3种磷含量测定方法的比较

Comparison of three methods for determination of phosphorus content

目的比较3种A群脑膜炎球菌多糖蛋白结合物中磷含量测定方法的差异,为今后方法的选择及进一步优化提供参考。方法对《中国药典》三部（2010版）附录中的磷测定法（方法1）、高分子结合物含量测定法（方法 2）、文献中的磷测定法（方法 3）这3种方法的标准曲线、测定磷酸二氢钾对照品中磷含量、测定A群多糖、A群衍生物、A群结合物中磷含量以及标准曲线增加两个低浓度点的差异进行比较。结果不同人员采用同一方法绘制的标准曲线的斜率（b）的变异系数（CV）均小于10%,相关系数（r2）均在0.995以上,CV值在0-0.23%之间,其中方法 2标准曲线b值的CV值小于1%,且r2值与其他两种方法相比更稳定;3种方法测定同一份磷酸二氢钾对照品的结果与理论值相比,相对误差均在10%以下,其中方法 2的相对误差最小,为5.73%;3种方法测定A群多糖、A群衍生物、A群结合物中磷含量的CV值均小于10%;3种方法的标准曲线增加两个低浓度点后,b值及r2值的CV值均小于10%,与原标准曲线近似。结论 3种方法测定磷含量的标准曲线均具有良好的线性,不同操作人员的测定结果无明显差异,其中方法2测定A群脑膜炎球菌多糖、A群脑膜炎球菌多糖衍生物及A群脑膜炎球菌多糖蛋白结合物中的磷含量时,稳定性较好,操作简便易行。

Objective To compare the difference of three methods for determination of phosphorus content in group A meningococcal polysaccharide protein conjugate,and provide a basis for selection and optimization of determination method. Methods Phosphorus assay in Chinese Pharmacopoeia（Volume Ⅲ,2010 edition）（method 1）,high molecular conjugate content assay（method 2） and phosphorus assay in document（method 3） were compared in differences in standard curve,determination of phosphorus content in potassium dihydrogen phosphate control,group A meningococcal polysaccharide,group A meningococcal derivative and group A meningococcal polysaccharide protein conjugate as well as standard curve after addition of two low concentration points. Results The coefficients of variation（CVs） of slopes of standard curves plotted by various personnel by the same method were less than 10%. The coefficients of correlation（r2）were more than 0. 995,with CVs of 0 - 0. 23%. The CVs of b of standard curve of method 2 was less than 1%,while the r2 value was more stable than those of other two methods. The relative errors of determination results of the same potassium dihydrogen phosphate control by three methods to the theoretical values were less than 10%,of which that by method 2 was the lowest（5. 73%）. All the CVs of determination results of phosphorus contents in group A meningococcal polysaccharide,group A meningococcal derivative and group A meningococcal polysaccharide protein conjugate were less than 10%. After addition of two low concentration points,both the b and r2 of standard curves of the three methods were less than 10%,which were similar to those of original standard curves. Conclusion All the standard curves for determination of phosphorus content by three methods showed good linearity,and the determination results by different personnel showed no significant difference. However,method 2 was stable for determination of phosphorus contents in group A meningococcal polysaccharide,group A meningococcal derivative and group A meningococcal polysaccharide protein conjugate,which was easy to handle.