SEC-RI-MALLS测定甘露聚糖肽分子量与分子量分布
Determination of molecular weight and distribution of mannatide by means of SEC-RI-MALLS
邹剑 1杨蕾 1林涛 1王觉晓 1马晶 1陈婕 1李炎1
作者信息
- 1. 四川省药品检验研究院国家药品监督管理局疫苗及生物制品质量控制与评价重点实验室四川省药品监督管理局生物制品质量监测与风险评估重点实验室,成都 610097
- 折叠
摘要
目的:建立尺寸排阻色谱-示差折光-多角度激光光散射检测器联用(size exclusion chromatography-refractive index-multiangle laser light scattering,SEC-RI-MALLS)测定甘露聚糖肽原料药和制剂分子量及分子量分布的方法.方法:采用SEC-RI-MALLS联用技术,以0.05 mol·L-1硫酸钠溶液为流动相,Shodex OHpak SB-804 HQ为色谱分离柱,在流速0.5 mL·min-1,柱温35℃的条件下对甘露聚糖肽分子量及其分布进行专属性、准确度、精密度、耐用性方法学验证,并与现行质量标准方法进行比较.结果:片剂辅料淀粉对测定无干扰,右旋糖酐标准品实测值同标示值的准确度相对误差＜3.0％;当样品配制浓度为2 mg·mL-1时精密度最好,RSD为0.40％;重现性、耐用性RSD均不大于5.0％.79批次样品采用Shodex OHpak SB-804 HQ色谱柱和TSK-GEL G4000 PWXL色谱柱进行测定,结果无显著性差异.SEC-RI-MALLS法比甘露聚糖肽国家标准方法(GPC法)的结果平均高19 509 Da,且分子量分布更为集中.结论:SEC-RI-MALLS法无需标准品就能够测定甘露聚糖肽分子量及其分布,结果准确,耐用性好,与现行方法相比,更有利于该品种的安全性、有效性控制.
Abstract
Objective:To establish a method for determining molecular weight and distribution of raw mannatide and its preparation with SEC-RI-MALLS.Methods:Specificity,accuracy,precision and robustness of SEC-RI-MALLS method was verified by performing on a Shodex OHpak SB-804 HQ column with 0.05 mol·L-1 so-dium sulfate buffer as mobile phase at a flow rate of 0.5 mL·min-1 and a comparison between SEC-RI-MALLS and GPC was studied too.Resulst:The tablet excipient starch had no interference to the test.The rela-tive accuracy error between the measured value and the labeled value of dextran standard was less than 3.0％.RSD of precision was 0.40％when the sample concentration was 2 mg·mL-1 and the RSD of reproducibility and robustness were less than 5.0％.There was no significant difference between Shodex OHpak SB-804 HQ column and TSK-GEL G4000 PWXL column by comparing the results of 79 batches of samples.Compared with the national standard method(GPC)for molecular weight determination,the molecular weight of SEC-RI-MALLS method was 19 509 Da higher on average,and the molecular weight distribution was more concentrated.Conclusion:SEC-RI-MALLS method can determine the molecular weight and distribution of mannatide without standard with good accuracy and robustness.Compared with the existing method,SEC-RI-MALLS method is more conducive to the safety and effectiveness control of the variety.
关键词
多角度激光光散射/尺寸排阻色谱/凝胶渗透色谱/甘露聚糖肽/分子量与分子量分布Key words
multi-angle laser light scattering/size exclusion chromatography/gel permeation chromatography/mannatide/molecular weight and distribution引用本文复制引用
出版年
2024
NETL
NSTL
万方数据