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Molecules 2017, 22(7), 1214; doi:10.3390/molecules22071214

Molecular Weights of Bovine and Porcine Heparin Samples: Comparison of Chromatographic Methods and Results of a Collaborative Survey

1
Istituto di Ricerche Chimiche e Biochimiche “G. Ronzoni” via G. Colombo 81, 20133 Milano, Italy
2
Department of Chemistry, University of Pavia, viale Taramelli 12, 27100 Pavia, Italy
3
U.S. Pharmacopeial Convention (USP), 12601 Twinbrook Parkway, Rockville, MD 20852, USA
4
Center for Scientific Review (CSR), National Institutes of Health, 6701 Rockledge Dr. Rm. 4187, Bethesda, MD 20892, USA
5
Institute of Pharmaceutical Science, Franklin Wilkins Building, King’s College London, 150 Stamford St., London SE1 9NH, UK
*
Author to whom correspondence should be addressed.
Received: 24 May 2017 / Accepted: 15 July 2017 / Published: 19 July 2017
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Abstract

In a collaborative study involving six laboratories in the USA, Europe, and India the molecular weight distributions of a panel of heparin sodium samples were determined, in order to compare heparin sodium of bovine intestinal origin with that of bovine lung and porcine intestinal origin. Porcine samples met the current criteria as laid out in the USP Heparin Sodium monograph. Bovine lung heparin samples had consistently lower average molecular weights. Bovine intestinal heparin was variable in molecular weight; some samples fell below the USP limits, some fell within these limits and others fell above the upper limits. These data will inform the establishment of pharmacopeial acceptance criteria for heparin sodium derived from bovine intestinal mucosa. The method for MW determination as described in the USP monograph uses a single, broad standard calibrant to characterize the chromatographic profile of heparin sodium on high-resolution silica-based GPC columns. These columns may be short-lived in some laboratories. Using the panel of samples described above, methods based on the use of robust polymer-based columns have been developed. In addition to the use of the USP’s broad standard calibrant for heparin sodium with these columns, a set of conditions have been devised that allow light-scattering detected molecular weight characterization of heparin sodium, giving results that agree well with the monograph method. These findings may facilitate the validation of variant chromatographic methods with some practical advantages over the USP monograph method. View Full-Text
Keywords: bovine heparin; porcine heparin; molecular weight; size exclusion chromatography; pharmacopeia bovine heparin; porcine heparin; molecular weight; size exclusion chromatography; pharmacopeia
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MDPI and ACS Style

Bertini, S.; Risi, G.; Guerrini, M.; Carrick, K.; Szajek, A.Y.; Mulloy, B. Molecular Weights of Bovine and Porcine Heparin Samples: Comparison of Chromatographic Methods and Results of a Collaborative Survey. Molecules 2017, 22, 1214.

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