多角度激光光散射仪与尺寸排阻色谱法联用测定透明质酸相对分子质量及其分布

Determination of Molecular Weight and Its Distribution of Hyaluronic Acid by Combination of Multi-angle Laser Light Scattering Instrument and Size Exclusion Chromatography

目的多角度激光光散射仪与体积排阻色谱法联用(MALLS-SEC)测定透明质酸钠(HA)相对分子质量(Mr)及其分布。方法考察MALLS-SEC法测定HA Mr的准确度和精密度,以及HA溶液浓度对测定的影响;比较此法、黏度法和尺寸排阻色谱法测定HA Mr的结果。结果MALLS-SEC法的准确度、精密度均较好,相对标准偏差小于2%;测定Mr为104以上的HA样品采用0.05 mg/mL的浓度较为适宜;MALLS-SEC法测定HAMr的数值与黏度法较为接近,体积排阻色谱法测定值偏高。结论MALLS-SEC法测定HA Mr,操作简单快速,结果准确,可用于HA的质量控制和应用研究。

Objective To determine the molecular weight （M） and its distribution of hyaluronic acid （HA） by the combination of multi-angle laser light scattering instrument and size exclusion chromatography （MALLS-SEC）. Methods The accuracy and precision were studied in the determination of M and distribution of HA by MALLS- SEC. The effect of concentration of HA solution on the determination of molecular weight was investigated. The determination results of M were compared from MALLS-SEC, SEC, viscosity methods. Results The MALLS-SEC method showed high accuracy and good precision with RSD〈2 %. The HA solution of 0.05 mg/mL was more suitable for the determination of M more than 104 by MALLS-SEC. The M of HA determined by MALLA-SEC method was close to that measured by viscosity method. However, the Mr determined by SEC method was too high. Conclusion The MALLS-SEC method is simple, fast and accurate for the determination of Mr and distribution of HA, and it can be used for the quality control of HA