Edens R E, al-Hakim A, Weiler J M, Rethwisch D G, Fareed J, Linhardt R J
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
J Pharm Sci. 1992 Aug;81(8):823-7. doi: 10.1002/jps.2600810821.
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.
作为开发药物肝素和低分子量（LMW）肝素衍生物表征方法的合作研究的一部分，对其分子量（M(r)）值进行了检测。分子量测定的标准方法依赖于凝胶渗透高效液相色谱法（HPLC）。我们报告了使用梯度聚丙烯酰胺凝胶电泳（PAGE）来测定药物肝素和LMW肝素衍生物的M(r)值。这种方法相对于HPLC方法具有某些优势。梯度PAGE分析在多个样品上并行进行，使用相同的标准曲线。HPLC则是串行进行。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和传统的凝胶渗透HPLC方法对肝素和各种LMW肝素样品进行了分析。两种技术都对数均分子量（M(r)）、重均分子量（M(r)）和多分散性进行了检测，结果发现它们相似。这项研究表明，梯度PAGE分析是测定肝素和LMW肝素M(r)值的一种灵敏方法。
