Ion-pair high-performance liquid chromatography for determining disaccharide composition in heparin and heparan sulphate

Karamanos, Nikos K.; Vanky, Peter; Tzanakakis, George N.; Tsegenidis, T.; Hjerpe, Anders

doi:10.1016/s0021-9673(96)00930-2

Cited by 108 publications

(80 citation statements)

References 40 publications

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“…Basically, the analysis of GAG chain composition begins with the treatment that employs appropriate lyases to produce unsaturated disaccharides; in this case the digestion was carried out using heparinases I, II, III, three enzymes with different substrate specificity able to completely degrade the HE/ HS chains to disaccharide units [14]. Depending on the selected method for the analysis, the digestion may be followed by the labelling with a fluorescent dye, for improvement of measurement sensitivity.…”

Section: Resultsmentioning

confidence: 99%

New electrophoretic and chromatographic techniques for analysis of heparin and heparan sulfate

et al. 2008

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Heparin (HE) and heparan sulfated glycosaminoglycans are well-known mediators of tissue development, maintenance and functions; the activities of these polysaccharides are depending mainly on their sulfate substitutions. The HE structure is also a very important feature in antithrombotic drug development, since the antithrombin binding site is composed by sequences of a specific sulfation pattern. The analysis of disaccharide composition is then a fundamental point of all the studies regarding HE/heparan sulfate glycosaminoglycan (and thereby proteoglycan) functions. The present work describes two analytical methods to quantify the disaccharides constituting HE and heparan sulfate chains. The use of PAGE of fluorophore-labeled saccharides and HPLC coupled with a fluorescence detector allowed in one run the identification of 90-95% of HE disaccharides and 74-100% of rat kidney purified heparan sulfate. Moreover, the protocol here reported avoid the N-sulfation disaccharides degradation, which may affect N-sulfated/N-acetylated disaccharides ratio evaluation. These methods could be also very important in clinical treatments since they are useful for monitoring the availability kinetics of antithrombotic drugs, such as low-molecular-weight HEs.

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Section: Resultsmentioning

confidence: 99%

New electrophoretic and chromatographic techniques for analysis of heparin and heparan sulfate

et al. 2008

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“…In order to obtain better separation, some new techniques have been recently developed in research field of liquid chromatography. These are micellar electrokinetic capillary chromatography (MECC) [77], high-speed counter-current chromatography (HSCCC), low-pressure size-exclusion chromatography (SEC) [78], reversed-phase ion-pairing HPLC (RP-IPC-HPLC) [79,80], and strong anion-exchange HPLC (SAX-HPLC) [81]. They will provide new opportunities for good separation for some specific extracts of some herbal medicines.…”

Section: High-performance Liquid Chromatographymentioning

confidence: 99%

Quality control of herbal medicines

Liang

Xie²,

Chan

2004

Journal of Chromatography B

327

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Different chromatographic and electrophoretic techniques commonly used in the instrumental inspection of herbal medicines (HM) are first comprehensively reviewed. Chemical fingerprints obtained by chromatographic and electrophoretic techniques, especially by hyphenated chromatographies, are strongly recommended for the purpose of quality control of herbal medicines, since they might represent appropriately the "chemical integrities" of the herbal medicines and therefore be used for authentication and identification of the herbal products. Based on the conception of phytoequivalence, the chromatographic fingerprints of herbal medicines could be utilized for addressing the problem of quality control of herbal medicines. Several novel chemometric methods for evaluating the fingerprints of herbal products, such as the method based on information theory, similarity estimation, chemical pattern recognition, spectral correlative chromatogram (SCC), multivariate resolution, etc. are discussed in detail with examples, which showed that the combination of chromatographic fingerprints of herbal medicines and the chemometric evaluation might be a powerful tool for quality control of herbal products.

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“…When these lyases are used in combination, the polymeric chain is cleaved to a large extent (> 90%) to A-disaccharides and the disaccharide composition can therefore be obtained. This approach has recently been used to separate and determine all commercially available heparin-' and HS-derived A-disaccharides by a sensitive ion-pair HPLC method [12]. The first CE method on this topic was described in 1991 [55] where A-disaccharides, obtained by enzymic depolymerization of heparin, were separated in an alkaline borate buffer, pH 8.8, using normal polarity, but the migrations of C-4 and C-6 sulfated species were close to each other.…”

Section: Depolymerized Hs and Heparinmentioning

confidence: 99%

A survey of methodological challenges for glycosaminoglycan/proteoglycan analysis and structural characterization by capillary electrophoresis

Karamanos

Hjerpe

1998

Electrophoresis

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Proteoglycans participate and regulate several physiological processes via their glycosaminoglycan constituents. For a deeper understanding of how they interact with extracellular ligands as well as with cell bound effector molecules, the fine chemical structures of their glycosaminoglycan chains must be elucidated. Lately developed capillary electrophoretic techniques is a powerful analytical tool for the analysis of glycosaminoglycans, combining a high resolving power with sensitive detection. In this review we describe how depolymerized and intact glycosaminoglycans/proteoglycans can be characterized by capillary electrophoresis, relating these analyses to their possible biological significance. Conditions for running these separations and the detection systems for particular applications are also summarized.

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Ion-pair high-performance liquid chromatography for determining disaccharide composition in heparin and heparan sulphate

Cited by 108 publications

References 40 publications

New electrophoretic and chromatographic techniques for analysis of heparin and heparan sulfate

New electrophoretic and chromatographic techniques for analysis of heparin and heparan sulfate

Quality control of herbal medicines

A survey of methodological challenges for glycosaminoglycan/proteoglycan analysis and structural characterization by capillary electrophoresis

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