特征图谱结合化学模式识别筛选不同产地茯苓药材差异质量标志物

Screening of differential quality markers of Poria from different regions by characteristic fingerprints and chemical pattern recognition

目的:利用化学模式识别技术,对茯苓药材HPLC特征图谱数据进行分析,筛选差异质量标志物并建立定量分析策略,为茯苓质量标准研究提供科学依据。方法:采用Welch Ultimate Plus C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈(含0.2%四氢呋喃)-0.1%磷酸水溶液,梯度洗脱,检测波长222 nm,体积流量1 mL·min^(-1),柱温30℃,进样量10μL,建立茯苓药材特征图谱。运用相似度分析、主成分分析(principal component analysis,PCA)、聚类分析(cluster analysis,CA)和正交偏最小二乘-判别分析(orthogonal partial least squares discrimination analysis,OPLS-DA)等模式识别方法,筛选不同产地茯苓的差异质量标志物,并按照《中华人民共和国药典》中“分析方法验证指导原则”的要求,进行定量研究。结果:15批茯苓药材的相似度在0.935~0.998,共标定8个共有峰,指认出6个色谱峰;PCA结果显示样品按产地呈现聚类趋势,CA结果与PCA一致,通过OPLS-DA筛选茯苓酸作为差异质量标志物;建立的茯苓酸含量测定方法稳定、可靠且耐用性良好,符合药典要求。结论:通过特征图谱结合化学模式识别技术的分析策略,可快速有效地筛选不同产地茯苓药材的差异质量标志物,为茯苓药材质量标准的建立提供参考。

Objective:The chemical pattern recognition technology was used to analyze the HPLC characteristic fingerprints data of Poria,screen the differential quality markers and perform quantitative analysis,so as to provide scientific basis for quality evaluation of Poria.Methods:Welch Ultimate Plus C_(18)chromatographic column(250 mm×4.6 mm,5μm)was used.Acetonitrile(containing 0.2%tetrahydrofuran)-0.1%phosphoric acid aqueous solution with gradient elution was used as mobile phase.The fingerprint of Poria was established with detection wavelength set at 222 nm,flow rate at 1 mL·min^(-1),column temperature at 30℃,injection volume at 10μL.The differential quality markers were identified by similarity analysis,principal component analysis(PCA),cluster analysis(CA)and orthogonal partial least squares-discriminant analysis(OPLS-DA).The quantitative research was carried out in accordance with the requirements of“Guidelines for Validation of Analytical Methods”in Chinese Pharmacopoeia.Results:The similarity of 15 batches of Poria was between 0.935~0.998.Eight common peaks were calibrated and 6 were identified;The samples were classified according to the origins from PAC results,which is consistent with CA result.Pachymic acid was screened as the differential quality marker of Poria by OPLS-DA.The established method for the determination of pachymic acid was stable,reliable,durable,which meets the requirements of the Pharmacopoeia.Conclusion:The methods of characteristic fingerprint combined with chemical pattern recognition technology can effectively screen the differential quality marker of Poria from different regions,providing a reference for the establishment of quality standards for Poria.