Komatsu H, Yoshii K, Ishimitsu S, Okada S, Takahata T
Division of Drugs, National Institute of Hygienic Sciences, Osaka, Japan.
J Chromatogr. 1993 Jul 30;644(1):17-24. doi: 10.1016/0021-9673(93)80115-o.
A high-performance gel permeation chromatographic system with on-line low-angle laser light-scattering detection (HPGPC-LALLS) was used to determine molecular masses of low-molecular-mass heparins (LMMHs). Measurements at wide and narrow collection angles were compared and the application of the HPGPC-LALLS method to small molecules, with molecular masses in the range 1000-10,000, was assessed. The molecular mass averages of fractionated heparins and commercially available LMMHs were also determined by ordinary HPGPC analysis using an LMMH molecular-mass calibrant, supplied by the National Institute for Biological Standards and Control for the calibration of columns. The LALLS intensity at the routinely used narrow collection angle (theta col = 1 degree) about doubled at the wide collection angle (theta col = 2 degrees) and the signal-to-noise ratio was improved. The present study thus indicates that wide collection angle measurement of light scattering allows the application of the HPGPC-LALLS method to very small biopolymers of molecular mass < 10,000.
采用带有在线低角度激光光散射检测的高效凝胶渗透色谱系统（HPGPC-LALLS）测定低分子量肝素（LMMH）的分子量。比较了宽收集角和窄收集角下的测量结果，并评估了HPGPC-LALLS方法对分子量在1000 - 10000范围内的小分子的应用。还通过使用由国家生物标准与控制研究所提供的用于柱校准的LMMH分子量校准物进行普通HPGPC分析，测定了分级肝素和市售LMMH的分子量平均值。在常规使用的窄收集角（θcol = 1°）下的LALLS强度在宽收集角（θcol = 2°）时约增加一倍，且信噪比得到改善。因此，本研究表明，光散射的宽收集角测量使得HPGPC-LALLS方法能够应用于分子量<10000的非常小的生物聚合物。
