Chemical microdetermination of heparin in plasma

Toshihiko Toida; Yong Huang; Yukio Washio; Tomohiro Maruyama; Hidenao Toyoda; Toshio Imanari; Robert J. Linhardt

doi:10.1006/abio.1997.2277

Chemical microdetermination of heparin in plasma

Toshihiko Toida, Yong Huang, Yukio Washio, Tomohiro Maruyama, Hidenao Toyoda, Toshio Imanari, Robert J. Linhardt

Research output: Contribution to journal › Article › peer-review

21 Scopus citations

Abstract

A new, simple, and highly sensitive method for the determination of heparin has been established. Heparin was first converted into unsaturated disaccharides through the action of heparin lyases I, II, and III. A major trisulfated unsaturated disaccharide product results, consistent with structural analysis of a number of pharmaceutical heparins using one- and two-dimensional ¹H NMR spectroscopy. This disaccharide was analyzed by HPLC using fluorometric postcolumn derivatization. The correlation between the amount of this trisulfated unsaturated disaccharide and anticoagulant activity of heparin as measured by anti-IIa was determined. The analysis of these pharmaceutical heparins showed a linear correlation between both HPLC and bioassay. This HPLC method was then applied to a pharmacokinetic study of heparin intravenously administered to rabbits.

Original language	English
Pages (from-to)	219-226
Number of pages	8
Journal	Analytical Biochemistry
Volume	251
Issue number	2
DOIs	https://doi.org/10.1006/abio.1997.2277
State	Published - 5 Sep 1997
Externally published	Yes

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AU - Toida, Toshihiko

AU - Huang, Yong

AU - Washio, Yukio

AU - Maruyama, Tomohiro

AU - Toyoda, Hidenao

AU - Imanari, Toshio

AU - Linhardt, Robert J.

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AB - A new, simple, and highly sensitive method for the determination of heparin has been established. Heparin was first converted into unsaturated disaccharides through the action of heparin lyases I, II, and III. A major trisulfated unsaturated disaccharide product results, consistent with structural analysis of a number of pharmaceutical heparins using one- and two-dimensional 1H NMR spectroscopy. This disaccharide was analyzed by HPLC using fluorometric postcolumn derivatization. The correlation between the amount of this trisulfated unsaturated disaccharide and anticoagulant activity of heparin as measured by anti-IIa was determined. The analysis of these pharmaceutical heparins showed a linear correlation between both HPLC and bioassay. This HPLC method was then applied to a pharmacokinetic study of heparin intravenously administered to rabbits.

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Chemical microdetermination of heparin in plasma

Abstract

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