清开灵注射液中间体银黄液中黄芩苷含量近红外测定方法的建立和验证

Development and validation of a NIR quantification method for the determination of baicalin in intermediates of Qingkailing Injection

目的:建立清开灵注射液生产过程中间体银黄液中黄芩苷的近红外定量模型,并对其进行方法学验证。方法:采用近红外透射模式,以高效液相色谱分析值为参考值,采用偏最小二乘回归法(PLS)建立黄芩苷含量的近红外定量校正模型。采用相关系数(R)、校正标准差(SEC)、预测标准差(SEP)等参数评价模型,并运用AP方法验证模型。结果:所建模型的Rcal和Rval为0.9978和0.9995,SEC和SEP为723.0μg/mL和513.8μg/mL,模型预测效果良好;在β-期望容差区间为90%,容许极限为15%,风险性为10%的约束下,该模型各项分析指标满足分析要求,最低定量限为8 025μg/mL。结论:当黄芩苷含量大于8 025μg/mL时,所建近红外分析模型稳健可靠,可用于清开灵注射液生产过程中间体银黄液中黄芩苷快速准确定量。

Objective： The feasibility of near-infrared spectroscopy（NIRS） for baicalin content analysis in intermediates of Qingkailing Injection was verified in this work.Methods： The NIR spectra were collected in the transmission mode.The reference method used for the baicalin determination was HPLC assay recommended by the Chinese Pharmacopoeia（CHP,2010 Edition） for Qingkailing Injection.The model was developed based on partial least squares regression（PLS） and the R,standard error of calibration and prediction（SEC and SEP） was used for the evaluation of the model adequacy.A novel approach based on accuracy profile which allowed the acquisition of the lower limit of quantification（LLOQ） was used to validate the robustness and accuracy of PLS model.Results： Rcal,Rval,SEC and SEP were 0.9978,0.9995,723.0 μg/mL and 513.8 μg/mL.The acceptance limits were set at ±15% while the maximum risk to obtain results outside these acceptance limits was set at 10%.The accuracy profile showed that PLS model was able to determine baicalin content,whose LLOQ was about 8 025 μg/mL.Conclusion： The conclusion illuminated the feasibility of the NIR method when the content of baicalin is greater than 8 025 μg/mL,and provided a guideline of NIR accurate measurement in rapid analysis of intermediates of Qingkailing Injection.