Hokputsa Sanya, Jumel Kornelia, Alexander Catherine, Harding Stephen E
NCMH Physical Biochemistry Laboratory, School of Biosciences, University of Nottingham, Sutton Bonington, LE12 5RD, UK.
Eur Biophys J. 2003 Aug;32(5):450-6. doi: 10.1007/s00249-003-0299-6. Epub 2003 Apr 16.
Hyaluronic acid (HA) is a natural polysaccharide with importance in the pharmaceutical, medical and cosmetic industries. Determining factors in its final applications are its physicochemical properties, particularly molecular mass. A high molecular mass HA was degraded using five different hydroxyl free-radical starting concentrations chemically produced from ascorbic acid and hydrogen peroxide. The aims of the study were to investigate the effect of different hydroxyl free-radical concentrations on the chain length of HA and compare the molecular masses obtained from analytical ultracentrifugation using sedimentation equilibrium experiments and size exclusion chromatography/multi-angle laser light scattering (SEC/MALLS). The results indicated that their molecular masses varied, depending on the degree of hydroxyl free-radical starting concentration. Molecular mass values obtained from sedimentation equilibrium experiments for each sample showed the same trend as those obtained from the SEC/MALLS in the range of molecular masses studied. The molecular masses obtained from sedimentation equilibrium for high molecular mass samples from reciprocal plots of apparent weight average molecular mass against concentration gave values similar to those obtained by SEC/MALLS. In contrast, the molecular mass from conventional plots for high molecular mass samples were much lower than those from SEC/MALLS, even when high ionic strength buffers were used.
透明质酸（HA）是一种天然多糖，在制药、医学和化妆品行业具有重要意义。其最终应用中的决定性因素是其物理化学性质，尤其是分子量。使用由抗坏血酸和过氧化氢化学产生的五种不同起始浓度的羟基自由基对高分子量HA进行降解。本研究的目的是研究不同羟基自由基浓度对HA链长的影响，并比较通过沉降平衡实验的分析超速离心法和尺寸排阻色谱/多角度激光光散射（SEC/MALLS）获得的分子量。结果表明，它们的分子量随羟基自由基起始浓度的程度而变化。在研究的分子量范围内，每个样品通过沉降平衡实验获得的分子量值与通过SEC/MALLS获得的分子量值显示出相同的趋势。从表观重均分子量对浓度的倒数图中获得的高分子量样品沉降平衡的分子量值与通过SEC/MALLS获得的值相似。相比之下，即使使用高离子强度缓冲液，高分子量样品常规图的分子量也远低于SEC/MALLS的分子量。
