High-performance liquid affinity chromatography of nucleosides, nucleotides and carbohydrates with boronic acid-substituted microparticulate silica

Glad, Magnus; Ohlson, Sten; Hansson, Lennart; Månsson, Mats-Olle; Mosbach, Klaus

doi:10.1016/s0021-9673(00)84944-4

Cited by 115 publications

(35 citation statements)

References 22 publications

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“…A specially designed boronate affinity solid phase, immobilized 3-amino-phenyl boronic acid, has been developed by Glad et al to achieve complete separation of adenosine phosphates at pH 7.5 [23]. On our synthetic columns, the three phosphates of adenosine are well resolved at a significantly lower pH.…”

Section: Hplc Evaluation Of Boronate Affinitymentioning

confidence: 99%

Synthesis of sulfonamide- and sulfonyl-phenylboronic acid-modified silica phases for boronate affinity chromatography at physiological pH

Pennington

Stobaugh

et al. 2008

Analytical Biochemistry

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Two new types of boronate affinity solid phases were synthesized and characterized. The materials were prepared by silylation of porous silica gel with monochlorosilane derivatives containing synthetic sulfonyl-and sulfonamide-substituted phenylboronic acids. The new solid phases were evaluated for boronate affinity chromatography with aryl and alkyl cis-diol compounds, and found suitable for the retention of cis-diols under acidic conditions. Significant correlations between the retention factor (K) and the pH of the mobile phase demonstrate that the binding of cis-diols to the solid phases is best rationalized by chelation. Based on the low ionization constant, caused by the electron-withdrawing effects of the sulfonyl and sulfonamide groups, these media display an enhanced affinity for cis-diols as compared to unsubstituted phenylboronic acid. Utilizing Isocratic elution, a mixture of various biologically relevant L-tyrosines, L-DOPA and several catecholamines were resolved with a mobile phase composed of 0.05 M, pH5.5 phosphate buffer. Mono-, di-and triphosphates of adenosine were also separated at pH 6.0. Hence, the new boronate solid phase offers efficient affinity separation and purification of cis-diol containg molecules under rather mild pH regions.

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Section: Hplc Evaluation Of Boronate Affinitymentioning

confidence: 99%

Synthesis of sulfonamide- and sulfonyl-phenylboronic acid-modified silica phases for boronate affinity chromatography at physiological pH

Pennington

Stobaugh

et al. 2008

Analytical Biochemistry

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“…Since the occurrence of nucleosides in physiological fluids turned out to be predicative for the diagnosis of pathobiochemical processes, the on-line System is the method of choice, äs it is particularly suitable for automatization and applicable for routine clinical analysis. As affinity chromatography on boronic acid-functionalized polymers is also well known for the pre-purification of catecholamines in physiological fluids (34)(35)(36)(37)(38), for the Separation of ribonucleosides and ribonucleotides from the corresponding deoxy-compounds (22,39,40), the Separation of RNA from DNA (41,42) and the Separation of queuine-containing tRNA from non-queuine tRNA (41) or aminoacyl-tRNA from uncharged tRNA (41,42) the use of boronate HPLAC in combination with the presented column switching technique offers a wide ränge of application.…”

Section: Discussionmentioning

confidence: 99%

“…Encouraged by the publication of a boronic acidfunctionalized silica (22), which is particular suitable for HPLC, we attempted to transfer the commonly used urinary clean-up procedure from Iow pressure column chromatography to HPLC conditions thus applying a method known s high-performance liquid (ligand) affinity chromatography (HPLAC) (23).…”

Section: Introductionmentioning

confidence: 99%

Development of a Chromatographic Method for the Quantitative Determination of Minor Ribonucleosides in Physiological Fluids. Characterization and quantitative determination of minor ribonucleosides in physiological fluids, Part I

Hagemeier¹,

Kemper²,

Ks³

et al. 1984

Clinical Chemistry and Laboratory Medicine

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The authors wish to congratulate Professor Walther Lamprecht, Hannover, on the occasion of his sixtieth birthday Summary:We describe an on-line multi-cplumn high performance liquid Chromatographie method for the selective clean-up and analysis of major and minor ribonucleosides in physiological fluids. Quantitative data obtained for the determination of some methylated ribonucleosides in human urines are compared with those obtained with the traditional off-line method.The on-line technique developed in our laboratory is distinguished from the off-line method by the following features:(1) Sample clean-up and analysis of the target-compounds can easily be automatized, (2) Total time of analysis, for example of urinary ribonucleosides, is decreased to 35 minutes, (3) Laborious and error-prone evaporation and redissolution Steps are avoided, (4) Reliability of the overall analytical System can be controlled with ease, (5) Small sample-volumes can be applied directly, (6) Sensitive samples can be processed very rapidly under mild conditions, (7) Results obtained with the -line and off-line-techniques compare well. Hagemeier, Kemper, Boos and Schlimme: Quantitative determination of minor ribonucleosides (5) Kleine Probenvolumina können direkt appliziert werden, (6) Empfindliche Proben können in kürzester Zeit unter schonenden Bedingungen analysiert werden, (7) Die mit der "on-line" wie mit der "off-line"-Technik erhaltenen Ergebnisse zeigen eine gute ÜbereinŜ timmung. Entwicklung einer chromatographischen Methode zur quantitativen Bestimmung seltener Ribonucleoside in physiologischen Flüssigkeiten Charakterisierung und quantitative Bestimmung von seltenen Ribonucleosiden in physiologischen Flüssig

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“…The connection with NAD is through the exocyclic N of adenine. The progress of the synthesis and the purity of the bis-NAD analogues could be followed by HPLAC on a column of silica-bound boronic acid (14). Immobilization.…”

Section: Methodsmentioning

confidence: 99%

Site to Site Directed Immobilization of Enzymes With Bis-Nad Analogues

Månsson¹,

Siegbahn²,

Mosbach³

1983

Affinity Chromatography and Biological Recognition

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Lactate dehydrogenase (L-lactate:NAD' oxidoreductase, EC 1.1.1.27) and alcohol dehydrogenase (alcohol: NAD' oxidoreductase, EC 1.1.1.1) have been crosslinked with glutaraldehyde on agarose beads. The crosslinking was performed while the two enzymes were spatially arranged with their active sites facing one another with the aid of a bis-NAD analogue. Subsequently the bis-NAD analogue was allowed to diffuse out. By using a third enzyme, lipoamide dehydrogenase (NADH:lipoamide oxidoreductase, EC 1.6.4.3), which was also coupled to the same beads and which competes with lactate dehydrogenase for the NADH produced by alcohol dehydrogenase, the effect of site-tosite directed immobilization was studied. It was found that much more NADH than was theoretically expected (50% instead of 19% of produced NADH) was oxidized by lactate dehydrogenase, which indicates that the NADH was preferentially channeled to lactate dehydrogenase due to the juxtapositioned active sites of the two enzymes.Several coimmobilized multistep enzyme systems have been described in the literature (1-6). For example, an immobilized system composed of the sequence malate dehydrogenase/citrate synthase showed a consistantly higher overall steady-state rate (6). More recently, a four-enzyme sequence (7) and even the enzymes of a complete metabolic cycle, the urea cycle, have been coimmobilized to supports (8). In the latter case, the immobilized cyclic enzyme system again was more efficient than the corresponding soluble system. These effects have been partly attributed to the close proximity of the enzymes and partly to the diffusional restrictions imposed by the Nernst unstirred layer around the enzymes (2, 9).

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High-performance liquid affinity chromatography of nucleosides, nucleotides and carbohydrates with boronic acid-substituted microparticulate silica

Cited by 115 publications

References 22 publications

Synthesis of sulfonamide- and sulfonyl-phenylboronic acid-modified silica phases for boronate affinity chromatography at physiological pH

Synthesis of sulfonamide- and sulfonyl-phenylboronic acid-modified silica phases for boronate affinity chromatography at physiological pH

Development of a Chromatographic Method for the Quantitative Determination of Minor Ribonucleosides in Physiological Fluids. Characterization and quantitative determination of minor ribonucleosides in physiological fluids, Part I

Site to Site Directed Immobilization of Enzymes With Bis-Nad Analogues

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