超高效液相色谱指纹图谱结合化学模式识别评价白蒲黄胶囊质量

Quality Evaluation of Baipuhuang Capsules by UPLC Fingerprint Combined with Chemical Pattern Recognition

目的 建立评价白蒲黄胶囊质量的超高效液相色谱(UPLC)指纹图谱和化学模式识别方法。方法 色谱柱为Waters Acquity BEH C_(18)柱(150 mm×2.1 mm,1.7μm),流动相为乙腈-0.02 mol/L磷酸二氢钾溶液(梯度洗脱),流速为0.3 mL/min,柱温为35℃,切换检测波长210 nm及323 nm,进样量为2μL。建立2个厂家10批样品的指纹图谱,并进行相似度评价、聚类分析(CA)和正交偏最小二乘判别分析(OPLS-DA),筛选不同批次样品的差异性标志物。结果 方法学考察结果显示,精密度、重复性、稳定性试验结果的RSD均小于2.0%。10批样品共标定了19个共有峰;通过与对照品比对,指认7个成分,分别为盐酸黄柏碱(4号峰)、绿原酸(6号峰)、咖啡酸(10号峰)、菊苣酸(11号峰)、黄芩苷(13号峰)、盐酸小檗碱(17号峰)、白头翁皂苷B_(4)(19号峰)。10批样品的指纹图谱相似度均不低于0.978。10批样品按厂家聚为2类;在OPLS-DA模型下,以变量重要性投影值大于1筛选出4个差异性标志物,分别为白头翁皂苷B_(4)(19号峰)、5号峰、菊苣酸(11号峰)、绿原酸(6号峰)。结论 该方法操作简单、结果准确,可客观评价白蒲黄胶囊的质量。

Objective To establish an ultra-high performance liquid chromatography (UPLC) fingerprint and chemical pattern recognition method for evaluating the quality of Baipuhuang Capsules.MethodsThe chromatography column was Waters Acquity BEH C_(18)column (150 mm×2.1 mm,1.7μm),the mobile phase was acetonitrile -0.02 mol/L potassium dihydrogen phosphate solution (gradient elution),the flow rate was 0.3 mL/min,the column temperature was 35℃,the switching wavelengths were 210 nm and 323 nm,and the injection volume was 2μL.The fingerprints of ten batches of samples from two manufacturers were established,and the similarity evaluation,clustering analysis (CA),and orthogonal partial least squares-discriminant analysis(OPLS-DA) were conducted to screen for differential biomarkers in different batches of samples.ResultsThe methodological examination results showed that the RSDs of the precision,repeatability,and stability test results of the seven components were all lower than 2.0%.Nineteen common peaks were calibrated in ten batches of samples.By comparing with the reference substance,seven components were identified,namely phellodendrine hydrochloride (peak 4),chlorogenic acid (peak 6),caffeic acid (peak 10),chicoric acid (peak 11),baicalin (peak 13),berberine hydrochloride (peak 17),and pulchinenoside B_(4)(peak 19).The similarity of the fingerprints of ten batches of samples was 0.978 or above.Ten batches of samples were classified into two categories according to the manufacturers.Under the OPLS-DA model,four differential biomarkers were screened based on the variable importance in projection (VIP) value greater than 1,namely pulchinenoside B_(4)(peak 19),peak 5,chicoric acid (peak 11),and chlorogenic acid(peak 6).ConclusionThe method is simple and accurate,which can objectively evaluate the quality of Baipuhuang Capsules.