不同方法对寡聚甘露糖醛酸铬配合物分子量测定的影响  

Influence of different methods on molecular mass determination of oligomannuronate-chromium(Ⅲ)complex

作　　者：李广生[1] 赵峡[1] 郝翠[1] 吕友晶[1] 曾洋洋[1] 于广利[1] 

机构地区：[1]中国海洋大学海洋药物教育部重点实验室,山东省糖科学与糖工程重点实验室,山东青岛266003

出　　处：《中国海洋药物》2014年第1期6-10,共5页Chinese Journal of Marine Drugs

基　　金：国家科技支撑计划(2013BAB01B02);国家高技术研究发展计划(2007AA09Z445);国家自然科学基金(31070724);海洋局公益专项(201005024)资助

摘　　要：目的探讨不同方法对于测定具有抗2型糖尿病活性的海洋寡聚甘露糖醛酸铬配合物（HS203）分子量测定的影响。方法采用聚丙烯酰胺凝胶电泳（PAGE），高效凝胶渗透色谱（HPGPC）及高效凝胶渗透色谱与多角度激光光散射器联用（HPGPC-MALLS）法分别测定HS203的分子量，并考察了不同色谱柱以及不同分子量对照品的选择对于HS203分子量测定结果的影响。结果PAGE法表明HS203的聚合度为5～20之间，通过计算得出HS203的重均分子量范围在2400-2600Da。在HPGPC分析过程中，不同色谱柱对于HS203分子量测定结果影响不大，采用右旋糖酐作为对照品测得分子量为6000-7100Da，明显大于PAGE结果，采用结构相似的自制对照品测定结果为2900～3100Da，与PAGE结果较为接近。采用HPGPC-MALLS法测定结果约为3000Da，也与PAGE法所测结果相近。结论不同测定方法测得的HS203分子量结果不同。PAGE法能直观地观察到HS203聚合度的分布情况，并能真实反映分子量大小。HPGPC法应用普遍，操作方面，但对照品的选择对HPGPC法测定分子量十分重要，必须选用与待测样品结构相似的对照品。HPGPC-MALLS法测定糖类化合物分子量无需对照品且结果可靠，但该法需要专用且昂贵的仪器。Objective To investigate different methods for the determination of molecular mass （Mr） of marine oligomannuronate-chromium （Ⅲ） complex（HS23）which has anti-diabetes Ⅱ activity. Methods Polyacrylamide gel electrophoresis（PAGE）, high performance gel permeation chromatography（HPGPC） and high performance gel permeation chromatography combined with multi-angle laser light scattering （HPGPC-MALLS）were used to determine the molecular mass of HS203 and compare the influence on Mr determination by HPGPC with different columns and standards. Results PAGE results showed that the degree of polymerization of HS203 was between 5-20 and the weight average molecular mass was estimated between 2 400-2 600 Da. HPGPC results showed that the Mr of HS203 determined by different column was similar, and the Mr was between 6 000~7 100 Da using dextran as standards, the result was significantly larger than that by the PAGE method. But the Mr was between 2 900-3 100 Da by using the standards having similar structure with HS203, which was close to the PAGE result. The Mr determined by HPGPC-MALLS was 3 000 Da, which was also similar with PAGE result. Conclusion The Mr of HS203 determined by different methods was not the same. PAGE showed the degree of polymerization, its real distribution and Mr of HS203. HPGPC was widely used and more convenient, but the choice of different standards apparently effected the Mr result and the standards＇ structures should be similar to the samples for obtaining the accurate and reliable results. The molecular mass of carbohydrate could be determined directly and accurately without standards by HPGPC-MALLS method, how- ever, it required specific and expensive equipments.

关 键 词：寡聚藻甘酸铬配合物 聚丙烯酰胺凝胶电泳 高效凝胶渗透色谱 多角度激光光散射 分子量 

分 类 号：O629.12[理学—有机化学] R282.77[理学—化学]