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Ion exchange chromatographic separation and isolation of oligosaccharides of intact low-molecular-weight heparin for the determination of their anticoagulant and anti-inflammatory properties

Citation

Shastri, MD and Johns, C and Hutchinson, JP and Khandagale, M and Patel, RP, Ion exchange chromatographic separation and isolation of oligosaccharides of intact low-molecular-weight heparin for the determination of their anticoagulant and anti-inflammatory properties, Analytical and Bioanalytical Chemistry, 405, (18) pp. 6043-6052. ISSN 1618-2650 (2013) [Refereed Article]

Copyright Statement

Copyright 2013 Springer-Verlag Berlin Heidelberg

DOI: doi:10.1007/s00216-013-6996-9

Abstract

It is well known that enoxaparin, a widely used anticoagulant and low-molecular-weight heparin containing a large number of oligosaccharides, possesses anti-inflammatory activity. Whilst enoxaparin has shown promising results in various inflammatory disorders, some of its oligosaccharides have anti-inflammatory properties and others increase the risk of bleeding due to their anticoagulant effects. The aim of this study was to develop an effective ion exchange chromatographic (IC) technique which allows the separation, isolation and, consequently, the identification of different oligosaccharides of enoxaparin with or without anticoagulant activity. The developedmethod utilises a semi-preparative CarboPac PA100 (9250 mm) ion exchange column with sodium chloride gradient elution and UV detection at 232 nm. The method successfully resolved enoxaparin into more than 30 different peaks. IC-derived oligosaccharides with high, moderate, low or no anticoagulant activity were identified using an anti-factor Xa assay. The anti-inflammatory activity of selected oligosaccharides was investigated using the Griess assay. Using this technique, the oligosaccharides of enoxaparin with low or no anticoagulant activity, whilst exhibiting significant antiinflammatory activity, could be fractionated. This technique can provide a platform to identify the oligosaccharides which are devoid of significant anticoagulant activity and are responsible for the therapeutic effects of enoxaparin that have been observed in various inflammatory conditions.

Item Details

Item Type:Refereed Article
Keywords:ion exchange chromatography, enoxaparin, anticoagulant oligosaccharides, non-anticoagulant oligosaccharides, anti-inflammatory activity
Research Division:Medical and Health Sciences
Research Group:Pharmacology and Pharmaceutical Sciences
Research Field:Clinical Pharmacology and Therapeutics
Objective Division:Health
Objective Group:Clinical Health (Organs, Diseases and Abnormal Conditions)
Objective Field:Clinical Health (Organs, Diseases and Abnormal Conditions) not elsewhere classified
Author:Shastri, MD (Mr Madhur Shastri)
Author:Johns, C (Dr Cameron Johns)
Author:Hutchinson, JP (Dr Joseph Hutchinson)
Author:Khandagale, M (Mr Manish Khandagale)
Author:Patel, RP (Dr Rahul Patel)
ID Code:88532
Year Published:2013
Web of Science® Times Cited:11
Deposited By:Pharmacy
Deposited On:2014-02-06
Last Modified:2017-11-02
Downloads:0

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