GRADIENT POLYACRYLAMIDE-GEL ELECTROPHORESIS FOR DETERMINATION OF MOLECULAR-WEIGHTS OF HEPARIN PREPARATIONS AND LOW-MOLECULAR-WEIGHT HEPARIN DERIVATIVES
GRADIENT POLYACRYLAMIDE-GEL ELECTROPHORESIS FOR DETERMINATION OF MOLECULAR-WEIGHTS OF HEPARIN PREPARATIONS AND LOW-MOLECULAR-WEIGHT HEPARIN DERIVATIVES
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DOI：
10.1002/jps.2600810821
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发表时间：
1992-08-01
影响因子：
3.8
通讯作者：
LINHARDT, RJ
中科院分区：
文献类型：
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作者：
EDENS, RE;ALHAKIM, A;LINHARDT, RJ
作为合作研究的一部分，对药物肝素和低分子量 (LMW) 肝素衍生物的 M(r) 值进行了检查，以开发其表征方法。 M(r) 测定的标准方法依赖于凝胶渗透高效液相色谱 (HPLC)。我们报告了使用梯度聚丙烯酰胺凝胶电泳 (PAGE) 来确定药物肝素和 LMW 肝素衍生物的 M(r) 值。与 HPLC 方法相比，该方法具有一定的优势。使用相同的标准曲线对多个样品平行进行梯度 PAGE 分析。连续进行HPLC。梯度 PAGE 比 HPLC 具有更高的分辨率，因此，使用易于获得的标准品混合物代替单个标准品来构建标准曲线。通过梯度 PAGE 和常规凝胶渗透 HPLC 方法对肝素和各种 LMW 肝素样品进行分析。通过这两种技术检查了数均 M(r)、重均 M(r) 和多分散性，发现它们相似。本研究表明梯度 PAGE 分析是测定肝素和 LMW 肝素 M(r) 值的灵敏方法。
The M(r) values of pharmaceutical heparins and low-molecular-weight (LMW) heparin derivatives were examined as part of a collaborative study to develop methods for their characterization. Standard methods of M(r) determination rely on gel permeation high-performance liquid chromatography (HPLC). We report the use of gradient polyacrylamide gel electrophoresis (PAGE) to determine the M(r) values of pharmaceutical heparins and LMW heparin derivatives. This approach offers certain advantages over the HPLC method. Gradient PAGE analysis was performed in parallel, on multiple samples, with the same standard curve. HPLC was performed serially. Gradient PAGE gave higher resolution than HPLC, and thus, a mixture of easily obtained standards was used in place of individual standards for the construction of a standard curve. Heparin and various LMW heparin samples were analyzed by both gradient PAGE and conventional gel permeation HPLC methods. The number-average M(r), weight-average M(r), and polydispersity were examined by both techniques and found to be similar. This study demonstrates that gradient PAGE analysis is a sensitive method for the determination of the M(r) values of heparin and LMW heparin.