Key Laboratory of TCM Quality Control Technology, Shandong Analysis and Test Center, Qilu University Of Technology (Shandong Academy of Sciences), Jinan, 250014, China.
State Key Laboratory and Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
J Pharm Biomed Anal. 2019 Oct 25;175:112758. doi: 10.1016/j.jpba.2019.07.006. Epub 2019 Jul 6.
Flos Lonicerae Japonicae(Jinyinhua) possesses clearing heat and detoxification activity, and has been used as a traditional Chinese medicine to treat influenza for many years. Due to the complex chemical composition and diverse content of Jinyinhua, especially the many trace ingredients, the effective components are unknown. In this study, an improved two-dimensional high performance liquid chromatography-ultrafiltration combined with electrospray ionization-time-of-flight/mass spectroscopy (ESI-TOF/MS) approach was designed and used for the enrichment, screening and characterization of minor neuraminidase inhibitors in Jinyinhua. In the first dimension, semi-prep-HPLC was employed for the preliminary separation of different polarity components and enrichment of low content components from Jinyinhua extract. In the second dimension, hydrophilic interaction liquid chromatography and reverse phase-HPLC were used to separate the different polar fractions, respectively. The fractions then underwent ultrafiltration and ESI-TOF/MS for the comprehensive screening and characterization of potential neuraminidase inhibitors. As a result, a total of 44 compounds were found to have neuraminidase inhibitory activity, and 22 of these compounds were preliminarily identified by accurate molecular weight and UV absorption data compared with standards and references. The activity of 16 of these compounds was verified by the neuraminidase inhibition assay. This study provides support for the rapid screening of minor neuraminidase inhibitors from complex natural medicines.
金银花具有清热解毒的功效，多年来一直被用作治疗流感的中药。由于金银花的化学成分复杂，含量多样，尤其是许多痕量成分，其有效成分尚不清楚。本研究设计了一种改进的二维高效液相色谱-超滤结合电喷雾电离-飞行时间/质谱（ESI-TOF/MS）方法，用于从金银花中富集、筛选和表征微量神经氨酸酶抑制剂。在第一维中，半制备 HPLC 用于初步分离不同极性成分，并从金银花提取物中富集低含量成分。在第二维中，亲水作用液相色谱和反相-HPLC 分别用于分离不同极性的馏分。然后对馏分进行超滤和 ESI-TOF/MS 综合筛选和潜在神经氨酸酶抑制剂的表征。结果，共发现 44 种化合物具有神经氨酸酶抑制活性，其中 22 种化合物通过与标准品和参考文献相比准确的分子量和紫外吸收数据进行了初步鉴定。通过神经氨酸酶抑制试验验证了其中 16 种化合物的活性。本研究为从复杂天然药物中快速筛选微量神经氨酸酶抑制剂提供了支持。
