反相高效液相色谱法检测吸附无细胞百白破联合疫苗2-苯氧乙醇含量  

Determination of 2-phenoxyethanol content in acellular diphtheria,tetanus and pertussis combined vaccine by reverse phase high performance liquid chromatography

作　　者：邓海清[1] 尹珊珊[1] 刘杨[1] 张艳红[1] 郝倩[1] 高宇皎 任玉莹[1] 殷建齐 邢盛宇 刘建凯[1] DENG Hai-qing;YIN Shan-shan;LIU Yang;ZHANG Yan-hong;HAO Qian;GAO Yu-jiao;REN Yu-ying;YIN Jian-qi;XING Sheng-yu;LIU Jian-kai(Center of Research and Development,Beijing Minhai Biotechnology Co.,Ltd.,Beijing 102600,China)

机构地区：[1]北京民海生物科技有限公司研发中心,北京102600

出　　处：《中国生物制品学杂志》2018年第9期990-994,共5页Chinese Journal of Biologicals

摘　　要：目的建立检测吸附无细胞百白破联合疫苗(diphtheria,tetanus and acellular pertussis combined vaccine,DTa P)中2-苯氧乙醇含量的反相高效液相色谱法(RP-HPLC)。方法采用ZORBAX Eclipse XDB-C18色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈∶水(1∶1),流速1.0 m L/min,检测波长270 nm,柱温25℃,以外标法测定2-苯氧乙醇含量,并对该方法进行线性、重复性、中间精密度、准确度、专属性、耐用性验证。结果 2-苯氧乙醇浓度在100~500μg/m L范围内,与峰面积线性关系良好,R2大于0.99;3批供试品重复性试验2-苯氧乙醇含量相对标准偏差(RSD)分别为0.8%、0.4%和1.2%,中间精密度验证2-苯氧乙醇含量RSD分别为2.38%、2.36%和2.68%;准确度回收率在97.4%~101.9%之间;供试品中加入戊二醛和甲醛对2-苯氧乙醇含量的检测无干扰,专属性较好;温度变化对检测结果无显著影响,耐用性较好。结论该检测方法简单、快速、可靠,可用于疫苗中2-苯氧乙醇含量检测。Objective To develop a reverse phase high performance liquid chromatography（RP-HPLC） method for determination of the 2-phenoxyethanol content in diphtheria,tetanus and acellular pertussis combined vaccine（DTa P）.Methods The 2-phenoxyethanol content was determined with an external standard by HPLC on a ZORBAX Eclipse XDBC18 column（4. 6 mm × 250 mm,5 μm）,serving acetonitrile ∶ water（1 ∶ 1）as mobile phase at a flow rate of 1. 0 m L/min,a detection wavelength of 270 nm and a column temperature of 25 ℃. This method was verified for linearity,reproducibility,intermediate precision,accuracy,specificity and durability. Results The 2-phenoxyethanol content at a range of 100 ~ 500 μg/m L showed good linear relationship to the peak area,with a R2 value of more than 0. 99. The relative standard deviations（RSDs）of 2-phenoxyethanol contents in three batches of samples in reproducibility test were 0. 8%,0. 4% and 1. 2%,while those in intermediate precision test were 2. 38%,2. 36% and 2. 68%,respectively. The recovery rate in accuracy test was 97. 4% ~ 101. 9%. The addition of glutaraldehyde or formaldehyde showed no interference to the determination of 2-phenoxyethanol content, indicating a high specificity. However, temperature showed no significant effect on the determination result,indicating a good durability of the method. Conclusion The method is simple,rapid and reliable,which may be used for determination of 2-phenoxyethanol content in vaccines.

关 键 词：反相高效液相色谱法 吸附无细胞百白破联合疫苗 2-苯氧乙醇 含量测定 

分 类 号：O657.72[理学—分析化学] R392-33[理学—化学]