分光光度法测定鼠疫疫苗浓度方法的建立

目的研究鼠疫疫苗浓度与吸光度（A）值之间的相关性，建立通过测定A值来计算鼠疫疫苗浓度的方法。方法确定分光光度法测定鼠疫疫苗浓度的特异性、测定波长及线性范围。4个专业的实验室经对9批成品疫苗进行比浊，测定与细菌浊度标准管（相当于鼠疫浓度7．0亿／ml）浊度一致菌液的A值及标准管对应浓度±20％（即5．6亿／ml和8.4亿／ml）菌液的A值，根据该结果制备鼠疫疫苗浓度测定用参考品。4个实验室应用该参考品对鼠疫疫苗进行协作测定，确定分光光度法测定鼠疫疫苗浓度的可行性。结果确定600nm作为吸光度测定波长，鼠疫疫苗浓度在2．6—10．5亿／ml之间时A值与菌液浓度之间有很好的相关性（r=0．9998，df=3，P〈0．01）。成功制备了3种浓度（即5．6、7．0和8．4亿/ml）的鼠疫疫苗浓度测定用参考品，其A值分别为0．444、0．557和0．651。经4个实验室应用该参考品对鼠疫疫苗进行协作测定，测定误差小于5％。结论分光光度法具有良好的重现性，适用于鼠疫疫苗浓度的测定。

Establishment of spectrophotometric method for determining the concentration of plague vaccine

Objective To study the correlation between the concentration of plague vaccine and Absorbance value and to establish a method for determining the concentration of plague vaccine. Methods Firstly, the specificity of spectrophotometry, measurement wavelength and linear range were determined. Secondly, four professional laboratories tested the concentrations of 9 batches of vaccine. The Absorbance values of vaccines with different concentrations (5.6 × 10⁸/ml, 7.0 × 10⁸/ml &; 8.4 × 10⁸/ml) were determined, based on which reference materials for measuring the concentration of plague vaccine was established. Finally, reference materials were evaluated by measuring three batches of vaccine. Results Between 2.6 × 10⁸/ml and 10.5 × 10⁸/ml, the Absorbance value was highly correlated with the concentration of plague vaccine at 600nm (r = 0.9998, df = 3, P < 0.01). Reference materials were successfully established, Absorbance value of which were 0.444, 0.557 and 0.651. 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 plague vaccine.