School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing 100029, China.
Beijing Research Institution of Chinese Medicine, Beijing University of Chinese Medicine, Beijing 100029, China.
Molecules. 2017 Oct 19;22(10):1756. doi: 10.3390/molecules22101756.
Menthae Haplocalycis herba, one kind of Chinese edible herbs, has been widely utilized for the clinical use in China for thousands of years. Over the last decades, studies on chemical constituents of Menthae Haplocalycis herba have been widely performed. However, less attention has been paid to non-volatile components which are also responsible for its medical efficacy than the volatile constituents. Therefore, a rapid and sensitive method was developed for the comprehensive identification of the non-volatile constituents in Menthae Haplocalycis herba using ultra-high performance liquid chromatography coupled with linear ion trap-Orbitrap mass spectrometry (UHPLC-LTQ-Orbitrap). Separation was performed with Acquity UPLC BEH C column (2.1 mm × 100 mm, 1.7 μm) with 0.2% formic acid aqueous solution and acetonitrile as the mobile phase under gradient conditions. Based on the accurate mass measurement (<5 ppm), MS/MS fragmentation patterns and different chromatographic behaviors, a total of 64 compounds were unambiguously or tentatively characterized, including 30 flavonoids, 20 phenolic acids, 12 terpenoids and two phenylpropanoids. Finally, target isolation of three compounds named Acacetin, Rosmarinic acid and Clemastanin A (first isolated from Menthae Haplocalycis herba) were performed based on the obtained results, which further confirmed the deduction of fragmentation patterns and identified the compounds profile in Menthae Haplocalycis herba. Our research firstly systematically elucidated the non-volatile components of Menthae Haplocalycis herba, which laid the foundation for further pharmacological and metabolic studies. Meanwhile, our established method was useful and efficient to screen and identify targeted constituents from traditional Chinese medicine extracts.
唇形科薄荷属植物薄荷，作为一种中国药食同源植物，在中国被广泛应用于临床已有数千年的历史。在过去的几十年中，对薄荷的化学成分的研究已经广泛开展。然而，与挥发性成分相比，对其非挥发性成分的研究却相对较少，而这些非挥发性成分也是其药效的重要组成部分。因此，本文建立了一种快速、灵敏的超高效液相色谱-线性离子阱-Orbitrap 质谱联用（UHPLC-LTQ-Orbitrap）法，用于薄荷非挥发性成分的综合鉴定。采用 Acquity UPLC BEH C 柱（2.1mm×100mm，1.7μm），以 0.2%甲酸水溶液和乙腈为流动相，在梯度条件下进行分离。基于精确质量测量（<5ppm）、MS/MS 碎片模式和不同的色谱行为，共鉴定出 64 种化合物，包括 30 种黄酮类化合物、20 种酚酸类化合物、12 种萜类化合物和 2 种苯丙素类化合物。最后，根据所得结果对三种化合物（即 Acacetin、迷迭香酸和 Clemastanin A）进行了目标分离，其中后两种化合物（Clemastanin A）为首次从薄荷属植物中分离得到，进一步证实了碎片模式的推断，并鉴定了薄荷中的化合物特征。本研究首次系统地阐明了薄荷的非挥发性成分，为进一步的药理学和代谢研究奠定了基础。同时，所建立的方法对于从中药提取物中筛选和鉴定靶向成分也是有效和高效的。
