Rapid and sensitive determination of enzymatic degradation products of isomeric chondroitin sulfates by high-performance liquid chromatography

Lee, George Jia-Long; Evans, James E.; Tieckelmann, Howard

doi:10.1016/s0378-4347(00)81202-5

Cited by 37 publications

(3 citation statements)

References 23 publications

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“…However, the majority of these methods have proved unsatisfactory. Chromatography on silica gel columns is performed with a ternary elution mixture of dichloromethäne, methanol and aqueous buffer Solutions which is difficult to use (12). The disadvantages of ion-pair chromatography using reversed-phase columns are löng retention times and pronounced sensitivity to minor variations in the eluent (13).…”

Section: High Performance Liquid Chromatographymentioning

confidence: 99%

High Performance Liquid Chromatographie Assay of Disaccharides and Oligosaccharides Produced by the Digestion of Glycosaminoglycans with Chondroitin Sulphate Lyases

Gurr¹,

G²,

Tunn³

et al. 1985

Clinical Chemistry and Laboratory Medicine

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Gurr et al.: HPLC assay of disaccharides and oligosaccharides 77 Summary: In high perfonnance liquid Chromatographie procedures hitherto described, SiO 2 , NH 2 and RP columns have been used for the analysis of disaccharides produced by % the digestion of glycosaminoglycans with the chondroitin sulphate lyases AC and ABC. The use of a potent anion exchanger offers the following advantages over these columns: superior Separation chäracteristics for non-sulphated disaccharides, and improved column perfonnance, coupled with more stable analytical conditions. Elution with dilute saline Solutions permits Separation of the two non-sulphated disaccharides from chondroitin and hyaluronate.The sequential application of chondroitinase AC and ABC permits the determination of hyaluronate, the chondroitin sulphate isomers and the dermatan sulphate isomers by high perfonnance liquid Chromatographie Separation of the products of enzymatic hydrolysis. In a previously described method, hyaluronate lyase was used for the determination of hyaluronate. It has been found, however, that omission of the hyaluronate lyase step results in superior accuracy in the high performance liquid Chromatographie Separation of the nonsulphated disaccharides.The enzymatic analysis of human articular cartilage glycosaminoglycans has repeatedly yielded a fraction which is not digestable by chondroitinase AC, but is completely digestable by chondroitinase ABC. More extensive characterization has disclosed that this fraction differs structurally from chondroitin sulphate. Enzymatic characterization indicates that it should presumably be assigned to dermatan sulphate. Hochleistungsflüssigkeitschromatographische Bestimmung von Di-und Oligosacchariden aus dem Abbau von Glykosaminoglykanen mit ChondroitinsulfatlyasenZusaHunenfassung: Bei den bisher beschriebenen hochleistungsflüssigkeitschromatographischen Verfahren zur Analyse von Disacchariden aus dem Abbau von Glykosaminoglykanen mit den Chondroitinsulfatlyasen AC und ABC wurden SiO 2 -, NHk-und RP-Säulen verwendet. Gegenüber diesen Säulen bietet die Verwendung eines starken Anionenaustauschers folgende Vorteile: bessere Trenneigenschaften im Bereich der unsulfatierten Disaccharide und höhere Laufleistung pro Säulenfällung bei stabileren Analysenbedingungen. Die Elution mit verdünnten Kochsalzlösungen ermöglicht die Trennung der beiden unsulfatierten Disaccharide aus Chondroitin und Hyaluronat.Durch die sequentielle Anwendung von Chondroitinase AC und ABC können durch hochleistungsflüssigkeits-chromatographische Trennung der Metabolite Hyaluronat, die Chondroitinsulfatisomere sowie die Dermatansulfatisomere bestimmt werden. Verglichen mit einer früher beschriebenen Methode, bei der zur Hyaluronatbestimmung Hyaluronatlyase verwendet wurde, zeigt sich, daß ohne den Hyaluronatlyase-Schritt durch hochleistungsflüssigkeitschromatographische Trennung der unsulfatierten Disaccharide eine bessere Richtigkeit erreicht wird.

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Section: High Performance Liquid Chromatographymentioning

confidence: 99%

High Performance Liquid Chromatographie Assay of Disaccharides and Oligosaccharides Produced by the Digestion of Glycosaminoglycans with Chondroitin Sulphate Lyases

Gurr¹,

G²,

Tunn³

et al. 1985

Clinical Chemistry and Laboratory Medicine

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“…(i) Chondroitinase ABC-HPLC: i mg of sample was digested with 12 unit of chondroitinase ABC (Seikagaku Kogyo Ltd, Tokyo, Japan). Analysis ofthe digest was carried out by using the method of Lee, Evans & Pieckelmann (1978).…”

Section: Determination Of Gags By Enzymatic Digestion-high Performancmentioning

confidence: 99%

Chemical components of jelly-like matrix in digital mucous cyst

et al. 1985

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Summary Jelly‐like matrix taken from a lesion of digital mucous cyst (DMC) was analysed with cellulose acetate paper electrophoresis and an enzymatic digestion‐high performance liquid chromato‐graph method and compared qualitatively and quantitatively with that from a ganglion. The ratio of glycoconjugate to the total solid components was extremely high in matrix from DMC, while only less than 1% in that from ganglion. In the former the greater part of glycoconjugate was glycosaminoglycans (GAGs) consisting mainly of hyaluronic acid, while in the latter glycoprotein dominated and the amount of GAGs was very small.

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“…One alternative method is the separation of isomers via LC/ HPLC 24,25 or electrophoresis. 26,27 However, the identication and quantication by these approaches take several minutes per sample, usually involve derivatization of the analytes, and require normally pure reference compounds for calibration and identication of the compounds in the sample.…”

Section: Introductionmentioning

confidence: 99%

Comparison of ion mobility-mass spectrometry and pulsed-field gradient nuclear magnetic resonance spectroscopy for the differentiation of chondroitin sulfate isomers

et al. 2016

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Rapid and sensitive determination of enzymatic degradation products of isomeric chondroitin sulfates by high-performance liquid chromatography

Cited by 37 publications

References 23 publications

High Performance Liquid Chromatographie Assay of Disaccharides and Oligosaccharides Produced by the Digestion of Glycosaminoglycans with Chondroitin Sulphate Lyases

High Performance Liquid Chromatographie Assay of Disaccharides and Oligosaccharides Produced by the Digestion of Glycosaminoglycans with Chondroitin Sulphate Lyases

Chemical components of jelly-like matrix in digital mucous cyst

Comparison of ion mobility-mass spectrometry and pulsed-field gradient nuclear magnetic resonance spectroscopy for the differentiation of chondroitin sulfate isomers

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