Characterization of Currently Marketed Heparin Products: Reversed-Phase Ion-Pairing Liquid Chromatography Mass Spectrometry of Heparin Digests
Characterization of Currently Marketed Heparin Products: Reversed-Phase Ion-Pairing Liquid Chromatography Mass Spectrometry of Heparin Digests
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DOI：
10.1021/ac102301j
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发表时间：
2010-12-01
影响因子：
7.4
通讯作者：
Keire, David A.
中科院分区：
文献类型：
--
作者：
Brustkern, Adam M.;Buhse, Lucinda F.;Keire, David A.
在这里，我们报告的结果，从分析酶消化和反相离子对液相色谱质谱法活性药物成分（API）、来自六个不同制造商的未分级肝素（UFH）和一个USP标准样品的RPIP-LC-MS 18柱，己胺为离子对试剂，乙腈梯度洗脱，分离裂解酶I和III消化肝素产生的双糖（EC 4 2 2 7和4 2 2 8）通过电喷雾电离（ESI）接口引入离子阱质谱仪之前提取的离子色谱图（EIC）观察到8种二糖，并且从20个UFH样品的质谱中观察到类似的组成分布。此外，RPIP-LC-MS测定中获得的曲线对肝素中污染物、过硫酸化硫酸软骨素A（OSCS）的存在敏感
Here we report results from the analyses by enzymatic digestion and reversed-phase ion pairing liquid chromatography mass spectrometry (RPIP-LC-MS) of active pharmaceutical ingredient (API) unfractionated heparins (UFHs) from six different manufacturers and one USP standard sample We employed a reverse phase ionpairing chromatography method using a C-18 column and hexylamine as the ion pairing reagent with acetonitrile gradient elution to separate disaccharides generated from the digestion of the heparins by lyase I and III (E C 4 2 2 7 and 4 2 2 8) before introduction into an ion-trap mass spectrometer by an electrospray ionization (ESI) interface Extracted ion chromatograms (EICs) were used to determine the relative abundance of the disaccharides by mass spectrometry Eight disaccharides were observed and a similar composition profile was observed from digests of 20 UFH samples The compositional profile determined from these experiments provides a measure of the norm and range of variation in "good" heparin to which future preparations can be compared Furthermore, the profile obtained in the RPIP-LC-MS assay is sensitive to the presence of the contaminant, oversulfated chondroitin sulfate A (OSCS), in heparin