AEX-HPLC法测定单纯疱疹病毒注射液中病毒颗粒数  

Determination of viral particle in herpes simplex virus injection by AEX-HPLC

作　　者：孙名皓 雷雨 胡国栋 SUN Minghao;LEI Yu;HU Guodong(Shanghai Pharma Sunway Biotech Co.,Ltd.,Shanghai 201206,China)

机构地区：[1]上海三维生物技术有限公司,上海201206

出　　处：《上海医药》2024年第15期83-86,共4页Shanghai Medical & Pharmaceutical Journal

摘　　要：目的:建立检测单纯疱疹病毒注射液病毒颗粒数(VP)的阴离子交换色谱法(AEX-HPLC)。方法:采用Resource Q分析柱和Agilent 1260Ⅱ高效液相色谱系统,以Tris缓冲溶液(20 mmol/L,pH 7.0)为流动相A和以含2.5 mol/L氯化钠的Tris缓冲溶液为流动相B进行梯度洗脱,并对方法进行验证。结果:空白溶剂无干扰峰出现;未经纯化的单纯疱疹病毒溶液中的疱疹病毒峰与其他杂质峰分离度大于1.5;该方法的定量限和检测限为5.0×10^(7)VP/mL和2.0×10^(7)VP/mL;疱疹病毒的进样量在5×10^(7)~1×10^(9)病毒颗粒范围内,浓度与吸收峰的线性关系良好(R^(2)=0.9982);整个精密度实验的RSD%为0.9%;回收率70%~130%之间。结论:AEX-HPLC法可用于检测单纯疱疹病毒注射液的病毒颗粒数。Objective:To establish an assay method for the determination of viral particle(VP)in herpes simplex virus injection by AEX-HPLC.Methods:The HPLC was run on Resource Q column with gradient elution using Tris buffer(20 mmol/L,pH 7.0)as mobile phase A and Tris buffer containing 2.5 mol/L NaCl as mobile phase B.The assay method was subsequently validated.Results:There was no interference peak in the blank solvent and the resolution between peaks for herpes simplex virus and other impurities in the unpurified herpes simplex virus solution was greater than 1.5.The LOQ and the LOD of this assay were 5.0×10^(7) and 2.0×10^(7) VP/mL.The linearity relationship between the concentration and the absorption peak was good in the range of 5×10^(7)-1×10^(9) VP/mL with R20.9982.RSD%in precision assay was 0.9%and recovery in accuracy assay was between 70%-130%.Conclusion:The AEX-HPLC assay can be applied to determine the viral particle in herpes simplex virus injection.

关 键 词：单纯疱疹病毒 病毒颗粒 分析方法验证 

分 类 号：O657.75[理学—分析化学] R927.2[理学—化学]