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Anal Bioanal Chem (2014) 406:48154823 DOI 10.1007/s00216-014-7940-3
FEATURE ARTICLE
USP compendial methods for analysis of heparin: chromatographic determination of molecular weight distributions for heparin sodium
Barbara Mulloy & Alan Heath & Zachary Shriver &
Fabian Jameison & Ali Al Hakim & Tina S. Morris &
Anita Y. Szajek
Published online: 24 June 2014# Springer-Verlag Berlin Heidelberg 2014
AbbreviationsBST Broad standard tableFDA Food and Drug Administration HP Heparin sodium drug productMn Number-average molecular weight
Mp Peak molecular weightMw Weight-average molecular weight
MW Molecular weightRSD Relative standard deviation
SD Standard deviationSEC Size-exclusion chromatography USP United States Pharmacopeia
Introduction
Heparin is a polysaccharide product isolated from glycosaminoglycans of porcine mucosa (or occasionally other tissues and species). It is a linear non-uniform polymer consisting of alternating glucosamine and uronic acid monosaccharide residues and is highly sulfated [1]. Heparin sodium drug product (HP) used in medicine consists of chains with molecular weight (MW) ranging from under 5,000 to over 50,000 [2].
Although HP has been used as an injectable antithrombotic medicine for more than 70 years [3], many aspects of its structure and purity, including its MW, have not been specified by public standards until recent years. In 2008, a number of HP lots associated with severe adverse effects, including fatalities, were found to have been contaminated with oversulfated chondroitin sulfate [4]. This incident led to thorough revision of compendial standards worldwide. In the USA, the Food and Drug Administration (FDA) encouraged the inclusion of enhanced standards for purity and identity in the relevant monographs of the United States Pharmacopeia (USP) including acceptance criteria for MW distribution.
Heparin originates in mast cell granules, in which it is the polysaccharide part of the proteoglycan serglycin [5]. On degranulation, heparin is released from mast cells and is broken down by endogenous heparanase to fragments, most of which are between 5 and 30 kDa in mass [6]. It is reasonable to expect that variations in manufacturing procedures will result in corresponding differences in the MW distribution of the finished HP product. To ensure an acceptable degree of consistency between HP products, and to decide what the
The findings and conclusions presented have not been formally disseminated by the FDA and should not be construed to represent any FDA determination or policy.
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