Stationary phase-based two-dimensional chromatography combining both covalent and noncovalent interactions on a single HPLC column

Abstract

A new type of 2-D separation material was synthesized and studied. The material is suitable for 2-D chromatography utilizing both covalent and noncovalent interactions. The first dimension is boronate affinity chromatography, and the second dimension is RP chromatography (or vice versa). The polymeric media were prepared using p-vinylphenylboronic acid as the functional monomer. This monomer was selected due to the presence of the boronic acid group for the cis-diol/boronate interaction in boronate chromatography. Two crosslinkers were evaluated, namely ethylene glycol dimethacrylate and divinylbenzene. The crosslinker content was varied to maximize the polymer strength and the RP performance of the packed column. Several parameters were evaluated to define the optimum for polymer strength and column performance including crosslinker, porogen, initiator, and column-packing parameters. The polymer-based HPLC columns were successful in separating phenol, catechol, dimethylphthalate, and hydroquinone under RP conditions, and thus can be used as an RP HPLC column. The columns were also successful in separating catechol and adenosine under boronate chromatography conditions, and thus can be used as a boronate affinity column. Moreover, the two types of chromatography can be performed consecutively on the same column during one complete chromatographic run, making it a 2-D chromatography. Under these 2-D conditions, the catechol was separated from a mixture of phenol, catechol, dimethylphthalate, and hydroquinone; the adenosine ribonucleoside was separated from a mixture of adenosine ribonucleoside, adenosine deoxyribonucleoside, and uridine deoxyribonucleoside. This type of single-column 2-D HPLC eliminates the requirement of a complex and expensive multidimensional HPLC instrument and provides increased peak capacity for separation.

A Reusable Interface Constructed by 3‐Aminophenylboronic Acid‐Functionalized Multiwalled Carbon Nanotubes for Cell Capture, Release, and Cytosensing

“…The first reports of boric12, 13 and boronic acid13 forming compounds with saccharides appeared in the 1950s, and this pioneering work has since attracted considerable attention. To date, boronic acid chemistry and the pH‐dependent reversible formation of these complexes have been exploited in a number of applications including affinity chromatography to selectively capture and separate cis‐diol‐containing biomolecules,14, 15 such as glycoproteins,16–22 glycosylated peptides,23 enzymes,24 carbohydrates,25, 26 catechols,27, 28 and nucleosides;29–31 capillary electrophoresis,32 aqueous sugar sensors,33 and glycoprotein‐immobilization in cellulose beads 34. Furthermore, because of the wide variety of poly‐ and oligosaccharides localized on the walls of microbial and animal cells which can be targeted by sugar‐specific ligands, we can use the sugar‐specific antibodies and lectins for controllable attachment and detachment of cells 35.…”

A unique boronic acid functionalized monolithic capillary for specific capture, separation and immobilization of cis-diol biomolecules

“…Similar 2D separation capability has been reported, but the separation mechanism is boronate affinity plus reversed-phase separation. 10 As mentioned above, the monolithic capillary is highly hydrophilic, thus the involvement of reversed-phase separation is impossible. Ion exchange mechanism is also impossible, because the boronic acid is uncharged at the elution condition used (pH 2.7).…”

Enrichment of adenosine using thermally responsive chromatographic materials under friendly pH conditions

“…Subsequently, α‐glycosidase was successfully captured by 3‐nitro‐4‐(6‐aminohexylamido)phenylboronic acid grafted on the surface of sepharose‐CL6B by Myohanen and coworkers . Since then, this technique has been exploited for the separation of various cis ‐diol‐containing compounds . The principle relies on reversible covalent complex formation/dissociation between boronic acids and cis ‐diols in alkaline/acidic aqueous solutions .…”