Publication: Development and validation of an ion-exchange chromatography method for heparin and its impurities in heparin products
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Issued Date
2014-01-01
Resource Type
ISSN
16159314
16159306
16159306
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2-s2.0-84910020358
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Separation Science. Vol.37, No.22 (2014), 3195-3204
Suggested Citation
Sumate Thiangthum, Yvan Vander Heyden, Wolfgang Buchberger, Johan Viaene, Brompoj Prutthiwanasan, Leena Suntornsuk Development and validation of an ion-exchange chromatography method for heparin and its impurities in heparin products. Journal of Separation Science. Vol.37, No.22 (2014), 3195-3204. doi:10.1002/jssc.201400348 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/33598
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Title
Development and validation of an ion-exchange chromatography method for heparin and its impurities in heparin products
Abstract
© 2014 WILEY-VCH Verlag GmbH & Co. KGaA. An anion-exchange liquid chromatography method for the determination of heparin and its impurities (dermatan sulfate and oversulfated chondroitin sulfate) was developed using chemometric-assisted optimization, including multivariate experimental design and response surface methodology. The separation of heparin, dermatan sulfate, and oversulfated chondroitin sulfate (Rs above 2.0) was achieved on a Dionex RF IC IonPac AS22 column with a gradient elution of 10-70% of 2.5 M sodium chloride and 20 mM Tris phosphate buffer (pH 2.1) at a flow rate of 0.6 mL/min and UV detection at 215 nm. Method validation shows good linearity (r > 0.99), acceptable precision (%relative standard deviations <11.4%) and trueness (%recovery of 92.3-103.9%) for all analytes. The limits of detection for dermatan sulfate and oversulfated chondroitin sulfate are equivalent to 0.11% w/w (10.5 μg/mL) and 0.07% w/w (7.2 μg/mL), while the limits of quantification are 0.32% w/w (31.5 μg/mL) and 0.22% w/w (22.0 μg/mL) relative to heparin, respectively. The method is specific for heparin, dermatan sulfate, and oversulfated chondroitin sulfate without interference from mobile phase and sample matrices and could be used for accurate quantitation the drug and its impurities in a single run. Applications of the method reveal contents of heparin between 90.3 and 97.8%. Dermatan sulfate and oversulfated chondroitin sulfate were not detected in any of the real-life samples.