用大孔树脂联合HSCCC纯化蒲公英草药片中的菊苣酸及其对人角质形成细胞的抗氧化能力

IF 2.8 3区 医学 Q2 BIOCHEMICAL RESEARCH METHODS
Ziwei Liang, Mengqi Wu, Jingying Xu, Yuxuan Liu, Qifeng Han, Xiang Yang, Wei Xia, Wenqing Zhang
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引用次数: 0

摘要

蒲公英(Taraxacum mongolicum)——黄芪(Mazz.)是一种传统的中草药,富含黄酮类和多酚等有效成分,用于治疗肿胀和炎症相关疾病。本研究采用HPD-500树脂柱层析法对蒲公英草药片提取物中的总黄酮和多酚进行了纯化,纯化后提取物E80%的总黄酮纯度为纯化前的5.3倍,达到43.98%。用UHPLC-QTOF-MS对提取液E80%进行分析,鉴定出咖啡酸、绿原酸、菊苣酸等8种酚酸类化合物。建立了高速逆流色谱分离E80%中菊酯酸的方法。选择正己烷-乙酸乙酯-甲醇- 0.5%醋酸水溶液(2:7:2:7,v/v/v/v)作为溶剂体系。在流速为2.0 mL/min,转速为850 r/min,系统温度为35℃,上样量为100 mg的条件下,得到纯度为95.44%的Chicoric酸。体外生物实验表明,菊酸对sds诱导的人角质形成细胞具有显著的抗氧化活性,可促进细胞增殖,提高超氧化物歧化酶(SOD)活性,降低活性氧(ROS)水平,稳定线粒体膜电位,抑制细胞凋亡。这些结果为天然抗氧化化合物的工业分离奠定了理论基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Purification of chicoric acid from dandelion herbal pieces using macroporous resin combined with HSCCC and its antioxidant capacity in human keratinocytes

Purification of chicoric acid from dandelion herbal pieces using macroporous resin combined with HSCCC and its antioxidant capacity in human keratinocytes
Dandelion (Taraxacum mongolicum Hand. - Mazz.) is a traditional Chinese medicinal herb rich in active ingredients such as flavonoids and polyphenols, which are used to treat swelling and inflammation-related diseases. In the present study, total flavonoids and polyphenols in dandelion herbal pieces (Taraxaci Herba) extracts were purified by HPD-500 resin column chromatography, and the total flavonoid purity in purified extracts E80% was 5.3 times that of before purification, reaching 43.98 %. The purified extracts E80% were analyzed by UHPLC–QTOF–MS and 8 phenolic acid compounds were identified, including caffeic acid, chlorogenic acid, and chicoric acid. Moreover, high-speed countercurrent chromatography method was established to separate chicoric acid from E80%. n-hexane-ethyl acetate-methanol-0.5 % acetic acid in water (2:7:2:7, v/v/v/v) was selected as the solvent system. Chicoric acid with a purity of 95.44 % was obtained with the flow rate of 2.0 mL/min, the speed of 850 r/min, the system temperature of 35 °C and sample loading of 100 mg. In vitro bioassay showed that chicoric acid presented impressive antioxidant activities in SDS-induced human keratinocytes by promoting cell proliferation, increasing superoxide dismutase (SOD) activity, reducing the reactive oxygen species (ROS) level, stabilizing the mitochondrial membrane potential, and inhibiting cell apoptosis. Collectively, these results lay a theoretical basis for the industrial separation of natural antioxidant compounds.
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来源期刊
Journal of Chromatography B
Journal of Chromatography B 医学-分析化学
CiteScore
5.60
自引率
3.30%
发文量
306
审稿时长
44 days
期刊介绍: The Journal of Chromatography B publishes papers on developments in separation science relevant to biology and biomedical research including both fundamental advances and applications. Analytical techniques which may be considered include the various facets of chromatography, electrophoresis and related methods, affinity and immunoaffinity-based methodologies, hyphenated and other multi-dimensional techniques, and microanalytical approaches. The journal also considers articles reporting developments in sample preparation, detection techniques including mass spectrometry, and data handling and analysis. Developments related to preparative separations for the isolation and purification of components of biological systems may be published, including chromatographic and electrophoretic methods, affinity separations, field flow fractionation and other preparative approaches. Applications to the analysis of biological systems and samples will be considered when the analytical science contains a significant element of novelty, e.g. a new approach to the separation of a compound, novel combination of analytical techniques, or significantly improved analytical performance.
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