Characterization of currently marketed heparin products: analysis of molecular weight and heparinase-I digest patterns
Characterization of currently marketed heparin products: analysis of molecular weight and heparinase-I digest patterns
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DOI：
10.1007/s00216-011-5362-z
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发表时间：
2011-11-01
影响因子：
4.3
通讯作者：
Keire, David A.
中科院分区：
文献类型：
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作者：
Sommers, Cynthia D.;Ye, Hongping;Keire, David A.
评价了聚丙烯酰胺凝胶电泳法(PAGE)和多角度激光光散射色谱(SEC-MALLS)测定肝素重均相对分子质量(M(W))和多分散度(Pd)的方法。对从供应美国市场的九家制造商获得的一套普通肝素钠(UFH)和低分子肝素(LMWH)样本进行了评估。对于SEC-MALL，我们在评估分子量之前测量了每个样品的水分含量、折射率增量(dN/dc)和第二维里系数(A(2))。对于UFH，M(W)的平均+/-标准差值为16,773+/-797，范围为15,620至18,363(n=20，一式三份)。对于SEC-MALL的低分子肝素，我们测得达肝素、替氮肝素和依诺肝素的M(W)平均值分别为6,717+/-71(n=4)、6,670+/-417(n=3)和3,959+/-145(n=3)。相同的UFH、达肝素、替扎肝素和依诺肝素的PAGE分析结果分别为16,135+/-643(n=20)、5,845+/-45(n=4)、6,049+/-95(n=3)和4,772+/-69(n=3)。这些正交测量是在2008年肝素危机改变了这一药物类别的审查水平后，对正在上市的产品进行大肝素样本集所获得的第一个M(W)结果。在这项研究中，我们将我们的新数据集与10多年前分析的样本进行了比较。此外，我们发现肝素酶消化的UFH和整齐的LMWH样品的PAGE分析产生了特征图谱，为药品质量和一致性的鉴定和评估提供了一种简便的方法。
We evaluated polyacrylamide gel electrophoresis (PAGE) and size exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS) approaches to determine weight-average molecular weight (M (w)) and polydispersity (PD) of heparins. A set of unfractionated heparin sodium (UFH) and low-molecular-weight heparin (LMWH) samples obtained from nine manufacturers which supply the US market were assessed. For SEC-MALLS, we measured values for water content, refractive index increment (dn/dc), and the second virial coefficient (A (2)) for each sample prior to molecular weight assessment. For UFH, a mean +/- standard deviation value for M (w) of 16,773 +/- 797 was observed with a range of 15,620 to 18,363 (n = 20, run in triplicate). For LMWHs by SEC-MALLS, we measured mean M (w) values for dalteparin, tinzaparin, and enoxaparin of 6,717 +/- 71 (n = 4), 6,670 +/- 417 (n = 3), and 3,959 +/- 145 (n = 3), respectively. PAGE analysis of the same UFH, dalteparin, tinzaparin, and enoxaparin samples showed values of 16,135 +/- 643 (n = 20), 5,845 +/- 45 (n = 4), 6,049 +/- 95 (n = 3), and 4,772 +/- 69 (n = 3), respectively. These orthogonal measurements are the first M (w) results obtained with a large heparin sample set on product being marketed after the heparin crisis of 2008 changed the level of scrutiny of this drug class. In this study, we compare our new data set to samples analyzed over 10 years earlier. In addition, we found that the PAGE analysis of heparinase digested UFH and neat LMWH samples yield characteristic patterns that provide a facile approach for identification and assessment of drug quality and uniformity.