大黄[庶虫]虫丸UPLC指纹图谱及化学模式识别研究

UPLC fingerprint analysis and chemical pattern recognition of Dahuang Zhechong Pill

目的建立大黄[庶虫]虫丸超高效液相色谱(UPLC)指纹图谱,辅以化学模式识别方法,为建立相应的质量评价体系提供思路和参考。方法色谱柱为Waters Acquity UPLC HSS T3柱(2.1 mm×100 mm,1.8μm),以乙腈(A)-0.2%甲酸(B)为流动相,梯度洗脱,根据各味药材主成分出峰顺序优化检测波长切换程序,柱温:35℃,流速:0.3 mL/min,应用"中药色谱指纹图谱相似度评价系统"(2012.1版)建立大黄[庶虫]虫丸对照指纹图谱并进行相似度评价,再运用ChemPattern化学计量学软件(2017年版)进行聚类分析、主成分分析(PCA)、偏最小二乘回归分析(PLS)和模式识别。结果共标定30个共有峰,根据色谱峰行为、超高效液相色谱-电喷雾离子源-四极杆飞行时间质谱联用技术(UPLC-ESI-QTOF-MS)对照品离子碎片及文献信息确认13个已知成分,并进行各峰归属。B、C、F、G厂家样品相似度均>0.90,PLS分析能较好地区分各个厂家样品;筛选出黄芩、大黄的多个主要成分及苦杏仁苷是强特征峰,聚类分析、PCA、PLS分析结果相互验证。结论大黄[庶虫]虫丸UPLC指纹图谱具有较好的稳定性和重复性,结合化学模式识别方法,能较为快速、准确地反映大黄[庶虫]虫丸的整体质量,可为其质量控制和评价提供依据。

Objective To establish UPLC fingerprint of Dahuang Zhechong Pill,as well as pattern recognition,and provide ideas and references for the establishment of corresponding quality evaluation system.Methods Waters Acquity UPLC HSS T3 column(2.1 mm×100 mm,1.8 μm)was taken,and acetonitrile-0.2% formic acid solution with gradient elution was used as mobile phase.The detection wavelength switching procedure was optimized according to the peak sequence of the main components of various medicinal materials.The column temperature was 35 ℃,and the flow rate was 0.3 mL/min.The TCM chromatographic fingerprint similarity evaluation system(version 2012.1)was used to establish the control fingerprint of Dahuang Zhechong Pill and evaluate the similarity,and then ChemPattern chemometrics software(version 2017)was used for cluster analysis,principal component analysis(PCA),partial least squares regression analysis(PLS)and pattern recognition.Results In the fingerprint,30 common peaks were confirmed.According to the chromatographic peak behavior,reference ion fragments in UPLC-ESI-QTOF-MS and literature information,13 known components were identified and other common peaks were attributed.The similarity of samples from B,C,F and G manufacturers was more than 0.90.PLS analysis could clearly distinguish the differences of samples from different manufacturers.Amygdalin and several main components of Scutellariae Radix and Rhei Radix et Rhizoma were selected as strong characteristic peaks.The results of cluster analysis,PCA and PLS were mutually verified.Conclusion The UPLC fingerprint of Dahuang Zhechong Pill shows good stability and repeatability.Combined with chemical pattern recognition method,it can quickly and accurately reflect the overall quality of Dahuang Zhechong Pill,which can provide the basis for its quality control and evaluation.