PRACTICAL ONLINE DETERMINATION OF BIOPOLYMER MOLECULAR-WEIGHTS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH CLASSICAL LIGHT-SCATTERING DETECTION
PRACTICAL ONLINE DETERMINATION OF BIOPOLYMER MOLECULAR-WEIGHTS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH CLASSICAL LIGHT-SCATTERING DETECTION
复制标题
DOI：
10.1016/0021-9673(92)85088-b
复制
发表时间：
1992-02-21
期刊：
影响因子：
--
通讯作者：
JONES, R
中科院分区：
文献类型：
--
作者：
DOLLINGER, G;CUNICO, B;JONES, R
大多数生物聚合物分子比经典光散射实验中使用的光的波长小得多。500 nm），因此简单的瑞利方程和90度光散射光度计足以确定它们的分子量。结合高效液相色谱（HPLC），它表明，一个简单的HPLC荧光计可以用作90度光散射检测器的生物聚合物分子量测定。为了简化数据处理，仅测量相对分子量。采用了三个数学假设，并证明了它们对蛋白质的有效性。为了将工作的角度来看，这种90度光散射检测器的相对优势和局限性进行比较，更常用的低角度激光光散射检测器。蛋白质分子量测定的两个例子来说明90度经典光散射的生物聚合物的结构和相互作用的研究的广泛用途。
Most biopolymer molecules are much smaller than the wavelength of light used in classical light-scattering experiments (ca. 500 nm), and thus the simple Rayleigh equation and a 90-degrees light-scattering photometer are sufficient to determine their molecular weight. In combination with high-performance liquid chromatography (HPLC), it is demonstrated that a simple HPLC fluorimeter can be used as a 90-degrees light-scattering detector for biopolymer molecular weight determinations. To simplify data handling, only relative molecular weights are measured. Three mathematical assumptions are adopted, and their validity for proteins is shown. To place the work in perspective, the relative advantages and limitations of this 90-degrees light-scattering detector are compared with the more commonly used low-angle laser light-scattering detector. Two examples of protein molecular weight determinations are given to illustrate the broad utility of 90-degrees classical light scattering to the study of biopolymer structures and interactions.