Jiang Jia-Lu, Kang An, Li Qin, Song Meng-Meng, Han Hui-Fa, Cai Yu-Qing, Xu Zhui-Cheng, Cheng Hai-Bo, Sun Dong-Dong
School of Pharmacy, Nanjing University of Chinese Medicine Nanjing 210023, China.
Collaborative Innovation Center of Jiangsu Province of Cancer Prevention and Treatment of Chinese Medicine Nanjing 210023, China.
Zhongguo Zhong Yao Za Zhi. 2022 Jan;47(2):437-443. doi: 10.19540/j.cnki.cjcmm.20210901.201.
The present study developed an ultra-fast liquid chromatography coupled with triple quadrupole-linear ion trap composite mass spectrometry(UHPLC-QTRAP-MS) to simultaneously determine the content of potential active components in Scutellariae Barbatae Herba and also to provide a reference approach for screening out the differential quality control components among different batches of Scutellariae Barbatae Herba. Chromatographic separations were conducted on a Thermo Acclaim(TM) RSLC 120 C_(18) column(3.0 mm×100 mm, 2.2 μm) in a gradient program. The mobile phase consisted of 0.1% aqueous formic acid and acetonitrile, and the column temperature was maintained at 40 ℃. The flow rate was 0.4 mL·min(-1) and the injection volume was 2 μL. The targeted compounds were monitored in the multiple reaction monitoring(MRM) mode. The acquired data were processed by hierarchical cluster analysis(HCA) and partial least square discriminant analysis(PLS-DA). Sixteen compounds all showed good linear relationship within the corresponding linear ranges and the R~2 values were all higher than 0.993 2. The RSDs of precision, repeatability, and stability were less than or equal to 3.7%. Mean recovery rates were in the range of 95.67% and 104.8% with RSDs≤3.2%. According to HCA and PLS-DA, all samples were clustered into four categories. Scutellarin, acteoside, scutellarein, and scutebarbatine X(VIP>1) were considered as differential chemical markers in the four categories. In conclusion, the developed method can be used for the simulta-neous determination of the multiple components and quality control of Scutellariae Barbatae Herba.
本研究开发了一种超快速液相色谱-三重四极杆-线性离子阱复合质谱联用技术(UHPLC-QTRAP-MS),用于同时测定半枝莲中潜在活性成分的含量,并为筛选不同批次半枝莲中的差异质量控制成分提供参考方法。采用梯度洗脱程序,在Thermo Acclaim(TM) RSLC 120 C_(18)柱(3.0 mm×100 mm,2.2 μm)上进行色谱分离。流动相由0.1%甲酸水溶液和乙腈组成,柱温保持在40℃。流速为0.4 mL·min(-1),进样量为2 μL。目标化合物采用多反应监测(MRM)模式进行监测。采集的数据通过层次聚类分析(HCA)和偏最小二乘判别分析(PLS-DA)进行处理。16种化合物在相应线性范围内均呈现良好的线性关系,R~2值均高于0.993 2。精密度、重复性和稳定性的相对标准偏差(RSD)均小于或等于3.7%。平均回收率在95.67%至104.8%之间,RSD≤3.2%。根据HCA和PLS-DA分析,所有样品聚为四类。野黄芩苷、毛蕊花糖苷、黄芩素和半枝莲碱X(VIP>1)被认为是这四类中的差异化学标志物。综上所述,所建立的方法可用于半枝莲多种成分的同时测定和质量控制。