Procedures for the analysis of cyanogen bromide-activated Sepharose or Sephadex by quantitative determination of cyanate esters and imidocarbonates

Joachim Kohn, Meir Wilchek

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Analytical methods have been developed for the determination of cyanate esters and imidocarbonates, the active species present on CNBr-activated polysaccharide resins. Imidocarbonates are determined by selective acid hydrolysis followed by determination of the liberated ammonia by a modification of the ninhydrin reaction. Cyanate esters are determined by a spectrophotometric procedure based on the reaction with pyridine and employing N,N'-dimethylbarbituric acid as a color-forming reagent. For the determination of the coupling capacity, a procedure is suggested which allows the amount of coupled ligand to be determined directly on the resin and without prior hydrolysis. Using those procedures it was found that the coupling capacity of activated resins toward small ligands can be predicted by determining the amounts of cyanate esters and imidocarbonates present on the resin, and that cyanate esters are predominantly responsible for coupling of ligand to activated Sepharose.

Original languageEnglish (US)
Pages (from-to)375-382
Number of pages8
JournalAnalytical Biochemistry
Volume115
Issue number2
DOIs
StatePublished - Jan 1 1981
Externally publishedYes

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Cyanates
Cyanogen Bromide
Sepharose
Esters
Resins
Ligands
Hydrolysis
Ninhydrin
Acids
Ammonia
Polysaccharides
Color
sephadex

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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abstract = "Analytical methods have been developed for the determination of cyanate esters and imidocarbonates, the active species present on CNBr-activated polysaccharide resins. Imidocarbonates are determined by selective acid hydrolysis followed by determination of the liberated ammonia by a modification of the ninhydrin reaction. Cyanate esters are determined by a spectrophotometric procedure based on the reaction with pyridine and employing N,N'-dimethylbarbituric acid as a color-forming reagent. For the determination of the coupling capacity, a procedure is suggested which allows the amount of coupled ligand to be determined directly on the resin and without prior hydrolysis. Using those procedures it was found that the coupling capacity of activated resins toward small ligands can be predicted by determining the amounts of cyanate esters and imidocarbonates present on the resin, and that cyanate esters are predominantly responsible for coupling of ligand to activated Sepharose.",
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AU - Wilchek, Meir

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N2 - Analytical methods have been developed for the determination of cyanate esters and imidocarbonates, the active species present on CNBr-activated polysaccharide resins. Imidocarbonates are determined by selective acid hydrolysis followed by determination of the liberated ammonia by a modification of the ninhydrin reaction. Cyanate esters are determined by a spectrophotometric procedure based on the reaction with pyridine and employing N,N'-dimethylbarbituric acid as a color-forming reagent. For the determination of the coupling capacity, a procedure is suggested which allows the amount of coupled ligand to be determined directly on the resin and without prior hydrolysis. Using those procedures it was found that the coupling capacity of activated resins toward small ligands can be predicted by determining the amounts of cyanate esters and imidocarbonates present on the resin, and that cyanate esters are predominantly responsible for coupling of ligand to activated Sepharose.

AB - Analytical methods have been developed for the determination of cyanate esters and imidocarbonates, the active species present on CNBr-activated polysaccharide resins. Imidocarbonates are determined by selective acid hydrolysis followed by determination of the liberated ammonia by a modification of the ninhydrin reaction. Cyanate esters are determined by a spectrophotometric procedure based on the reaction with pyridine and employing N,N'-dimethylbarbituric acid as a color-forming reagent. For the determination of the coupling capacity, a procedure is suggested which allows the amount of coupled ligand to be determined directly on the resin and without prior hydrolysis. Using those procedures it was found that the coupling capacity of activated resins toward small ligands can be predicted by determining the amounts of cyanate esters and imidocarbonates present on the resin, and that cyanate esters are predominantly responsible for coupling of ligand to activated Sepharose.

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