玻璃酸钠及制剂分子质量与分子质量分布测定方法研究

目的：分析玻璃酸钠（SH）及制剂分子质量和分子质量分布测定不同模式的特点，重点研究折光指数增量（dn/dc值）、对照品等对测定结果的影响。方法：采用凝胶色谱法（SEC），色谱柱为ShodexSB806HQ（300 mm×8 mm），流动相为0.2 mol·L^-1的氯化钠溶液（含0.02%叠氮化钠），流速为0.5 mL·min^-1，柱温为35 ℃，进样量100 μL，检测采用2种方法，单一示差折光检测器及采用示差折光检测器和激光光散射仪（MALLS）联用。结果：玻璃酸钠及制剂相对分子质量和绝对分子质量测定结果存在约30%的误差，dn/dc值、对照品等对测定结果产生一定影响。结论：凝胶色谱示差折光检测法和凝胶色谱激光光散射联用法，2种方法均可用于玻璃酸钠及制剂分子质量控制。

Objective: To analyze the different determination modes of molecular weight and molecular weight distribution of sodium hyaluronate(SH) and its preparations,especially the influence of dn/dc value and standards on the determination results. Methods: Using SEC method,samples were analyzed on a Shodex SB806HQ column(300 mm×8 mm);the mobile phase was 0.2 mol·L^-1 sodium chloride(containing 0.02% sodium azide) at a flow rate of 0.5 mL·min^-1;the injection was 100 μL;the column temperature was 35 ℃,and the signals were detected by RI detector (used by SEC method) or RI coupled with MALLS detectors(used by SEC-MALLS method). Results: The results showed that there were about 30% differences between the molecular weight and molecular weight distribution results determined by SEC-RI and SEC-MALLS methods,and the dn/dc value and standards had certain influences on the determination results. Conclusion: SEC-RI and SEC-MALLS methods could both be used for quality control of SH and its preparations.

[1] YANG Xiao-hong(杨晓红), LING Pei-xue(凌沛学), WANG Feng-shan(王凤山).The developing progress of sodium hyaluronate in clinical application(透明质酸预防外科术后粘连的研究进展)[J].Chin J Biochem Pharm(中国生化药物杂志), 1998, 19(4):205
[2] WANG Yan-hou(王彦厚), WANG Feng-shan(王凤山), GUO Xue-ping(郭学平).Comparison of three relative molecular weight determination methods for low molecular weight hyaluronic acid(测定低分子量透明质酸相对分子量的3种方法比较)[J].Chin J Biochem Pharm(中国生化药物杂志), 2007, 28(3):182
[3] LIU Hui(刘辉), RAO Fang-ping(饶放萍), ZHONG Xue-ping(钟雪平), et al.Study on the effect of sodium hyaluronate on preventing the knee joint adhesion after fracture operation(透明质酸钠在骨折手术后膝关节粘连预防中的效果研究)[J]. Med Inovat China (中国医学创新), 2013, 10(32):49
[4] SONG Yong-min(宋永民), DING Hou-qiang(丁厚强), LUAN Yi-hong(栾贻宏), et al.Study on lubricity of hyaluronan medical lubricant(透明质酸医用润滑剂的润滑性研究)[J].J Biomed Eng Res(生物医学工程研究), 2013, 28(3):182
[5] WANG Hai-ying(王海英), GUO Xue-ping(郭学平), LUAN Yi-hong(栾贻宏).Determination of relative molecular mass and its distribution of sodium hyaluronate by size exclusion chromatography(分子排阻色谱法测定玻璃酸钠相对分子量及其分布)[J].Food Drug(食品与药品), 2010, 12(7):232
[6] WANG Yan-hou(王彦厚), WANG Feng-shan(王凤山), GUO Xue-ping(郭学平).Studies on the determination method of intrinsic viscosity of hyaluronate sodium(玻璃酸钠特性黏度测定方法的研究)[J].Chin Pham J(中国药学杂志), 2004, 39(6):469
[7] AI-Assaf S, Williams PA, Phillips GO.Molecular characterization of hyaluronan and hylan using GPC-MALLS and asymmetrical flow FFF-MALLS[J].Hyaluronan, 2002, 1:55
[8] EP 7.0[S].2010:2927
[9] JP 16.Supplement[S].2011:1056
[10] SONG Yu-juan(宋玉娟) HAN Chun-xia(韩春霞), FAN Hui-hong(范慧红).Determination of molecular weight and molecular weight distribution of sodium hyaluronate(玻璃酸钠分子量与分子量分布测定)[J].Chin J Pharm Anal(药物分析杂志), 2010, 30(12):2430
[11] LIU Li-li(刘莉莉), PAN Hua-xian(潘华先), SHI Yan-ping(施燕平), et al. Study on determination of absolute molecular weight and molecular weight distribution in medical sodium hyaluronate gel(医用透明质酸钠凝胶的绝对分子量及其分布的测定方法研究)[J].Chin J Pharm Anal(药物分析杂志), 2013, 33(8):1435
[12] American Polymer Standards Corporation.Light Scattering dn/dc Values[EB/OL].[2013-05-15].http://www.ampolymer.com/dndc.html