a polar protic solvent. A process for separating and isolating xanthine(s) (e.g., caffeine and theobromine) from polar protic monomer(s) and/or oligomer(s). A liquid sample containing xanthine(s) and polar protic monomer(s) and/or oligomer(s) is introduced into an LC column packed with a polar bonded stationary chromatographic phase. The xanthines are separated via an isocratic mobile phase elution, and one or more individual fractions containing the xanthines are eluted.

1 - 40 . (canceled) 
   
   
       41 . A normal phase high performance liquid chromatographic process for separating, eluting, and isolating individual polar protic monomer(s) and/or polar protic oligomer(s) comprises the steps of:
 (a) introducing a liquid sample containing the polar protic monomer(s) and/or the polar protic oligomer(s) into a liquid chromatography column packed with a polar bonded stationary phase selected from the group consisting of a diol phase, a glycerol phase, an amino phase, a cyano phase, a trimethylsilyl phase, a dimethylsilyl phase, a halogenated phase, and a nitro phase;   (b) passing a binary mobile phase comprising an A phase consisting essentially of a polar aprotic solvent and a B phase consisting essentially of a polar protic solvent through the packed column to elute the individual polar protic monomer(s) and/or the individual polar protic oligomer(s) on the basis of degree of polymerization;   (c) isolating one or more individual fractions containing the polar protic monomer(s) and/or the polar protic oligomer(s); and   (d) optionally recovering the isolated polar protic monomer(s) and/or the isolated polar protic oligomer(s).   
   
   
       42 . The process of  claim 41 , wherein the polar protic oligomer(s) are proanthocyanidins, hydrolyzable tannins, oligosaccharides, oligonucleotides, peptides, acrylamides, polysorbates, polyketides, poloxamers, polyethylene glycols, polyethylene alcohols, or polyvinyl alcohols. 
   
   
       52 . The process of claim  51 , wherein the proportion of polar aprotic solvent in the A phase is greater than 50% by volume; wherein the proportion of the polar protic solvent in the B phase is greater than 50% by volume; and wherein the remainder of the A phase and the remainder of the B phase is any mineral acid, organic acid, and/or water. 
   
   
       53 . The process of  claim 52 , wherein the proportion of the polar aprotic solvent in the A phase is greater than 90% by volume; wherein the proportion of the polar protic solvent in the B phase is greater than 90% by volume; and wherein the organic acid is acetic acid. 
   
   
       54 . The process of  claim 52 , wherein the A phase consists essentially of a mixture of about 98% acetonitrile and about 2% acetic acid (v/v). 
   
   
       55 . The process of  claim 52 , wherein the A phase consists essentially of a mixture of about 99% acetonitrile and 1% acetic acid (v/v). 
   
   
       56 . The process of  claim 52 , wherein the B phase consists essentially of a mixture of about 95% methanol, about 3% water, and about 2% acetic acid (v/v/v). 
   
   
       57 . The process of  claim 52 , wherein the B phase consists essentially of a mixture of about 95% methanol, about 4% water, and about 1% acetic acid (v/v/v). 
   
   
       58 . The process of  claim 41 , wherein the stationary phase has a particle size from about 3 μm to about 10 μm.