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目的建立亚单位流感疫苗中裂解剂壬苯醇醚-9含量的HPLC检测方法。方法采用4倍体积甲醇沉淀处理亚单位流感疫苗样品,HPLC色谱条件为:色谱柱:Thermo BioBasic-4 C4柱(4.6 mm×150 mm,5μm),流动相:甲醇∶水=80∶20(v/v),流速:1 ml/min,紫外检测器检测波长:275 nm,柱温:33℃,上样量:100μl。并对建立的HPLC法进行验证。结果建立的HPLC法检测壬苯醇醚-9标准品在5~2 500μg/ml浓度范围内线性关系良好;检测壬苯醇醚-9的专属性良好;检测壬苯醇醚-9在灭活流感全病毒提取液和亚单位流感疫苗中的回收率约为100%;检测各浓度壬苯醇醚-9标准品溶液和亚单位流感疫苗的日内和日间精密性均小于3%;检测限和定量限均为5μg/ml。结论建立的HPLC法简便、快速、准确,适用于实验室对亚单位流感疫苗中裂解剂含量的常规检测。
Objective To establish a HPLC method for the determination of nonoxynol-9 in subunit influenza vaccine. Methods Four-fold volume methanol precipitation was used to treat subunit influenza vaccine. The chromatographic conditions were as follows: column (Thermo BioBasic-4 C4 column 4.6 mm × 150 mm, 5 μm), mobile phase methanol: water 80:20 / v), flow rate: 1 ml / min, UV detector detection wavelength: 275 nm, column temperature: 33 ℃, sample volume: 100μl. The established HPLC method was validated. Results The established HPLC method showed good linearity in the concentration range of 5 ~ 2 500 μg / ml. The nonoxynol-9 was of good specificity. The detection of nonoxynol-9 in the inactivated The recoveries of influenza virus extracts and subunit influenza vaccines were about 100%. The intra- and inter-day precision of the non-phenyl alcohol ether-9 standard solution and the subunit influenza vaccine were all less than 3%. The detection limits The limit of quantification was 5μg / ml. Conclusion The established HPLC method is simple, rapid and accurate, and is suitable for the laboratory routine testing of lysozyme in subunit influenza vaccine.