目的 建立分子排阻色谱-多角度激光散射-示差折光检测器(SEC-MALLS-RI)联用测定A、C、Y、W135群脑膜炎球菌多糖疫苗(MPV)的相对分子质量与相对分子质量分布的方法。方法 采用TSK Gel G5000 PWxl色谱柱,以0.2 mol·L-1氯化钠为流动相,流速为0.5 mL·min-1,柱温为25 ℃,多角度激光散射器和示差折光检测器联用测定脑膜炎球菌多糖疫苗分子质量,并与2020年版《中国药典》通则3419测定法进行比较。结果 A群、C群、Y群、W135群重均分子质量分别约为20万、30万、15万、30万。结论 SEC-MALLS-RI法可直接测定样品的分子质量与分子质量分布,方法更简便、快速。
Abstract
OBJECTIVE To establish a method for determination of molecular weight and molecular weight distribution of group A, C, Y and W135 meningococcal polysaccharide vaccines by SEC-MALLS-RI. METHODS A gel size exclusion chromatography in conjunction with refractive index detector and multi-angle laser light scattering detector was performed on a TSK Gel G5000 PWxl column, using 0.2 mol·L-1 NaCl solution as mobile phase, the flow rate was 0.5 mL·min-1, and the column temperature was maintained at 25 ℃. And the method was compared with that in General Rule 3419 of ChP 2020. RESULTS The molecular weights of group A, C, Y, W135 were about 200 000, 300 000, 150 000 and 300 000, respectively. CONCLUSION The method is simple and rapid, suitable for the determination of molecular weight and molecular weight distribution of group A, C, Y and W135 meningococcal polysaccharide vaccines.
关键词
脑膜炎球菌多糖疫苗 /
分子排阻色谱 /
多角度激光散射检测器 /
分子质量 /
分子质量分布
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Key words
meningococcal polysaccharide vaccine /
multiangle laser scattering detector /
multi angle laser light scatteringdetector /
molecular weight /
molecular weight distribution
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中图分类号：
R917
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参考文献
[1] Chinese Preventive Medicine Association. Experts′ consensus on immunization with meningococcal vaccines in China[J]. Chin J Epidemiol(中华流行病学杂志), 2019, 40(2):123-128.
[2] ZHANG Y, CAI T, WU Y L, et al. Suitability of Human Whole Blood in Vitro Pyrogen Test for Meningococcal Groups ACYW135 Polysaccharide Vaccine [J]. Chin Pharm J(中国药学杂志), 2015,50(11):983-986.
[3] ZHANG Z, LI X P, YANG F, et al. Progress on capsular polysaccharides and their vaccines [J]. Prog Vet Med (动物医学进展), 2015,36(11):83-87.
[4] Ch.P(2020)Vol Ⅲ(中国药典2020年版. 三部)[S]. 2020:565.
[5] KAN W N, TENG Y K, YANG H W. Determination of molecular weight of Dextran 70 raw material by high performance size exclusion chromatography with differential refraction detector and multi-angle laser light scatter[J]. J China Pharm(中国药房), 2014, 25(9):843-845.
[6] E.P(2015)8.0(欧洲药典8.0版)[S]. 2015:46-47.
[7] E.P(2016)9.0(欧洲药典9.0版)[S]. 2016:47-48.
[8] FAN W W, FAN X D, TIAN W, et al. Differential analysis on precise determination of molecular weight of triblock copolymer using SEC/MALS and MALDI-TOF MS[J]. Polym Testing, 2014,40:116-123.
[9] YANG L X, WANG Z, LI L, et al. Development and verification of SEC-RI-MALS method for determination of molecular weight of group ACYW135 meningococcal polysaccharide-CRM197 conjugate[J]. Chin J Biol (中国生物制品学杂志), 2015, 28(8):841-847.
[10] LUO S Q, ZHAO Z Q, FANG M, et al. Validation of high performance size exclusion chromatographymultiangle laser-light scattering for determination of molecular weight of Haemophilus influenzae type b capsular polysaccharide[J]. Chin J Biol (中国生物制品学杂志), 2015, 28(7):736-740.
[11] KONG S J, YUAN F, WANG X R, et al. Immunogenicity in mice influenced by group C meningococcal conjugates[J]. Prog Microbiol Immunol(微生物学免疫学进展), 2016, 44(1):7-13.
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