Molecular weight determination of heparin and dermatan sulfate by size exclusion chromatography with a triple detector array
Molecular weight determination of heparin and dermatan sulfate by size exclusion chromatography with a triple detector array
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DOI：
10.1021/bm049693s
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发表时间：
2005-01-01
影响因子：
6.2
通讯作者：
Terbojevich, M
中科院分区：
文献类型：
--
作者：
Bertini, S;Bisio, A;Terbojevich, M
本文介绍了一种新型的肝素分子质量(M)和分子量分布（MD）的测定方法，该方法由高效粒径排除色谱（HP-SEC）和三重检测器阵列（TDA）组成。HP-SEC/TDA允许通过三个在线检测器，直角激光散射（RALLS），折光计（RI）和粘度计的组合和同时作用来评估聚合物样品的MD。该方法不需要任何色谱柱校准，因此也克服了获得足够参考标准品的困难。它也允许小分子的大小确定，即使散射不对称是不可观察到的。用HP-SEC/TDA分析未分级肝素、8个分级肝素和皮肤聚糖硫酸盐。肝素组分的M值用于建立常规HP-SEC系统的校准曲线：使用HP-SEC/TDA和HP-SEC分析未分离肝素样品的结果非常一致，表明可以使用TDA数据生成具有已知MD和特性粘度的标准样品[eta]。此外，HP-SEC/TDA也可以成功地用于测定Mark-Houwink a和k参数。
The determination of molecular weight (M) and molecular weight distribution (MD) of heparins by a novel approach, consisting of a high performance size exclusion chromatography (HP-SEC) combined with a triple detector array (TDA) is described. HP-SEC/TDA permits the evaluation of MD of polymeric samples through a combined and simultaneous action of three on-line detectors, right-angle laser light scattering (RALLS), refractometer (RI), and viscometer. The method does not require any chromatographic column calibration, thus overcoming also the difficulty to obtain adequate reference standards. It permits the size determination also of small molecules, even when scattering dissimmetry is not observable. Unfractionated heparins, eight fractions of a size fractionated heparin, and dermatan sulfates were analyzed by HP-SEC/TDA. The M values found for the heparin fractions were used to build up a calibration curve of a conventional HP-SEC system: the results obtained analyzing unfractionated heparin samples with both HP-SEC/TDA and HP-SEC were in excellent agreement, suggesting the possibility to use the TDA data to generate standard samples with known MD and intrinsic viscosity [eta]. Moreover, HP-SEC/TDA can successfully be employed also for the determination of the Mark-Houwink a and k parameters.