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目的研究疫苗中硫柳汞含量对劳里测定(Lowry Assay,Lowry)法测定蛋白质含量结果的影响,为准确测定新型甲型H1N1流行性感冒(甲流)疫苗蛋白质含量提供依据。方法配制终浓度分别为30μg/ml(微克/毫升)与60μg/ml的甲流病毒抗原、人血清白蛋白及重组乙型肝炎病毒表面抗原溶液(均含有终浓度分别为10、20、40、60、80、100μg/ml的硫柳汞),于不同时间点使用Lowry法(《中国药典》2010年版三部)测定各溶液的蛋白质含量,将结果进行比较。结果以Lowry法测定蛋白质含量,30μg蛋白/ml组回收率在125.85%～284.14%,硫柳汞含量与回收率呈正相关,相关系数(Coefficient of Correlation,r)为0.992～0.997;去除硫柳汞影响后,回收率99.25%～104.92%。60μg蛋白/ml组回收率112.12%～191.60%,硫柳汞含量与回收率呈正相关,r值为0.993～0.997;去除硫柳汞影响后,回收率97.35%～103.79%。结论硫柳汞可使Lowry法测定的蛋白质含量结果偏高,且硫柳汞含量与蛋白质含量偏高程度呈高度正相关,通过定量测定去除其影响后,可以准确测出甲流疫苗中的蛋白质含量,为甲流疫苗质量评价提供依据。
Objective To study the effect of thimerosal in vaccine on the determination of protein content by Lowry Assay (Lowry) method and provide the basis for accurate determination of the protein content of the new influenza A (H1N1) influenza vaccine. Methods The final concentration of 30μg / ml (μg / ml) and 60μg / ml of influenza A virus antigen, human serum albumin and recombinant hepatitis B virus surface antigen solution (containing final concentrations were 10,20,40, 60, 80, 100μg / ml thimerosal). The protein content of each solution was determined by Lowry method (“Chinese Pharmacopoeia 2010”) at different time points. The results were compared. Results The protein content was determined by Lowry method. The recoveries of thimerosal were in the range of 125.85% ~ 284.14% at the concentration of 30μg protein / ml, and the correlation coefficients (r) were 0.992 ~ 0.997. After removing the thimerosal effect, Rate of 99.25% ~ 104.92%. The recoveries of 60μg protein / ml were 112.12% ~ 191.60%. The contents of thimerosal were positively correlated with the recoveries, and the values of r were 0.993 ~ 0.997. The recoveries were 97.35% ~ 103.79% after thimerosal was removed. Conclusions Thimerosal can make the protein content determined by Lowry method high, and the content of thimerosal has a high positive correlation with the high protein content. After removing the influence by quantitative determination, the protein content in the A flu vaccine can be accurately determined. Provide the basis for the quality evaluation of the flow vaccine.