人用布氏菌疫苗浓度测定分光光度法的建立

目的研究人用布氏菌疫苗浓度与吸光度（A）值之间的相关性,建立用分光光度法测定待测疫苗吸光度值,经参考曲线得出其浓度的方法。方法确定分光光度法测定布氏疫苗浓度的特异性、测定波长及线性范围。4个专业实验室经对7批成品疫苗进行比浊,测定与细菌浊度标准管（相当于布氏菌浓度25亿.mL-1）浊度一致菌液的吸光度值及标准管对应浓度±20%（即30亿.mL-1和20亿.mL-1）菌液的吸光度值,根据该结果制备布氏菌疫苗浓度测定用参考品。4个实验室应用该参考品分别对布氏菌疫苗进行协作测定,确定分光光度法测定布氏菌疫苗浓度的可行性。结果确定600 nm作为吸光度测定波长,布氏菌疫苗浓度在10~50亿.mL-1之间时吸光度值与菌液浓度之间有很好的相关性（r=0.999,自由度=3,P〈0.01）。成功制备了3种浓度（即20、25、30亿.mL-1）的布氏菌疫苗浓度测定用参考品,其吸光度值分别为0.384、0.484和0.575。经4个实验室应用该参考品对布氏菌疫苗进行协作测定,测定误差小于5%。结论分光光度法具有良好的重现性,适用于布氏菌疫苗的浓度测定。

Establishment of Spectrophotometric Method for Determining the Concentration of Brucellosis Vaccine

Objective To study the correlation between the concentration of brucellosis vaccine and absorbance value and to establish a method for determining the concentration of brucellosis vaccine by spectrophotometry.Methods Firstly,the specificity of spectrophotometry,measurement wavelength and linear range were determined.The absorbance values of vaccines with different concentrations（20×108·mL-1,25×108·mL-1,30×108·mL-1） were determined,based on which reference materials for measuring the concentration of brucellosis vaccine was established.Finally,reference materials were evaluated by measuring three batches of vaccine.Results Between 10×108·mL-1 and 50×108·mL-1,the absorbance value was highly correlated with the concentration of brucellosis vaccine at 600nm（r=0.999,df=3,P0.01）.Reference materials were successfully established,and absorbance values of which were 0.384,0.484 and 0.575.By measuring 3 batches of vaccine,the determination error was less than 5%.Conclusion Spectrophotometric method possesses excellent reproducibility and reliability to determine the concentration of brucellosis vaccine.