UHPLC-MS/MS同时测定金线兰中7种成分含量及其分布规律研究  

Simultaneous determination of seven components and their distribution in Anoectochilus roxburghii by UHPLC-MS/MS

作　　者：吕欣锴 刁海欣 周丽思[1] 郭顺星[1] Lü Xin-kai;DIAO Hai-xin;ZHOU Li-si;GUO Shun-xing(Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100193,China)

机构地区：[1]中国医学科学院、北京协和医学院药用植物研究所,北京100193

出　　处：《药学学报》2023年第1期201-207,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81973426,81903749);中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-031)。

摘　　要：基于液质联用(UHPLC-MS/MS)技术,建立快速同时定量金线兰[Anoectochilus roxburghii(Wall.)Lindl.]中7种不同极性成分(金线莲苷、水仙苷、槲皮素、芦丁、异鼠李素-3-O-葡萄糖苷、槲皮素-7-O-葡萄糖苷、山柰酚-3-O-芸香糖苷)的方法。采用Waters Acquity UPLC BEH Shield RP18(2.1 mm×100 mm;1.7μm)色谱柱,以乙腈为流动相A,0.1%甲酸-水溶液为流动相B,0.2 mL·min^(-1)的流速进行梯度洗脱,柱温为30℃,进样量为2μL,分析时间为8 min。质谱以电喷雾电离源(electron spray ionization,ESI),正、负离子模式同时检测,优化各成分最佳质谱条件。结果发现,7种成分在测定浓度范围内线性关系良好,相关系数均≥0.9980,稳定性、精密度和重复性良好,平均加样回收率为97.71%~103.33%。经聚类热图和RDA(redundancy analysis)分析,发现金线莲苷主要分布在茎中,叶中次之,根中最少。生长6个月的金线兰茎中金线莲苷含量增长显著,而根与叶中增长较少。黄酮及黄酮苷类成分主要分布在叶。本文所建立的液质联用方法准确且灵敏,稳定性与重复性好,可用于金线兰的质量控制,并为建立更全面的金线莲药材质量检测方法提供参考。Based on ultra high-performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS),a rapid and simultaneous quantitative method for the measurement of seven components(kinsenoside;rutin;kaempferol-3-O-rutinoside;quercimeritrin;narcissin;isorhamnetin-3-O-glucoside;quercetin)of A.roxburghii was established.The separation was performed over 8.0 minutes on a Waters Acquity UPLC BEH Shield RP18(2.1 mm×100 mm;1.7μm)with a mobile phase consisting of acetonitrile(A)and 0.1%formic acid water solution(B)with gradient elution at a flow rate of 0.2 mL·min^(-1);the column temperature was 30℃and the injection volume was 2μL.Electrospray spray ionization source(ESI source)was used for mass spectrometry,and positive and negative ion modes were detected at the same time.The results showed good linearity(R2≥0.9980),with good precision,repeatability and stability,and the average recovery was 97.71%-103.33%.Through cluster heat map and redundancy analysis,we found that kinsenoside was mainly distributed in stems,followed by leaves,and the lowest content in roots.The content of kinsenoside increased significantly in the stems of plants 6 months,but less change was evident in the roots and leaves.Flavonoids and flavonoid glycosides were mainly distributed in leaves.The UHPLC-MS/MS method established in this paper can be used for the quality control of A.roxburghii and provides a reference for establishing a more comprehensive quality detection method for this medicinal.

关 键 词：金线兰 开唇兰属 金线莲苷 UHPLC-MS/MS 含量测定 分布规律 

分 类 号：R917[医药卫生—药物分析学]