College of Medical Imaging Laboratory and Rehabilitation, Xiangnan University, Chenzhou 423000, Hunan Province, PR China; Zhuhai Hengqin HEC Cordyceps Co., Ltd, Zhuhai 519031, Guangdong Province, PR China.
Zhuhai Hengqin HEC Cordyceps Co., Ltd, Zhuhai 519031, Guangdong Province, PR China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Oct 15;1247:124322. doi: 10.1016/j.jchromb.2024.124322. Epub 2024 Sep 16.
Hypericum japonicum is a traditional folk medicine with various bioactivities such as hepatoprotective, antioxidant, and anti-tumorous. The antioxidant effect of H. japonicum is one of the most prominent effects due to its responsibility for many of its activities. To clarify active natural substance, the antioxidant properties of H. japonicum were preliminarily assessed by ferric reducing-antioxidant power (FRAP), 2,2-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) and Oxygen radical absorbance capacity (ORAC), as well as superoxide dismutase (SOD). Then, a straightforward and effective method named online liquid extraction-high performance liquid chromatography combined with ABTS antioxidant assay and mass spectrometry (OLE-HPLC-ABTS/Q-TOF-MS) was developed to swiftly and directly discover the antioxidants in H. japonicum. Using mobile phase as extraction and separation reagent, coupled with online activity analysis and compounds identification by high-resolution MS, the online system enables rapid screening of natural antioxidant bioactives from complex mixture. By using it, a total of 9 compounds including flavonoids and phenolic acids characterized by retention time, precise mass, and fragmentation ions in MS/MS spectra showed antioxidant action. Finally, the antioxidant and SOD activity of main found active compounds were validated by in vitro experiment assay and molecular docking. In summary, these results suggested that H. japonicum could be considered as a potential source of natural antioxidants, and the online integrated system might become a promising candidate for the natural antioxidants discovery in the future.
贯叶金丝桃是一种传统的民间药物，具有多种生物活性，如保肝、抗氧化和抗肿瘤。由于贯叶金丝桃对其许多活性负有责任，因此其抗氧化作用是最突出的作用之一。为了阐明活性天然物质，通过铁还原抗氧化能力（FRAP）、2,2-连氮-双-3-乙基苯并噻唑啉-6-磺酸（ABTS）和氧自由基吸收能力（ORAC）以及超氧化物歧化酶（SOD）初步评估了贯叶金丝桃的抗氧化特性。然后，开发了一种简单有效的方法，称为在线液提取-高效液相色谱结合 ABTS 抗氧化测定和质谱法（OLE-HPLC-ABTS/Q-TOF-MS），以快速直接地发现贯叶金丝桃中的抗氧化剂。该在线系统使用流动相作为提取和分离试剂，结合在线活性分析和通过高分辨率 MS 进行化合物鉴定，能够快速筛选复杂混合物中的天然抗氧化生物活性物质。通过使用该系统，总共发现了 9 种化合物，包括黄酮类化合物和酚酸类化合物，它们的特征在于保留时间、精确质量和 MS/MS 谱中的碎片离子，表现出抗氧化作用。最后，通过体外实验测定和分子对接验证了主要发现的活性化合物的抗氧化和 SOD 活性。综上所述，这些结果表明贯叶金丝桃可被视为天然抗氧化剂的潜在来源，而在线集成系统可能成为未来天然抗氧化剂发现的有前途的候选者。
