HPLC法测定兔眼房水中黄芩苷浓度的不确定度评定

Uncertainty Evaluation of the Determination of Baicalin in Rabbit Aqueous Humor by HPLC

目的:评定采用高效液相色谱(HPLC)法测定兔眼房水中黄芩苷浓度的不确定度。方法:对HPLC法测定兔眼房水中黄芩苷浓度的全过程进行分析,对于分析过程中称量、溶液配制、分析仪器、标准曲线拟合等影响因素引起的不确定度进行了评定,用A类评定程序评价了分析过程中随机效应引起的不确定度,用B类评定程序评价了分析过程中其他因素引起的不确定度,最后根据各分量计算出合成不确定度并进行了扩展。结果:置信概率P为95%时,兔眼房水中低、中、高浓度黄芩苷(0.04、0.125、0.25μg·mL-1)的扩展不确定度分别为0.003、0.01、0.02μg·mL-1。结论:本方法适用于HPLC法测定兔眼房水中黄芩苷浓度的不确定度评定,该方法的确立对于复杂生物基质分析过程的不确定度研究具有重要意义。

OBJECTIVE： To evaluate the uncertainty of the determination of baicalin in rabbit aqueous humor by HPLC. METHODS： The determination process of baicalin in rabbit aqueous humor by HPLC was analyzed. The uncertainty caused by vari- ous factors in the whole process of determination, including balancing, solution preparation, calibration fitting and the equipment, were estimated. The uncertainty caused by random effects was evaluated with type A evaluation process and others were by type B. The combined uncertainty was calculated with all the components. RESULTS： The expanded uncertainty for low, medium and high level of baicalin （0.04,0.125,0.25μg.mL^-1） were 0.003, 0.01 and 0.02μg.mL^-1, respectively（P：95%）. CONCLUSION： The method is applicable for the uncertainty evaluation of baicalin determination in rabbit aqueous humor by HPLC. It provides an im- portant reference for the uncertainty evaluation for the determination in complex bio-matrix.