Quick identification of xanthine oxidase inhibitor and antioxidant from Erycibe obtusifolia by a drug discovery platform composed of multiple mass spectrometric platforms and thin-layer chromatography bioautography
Quick identification of xanthine oxidase inhibitor and antioxidant from Erycibe obtusifolia by a drug discovery platform composed of multiple mass spectrometric platforms and thin-layer chromatography bioautography
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DOI：
10.1002/jssc.201400342
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发表时间：
2014-08-01
影响因子：
3.1
通讯作者：
Wang, Zhengtao
中科院分区：
文献类型：
--
作者：
Chen, Zhiyong;Tao, Hongxun;Wang, Zhengtao
作为嘌呤代谢过程的最后一步，黄嘌呤氧化酶催化次黄嘌呤和黄嘌呤氧化成尿酸。我们的研究已经证明沙枣具有黄嘌呤氧化酶抑制活性。本论文的目的是描述一种基于多个质谱平台和薄层色谱生物自显影相结合的新策略，以有效地筛选黄嘌呤氧化酶的抑制和抗氧化特性。这一战略是通过以下步骤实现的。(I)用液相色谱-质谱法控制的自动纯化系统将蛇床子叶提取物分离成多个组分。(2)用薄层层析生物自显影法测定鬼针草有效部位。(3)利用实时光谱分析中直接分析提供的信息确定主要活性化合物的结构。(4)用高效液相色谱法计算各化合物对黄嘌呤氧化酶的IC50值。以淫羊藿茎为实验材料，筛选出7个目标峰，分别为黄嘌呤氧化酶抑制剂或抗氧化剂。我们的筛选策略允许对小分子进行快速分析，几乎不需要样品准备，并且可以在一周内完成，使其成为识别不稳定化合物的有用分析方法，并为夜来香作为痛风和氧化应激相关疾病的天然药物提供经验基础。
As a final step of the purine metabolism process, xanthine oxidase catalyzes the oxidation of hypoxanthine and xanthine into uric acid. Our research has demonstrated that Erycibe obtusifolia has xanthine oxidase inhibitory properties. The purpose of this paper is to describe a new strategy based on a combination of multiple mass spectrometric platforms and thin-layer chromatography bioautography for effectively screening the xanthine oxidase inhibitory and antioxidant properties of E. obtusifolia. This strategy was accomplished through the following steps. (i) Separate the extract of E. obtusifolia into fractions by an autopurification system controlled by liquid chromatography with mass spectrometry. (ii) Determine the active fractions of E. obtusifolia by thin-layer chromatography bioautography. (iii) Identify the structure of the main active compounds with the information provided by direct analysis in real timemass spectrometry. (iv) Calculate the IC50 value of each compound against xanthine oxidase using high-performance liquid chromatography. Using the caulis of E. obtusifolia as the experimental material, seven target peaks were screened out as xanthine oxidase inhibitors or antioxidants. Our screening strategy allows for rapid analysis of small molecules with almost no sample preparation and can be completed within a week, making it a useful assay to identify unstable compounds and provide the empirical foundation for E. obtusifolia as a natural remedy for gout and oxidative-stress-related diseases.