作　　者： (慕艳红）; (徐英）; (侯风萍）; (裴钰）; (傅元欣）; (魏然）; (高雪军）;

机构地区： 兰州生物制品研究所有限责任公司质量检定室甘肃省疫苗工程技术研究中心,甘肃兰州730046

出　　处： 《微生物学免疫学进展》 2017年第4期33-38,共6页

摘　　要： 目的建立检测破伤风抗毒素中甲苯残留量的顶空气相色谱法。方法参照《中国药典》三部(2015版)"残留溶剂测定法",采用Agilent DB-1301毛细管色谱柱(15.0 m×530μm×1.00μm)及氢火焰离子化检测器;顶空进样器各参数分别为:平衡温度70℃,平衡时间30 min,定量环温度80℃,传输线温度90℃,进样量1 m L;气相色谱仪各参数为:进样室温度100℃,柱温箱温度60℃,检测器温度220℃,载气氮气流速5 m L/min,采集时间5min;并对该方法的系统适用性、专属性、线性范围、准确度和精密度、检测限和定量限进行验证及初步应用。结果甲苯保留时间2.429 min,峰面积的RSD为1.9%,以甲苯色谱峰计算得到的理论塔板数N=12 900,甲苯色谱峰与其相邻色谱峰的分离度R=6.03。方法的专属性强,制品中的其他物质不干扰甲苯的出峰;标准曲线的范围为1.0×10-4%~2.0×10-3%,相关系数r大于0.99;加标试验的回收率分别为96.0%~102.9%、100.7%~111.0%,重复性和日间精密度RSD均小于5%。检出限为2.6×10-6%,定量限为1.0×10-5%。测定10批破伤风抗毒素成品和10批破伤风抗毒素原液,其甲苯残留量均低于检出限,远低于规定的限度0.089%。结论该方法简便、快速、准确度、灵敏度高、精密度好,适用于破伤风抗毒素类制品中甲苯残留量的检测。 Objective To set up a method in determination of residual toluene in tetanus antitoxin by using headspace gas chromatography. Methods According to related requirement published in《Chinese Pharmacopoeia 》（2015）,headspace gas chromatography equipped with flame ionization detector and DB-1301（15.0 m× 530 μm× 1.00 μm） column were used.The gas chromatographic conditions were settled as follows： equilibrating temperature 70 ℃,equilibrating time 30 min,inject volume 1 m L,loop temperature 80 ℃,transmission line temperature 90 ℃,injector temperature 100 ℃,column temperature 60 ℃,detector temperature 220 ℃,the flow rate of N25 m L/min,acquisition time 5 min. Then the method was validated including the system suitability test（SST）,the specificity test,the linearity and the range,accuracy and precision,the limit of detection and the limit of quantization. And several batches of tetanus antitoxin were determined by using this method. Results The test showed that retention time of toluene was 2.429 min,RSD of toluene＇s peak areas 1.9%,calculated number of theoretical plates 12 900 and resolution 6.03. The SST met the related requirement. The method was specific to toluene＇s analysis,which indicated that the other components in the preparation did not interfere with formation of toluene peak. The calibration curve was determined in the range of 1.0×10-4%-2.0×10-3%,and the correlation coefficient was 0.99,showed a linear correlation between the content and the area. The recovery of spiked test was 96.0%-102.9% and 100. 7%-111. 0%,respectively,and the RSD of intra and inter day was 5%. The LOD and LOQ was 2.6×10-6and 1.0×10-5,indicating that the method was highly sensitive. In testd10 batches of final products and 10 batches of bulks, t he all detected residual toluene contents were far less than 0. 089% which is specified limitation in requirement. C onclusion This method was rapid,simple,accuracy and highly sensitive. It can be applied in analysis of residual toluene in tetanus antitox

关 键 词： 顶空气相色谱法 甲苯 破伤风抗毒素 验证