穿山甲色谱指纹谱的建立及其在基源鉴别和质量等级区分方面的应用  

Establishment of chromatographic fingerprint of Squama Manis and its applications in animal source identification and quality grade discrimination

作　　者：乔亚丽 刘喆 沈爱金[1] 郭志谋[1] 刘艳芳[1] 陈相银[3] 徐青[1] 梁鑫淼[1] QIAO Yali;LIU Zhe;SHEN Aijin;GUO Zhimou;LIU Yanfang;CHEN Xiangyin;XU Qing;LIANG Xinmiao(Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China;University of Chinese Academy of Sciences, Beijing 100049, China;Beijing Sanhe Pharmaceutical Co., Ltd., Beijing 100049, China)

机构地区：[1]中国科学院大连化学物理研究所,中国科学院分离分析化学重点实验室,辽宁大连116023 [2]中国科学院大学,北京100049 [3]北京三和药业有限公司,北京100049

出　　处：《色谱》2020年第12期1440-1448,共9页Chinese Journal of Chromatography

摘　　要：中药穿山甲为鳞鲤科动物穿山甲的鳞甲,常用于治疗乳汁不通、痈肿疮毒、风湿痹痛等症,饮片来源上经常存在基源不清、等级不明的问题。针对以上问题,目前已发展出了多种分析方法,包括薄层色谱、质谱和DNA检测,但由于特异性低、设备成本高、步骤繁琐等不足,以上方法均不适合作为穿山甲质量监测的常规手段,而色谱指纹谱技术恰好可以弥补以上不足。该文旨在建立穿山甲的色谱指纹谱,并探讨其在等级及基源鉴别方面的可行性。穿山甲粉末经1 mol/L HCl加热水解得供试液,色谱柱采用Waters Symmetry 300 C18,以0.1%(v/v)三氟乙酸/水、0.1%(v/v)三氟乙酸/乙腈为流动相,280 nm为检测波长,采用梯度洗脱获得穿山甲指纹谱。经考察,该方法的精密度、日内日间重复性及样品稳定性均表现良好(RSD<5%)。以12批中华穿山甲一等品的平均指纹谱为对照指纹谱,同时确认17个共有峰,继而以共有峰绝对峰面积为原始数据,计算样品指纹谱与对照指纹谱的相似度。结果表明,中华穿山甲与其他动物来源穿山甲的相似度不高于0.776,体现了相似度评价在基源鉴别上的有效性;但不同等级穿山甲之间交叉严重,表明相似度评价在等级鉴别上的局限性。以24批不同等级中华穿山甲为分析对象建立判别分析模型,经十折交叉验证,模型无偏差正确率为95.83%,说明该模型在等级区分上可行性很高。该文通过建立穿山甲的色谱指纹谱,并结合不同数据处理方法验证了其在基源鉴别和等级区分方面的可行性,为保证穿山甲临床用药的科学性、准确性提供新的思路。Squama Manis,or“Chuanshanjia”in Chinese,is a traditional Chinese medicine(TCM)for promoting blood circulation and reducing swelling and discharge;the only animal source used in TCM is the scales of Manis pentadactyla.However,in today’s pharmaceutical market,there are many scales from other species of the same genus that are difficult to distinguish from Squama Manis.High-quality and low-quality scales are also severely confused.To solve the above problems,various analytical methods have been developed,such as thin-layer chromatography,mass spectrometry and DNA detection.Owing to their low resolving ability,high equipment cost,and inconvenient operation,none of these methods are appropriate for routine identification of Squama Manis.A chromatographic fingerprint can comprehensively reflect the synergic action of multiple chemical compositions in TCM and has been widely used for the quality control of TCM.In the present study,we established a fingerprint of Squama Manis and explored its feasibility in identifying the origin and quality grade of scales.First,Squama Manis powder was hydrolyzed by hydrochloric acid(1 mol/L).Next,the extract was analyzed on a Symmetry 300 C18 column by linear gradient elution,using 0.1%trifluoroacetic acid(v/v)in water and 0.1%trifluoroacetic acid(v/v)in acetonitrile as the mobile phase and 280 nm as the detection wavelength.The established method was systematically validated,demonstrating good precision,repeatability and sample stability(relative standard deviation(RSD)<5%).Subsequently,samples of different sources and quality grades were distinguished by similarity evaluation and discrimination analysis based on the fingerprint data.In the similarity evaluation,the reference fingerprint was defined as the average fingerprint of twelve first-class samples,and seventeen chromatographic peaks were identified as common peaks.Similarities between the reference fingerprint and fingerprints with different base sources and quality grades were calculated using the absolute area of commo

关 键 词：中药 穿山甲 色谱指纹谱 判别分析 相似度评价 质量等级 基源 

分 类 号：O658[理学—分析化学]