National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Anal Bioanal Chem. 2011 Feb;399(4):1449-53. doi: 10.1007/s00216-010-4140-7. Epub 2010 Sep 1.
Size-exclusion chromatography (gel filtration chromatography or gel permeation chromatography) in conjunction with online synchrotron radiation solution small-angle X-ray scattering optics, absorbance, and/or refractive index detectors was further assessed by application of biological macromolecules, such as the hollow sphere protein complex, apoferritin, and a linear polysaccharide, pullulan. The net X-ray scattering patterns of the eluted 24-mer molecule of apoferritin showed the specific character for the hollow spherical shape. The chromatographic (time-resolved) X-ray scattering data of the linear polysaccharide pullulan revealed the flexible chain structure during the chromatographic separation in an aqueous solution. These further applications demonstrated that the present measurement technique will be useful for not only the determination of the radius of gyration value of less than about 10 nm and molecular weight below several hundred thousand but also for the structural characterization of the various macromolecules during the chromatography.
尺寸排阻色谱（凝胶过滤色谱或凝胶渗透色谱）与在线同步辐射溶液小角 X 射线散射光学、吸光度和/或折射率检测器结合使用，进一步应用于生物大分子，如空心球蛋白质复合物、脱铁蛋白和线性多糖普鲁兰。洗脱的脱铁蛋白 24 -mer 分子的净 X 射线散射图谱显示出空心球形的特征。线性多糖普鲁兰的色谱（时间分辨）X 射线散射数据显示了在水溶液中的色谱分离过程中柔性链结构。这些进一步的应用表明，本测量技术不仅将有助于确定小于约 10nm 的回转半径值和几十万以下的分子量，而且还有助于在色谱过程中对各种大分子的结构特征进行分析。
