Hennink W E, van den Berg J W, Feijen J
Thromb Res. 1987 Mar 1;45(5):463-75. doi: 10.1016/0049-3848(87)90309-4.
This study shows that HPLC-LALLS (high performance liquid chromatography with a light-scattering detector) is a convenient and reliable method for the characterization of standard heparin samples, provided that polyelectrolyte artefacts are suppressed by a suitable dialysis procedure. The method has been successfully applied to the assessment of calibrated relations like the Mark-Houwink relation and the calibration curve of gel permeation chromatography. Furthermore, this study confirms that gel permeation chromatography for heparins cannot be calibrated with dextranes or polyethyleneglycols, but in addition to this well known fact it is shown that even the universal (hydrodynamic volume) calibration procedure does not work. Moreover, in addition to the familiar conclusion that heparins should be calibrated with heparins, it is shown that calibrated relations are valid only when the calibration has been made with heparin samples of the same origin (i.e. same tissue source, same animal source, and same purification scheme).
本研究表明，高效液相色谱-激光光散射检测法（HPLC-LALLS）是表征标准肝素样品的一种便捷可靠的方法，前提是通过合适的透析程序抑制聚电解质假象。该方法已成功应用于评估校准关系，如马克-豪温克关系和凝胶渗透色谱的校准曲线。此外，本研究证实，肝素的凝胶渗透色谱不能用葡聚糖或聚乙二醇校准，而且除了这一众所周知的事实外，还表明即使是通用（流体动力学体积）校准程序也不起作用。此外，除了肝素应用肝素进行校准这一常见结论外，还表明校准关系仅在校准是使用相同来源（即相同组织来源、相同动物来源和相同纯化方案）的肝素样品进行时才有效。
