摘要
目的建立高效液相色谱(high-performance liquid chromatography,HPLC)-电喷雾检测器(charged aerosol detection,CAD)法测定新型冠状病毒mRNA疫苗中的蔗糖含量,并对方法进行验证及初步应用,以期为更多mRNA疫苗中的蔗糖含量测定提供新的检测方法。方法色谱柱:Zorbax NH2(Agilent,4.6 mm×250 mm,5.0μm);色谱条件:以乙腈溶于水(乙腈50%)为流动相,流速0.8 mL/min,雾化温度70℃,检测器采集频率10 Hz,滤波3.6 s,柱温20℃。对方法的线性、定量限与检出限、专属性、重复性、准确性进行验证,并采用建立的方法检测3批新型冠状病毒mRNA疫苗中的蔗糖含量。结果蔗糖标准品浓度在0.25~6 mg/mL范围内与峰面积线性关系良好,相关系数(R)为0.9986;该方法检出限和定量限分别为0.03125和0.125 mg/mL;乳糖和蔗糖在本试验条件下能够完全分开,分离度为1.65;6份供试品峰面积的相对标准偏差(relative standard deviation,RSD)为2.4%;3个不同浓度的加标溶液回收率为101.3%~102.2%。采用建立的方法检测3批新型冠状病毒mRNA疫苗样品的蔗糖含量分别为81.3774、80.3390和74.1573 mg/mL。结论建立的HPLC-CAD方法测定范围宽,线性和重复性好,准确性高,能够对新型冠状病毒mRNA疫苗中的蔗糖进行准确定量。
Objective To establish a high-performance liquid chromatography(HPLC)with charged aerosol detection(CAD)method for the determination of sucrose content in mRNA-based SARS-CoV-2 vaccines,and to validate and preliminarily apply the method,so as to provide a new detection method for the determination of sucrose content in more mRNA vaccines.Methods Chromatographic column:Zorbax NH_2(Agilent,4.6 mm×250 mm,5.0μm);Chromatographic conditions:acetonitrile dissolved in water(50%acetonitrile)as the mobile phase,the flow rate of 0.8 mL/min,the nebulization temperature of 70℃,the detector acquisition frequency of 10 Hz,the filtration of 3.6 s,and the column temperature of 20℃.The linearity,limit of quantification(LOQ)and limit of detection(LOD),specificity,repeatability and accuracy of the method were validated,and the sucrose content in three batches of mRNA-based SARS-CoV-2 vaccines was detected by the established method.Results The concentration of the sucrose standard exhibited a good linear correlation with peak area in R therangeof0.25 to6 mg/mL,withacorrelationcoefficient()of0.9986.TheLODandLOQwere0.03125 and0.125 mg/mL respectively.Lactose and sucrose could be completely separated under the experimental conditions,and the resolution was1.65.The relative standard deviation(RSD)of peak area of the six test samples was 2.4%.The recovery rates for spiked samples at three different concentrations ranged from 101.3%to 102.2%.The sucrose content of three batches of SARS-CoV-2 mRNA vaccine samples detected by the established method was 81.3774,80.3390 and 74.1573 mg/mL,respectively.Conclusion The established HPLC-CAD method has a wide measurement range,good linearity and repeatability,and high accuracy,allowing for accurate quantification of sucrose content in the mRNA-based SARS-CoV-2 vaccines.
作者
吴凡
刘勤
杨欢
申雪
王思捷
和文志
非成瑞
WU Fan;LIU Qin;YANG Huan;SHEN Xue;WANG Sijie;HE Wenzhi;FEI Chengrui(Yunnan Institute of Supervision and Inspection for Food and Drug,Public Service Platform for Industrial Technology Foundation of the Ministry of Industry and Information Technology,Kunming 650106,Yunnan Province,China)
出处
《中国生物制品学杂志》
2025年第11期1351-1357,共7页
Chinese Journal of Biologicals
基金
云南省科技厅重大科技专项计划(202002AA100008)。
关键词
蔗糖含量
高效液相色谱法
电喷雾检测器
新型冠状病毒mRNA疫苗
Sucrose content
High-performance liquid chromatography(HPLC)
Charged aerosol detection(CAD)
mRNA-based SARS-CoV-2 vaccines
