女贞子-墨旱莲药对提取物的指纹图谱和化学成分鉴定研究  

Fingerprint and Identification of Chemical Constituents of the Extract of Fructus Ligustri Lucidi-Ecliptae Herba

作　　者：高雨桐 宋秀苹 李楠[1] 尚涌琳 韩飞[1] GAO Yutong;SONG Xiuping;LI Nan;SHANG Yonglin;HAN Fei(School of Pharmacy,Shenyang Pharmaceutical University,Shenyang 110016,China)

机构地区：[1]沈阳药科大学药学院,辽宁沈阳110016

出　　处：《南京中医药大学学报》2024年第4期399-412,共14页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：辽宁省科技厅面上项目(2021-MS-220);辽宁省教育厅(JYTMS20231378)。

摘　　要：目的建立女贞子-墨旱莲药对提取物的HPLC指纹图谱以控制其质量,并对提取物中所含有的化学成分进行结构鉴定。方法采用HPLC法,固定相为ZORBAX Extend C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-体积分数为0.1%甲酸溶液梯度洗脱,流速为1.0 mL·min^(-1),检测波长为265 nm,柱温为35℃,进样量为10μL;建立15批不同产地女贞子-墨旱莲药对提取物的HPLC指纹图谱,对其进行相似度评价、聚类分析和正交偏最小二乘方判别分析;采用超高效液相色谱-静电场轨道阱高分辨质谱(UPLC-Q-Exactive Orbitrap MS)联用技术,分别在正、负离子扫描模式下对提取中的化学成分进行检测,基于高分辨质谱一级和二级数据进行结构鉴定。结果由不同产地药材随机组合的15批女贞子-墨旱莲药对提取物的相似度介于0.865~0.992之间,药对指纹图谱中共确定了16个共有峰,指认了其中7个色谱峰;液质联用共检测到55个化合物,采用对照品准确鉴定了7个成分的化学结构,并推测了其它48个化合物的可能化学结构。结论建立的指纹图谱方法简便可靠,重复性好,可用于女贞子-墨旱莲药对提取物的质量控制研究;UPLC-Q-Exactive Orbitrap MS联用技术可实现对女贞子-墨旱莲药对提取物中化合物的结构鉴定,为其后续研究奠定了基础。OBJECTIVE To establish high performance liquid chromatography(HPLC)fingerprints of the extract of Fructus Ligustri Lucidi-Ecliptae Herba and to identify the structure of its chemical constituents.METHODS The chromatographic separation was performed on ZORBAX Extend C_(18)(250 mm×4.6 mm,5μm)column with the mobile phase consisting of acetonitrile and 0.1%formic acid solution in gradient elution mode at a flow rate of 1.0 mL·min^(-1).The optimum column temperature was set at 35℃,the detection wavelength was 265 nm,and the injection volume was 10μL.The HPLC fingerprints of 15 batches of herb pair from different origins were established.Then,similarity evaluation,cluster analysis,and orthogonal partial least squares discriminant analysis were carried out.The chemical structures of the constituents of the herb pair were identified based on the high-resolution mass spectrometry utilizing the UPLC-Q-Exactive Orbitrap MS in positive and negative-ion modes.RESULTS As a result,the similarity of the 15 batches of samples varied from 0.865 to 0.992.A total of 16 common peaks were identified in the fingerprints,and 7 of them were identified by comparision with the reference substances.Furthermore,55 chemical compounds were detected,and 7 of them were accurately identified based on the reference substances,and the chemical structures of the rest 48 components were temporarily speculated.CONCLUSION The fingerprint method established in this study is simple,reliable,and reproducible,and can be further used for the quality control of Fructus Ligustri Lucidi-Ecliptae Herba herb pair.The developed UPLC-Q-Exactive Orbitrap MS method can be applied to identify the structures of chemical constituents in this herb pair and lay a foundation for further research.

关 键 词：墨旱莲 女贞子 指纹图谱 超高效液相 静电场轨道阱高分辨质谱 化学成分 

分 类 号：R284[医药卫生—中药学]