Faculty of Food Science and Engineering, Kunming University of Science and Technology (KUST), Kunming 650500, China.
Department of Chemistry, COMSATS University Islamabad, Abbottabad Campus, Abbottabad 22060, Pakistan.
Int J Mol Sci. 2023 Aug 10;24(16):12646. doi: 10.3390/ijms241612646.
The aim of this study was to investigate the chemical composition and antioxidant capacity of various polar fractions obtained from Hook (DH). First, a 90% ethanol-aqueous extract of DH (CF) was subjected to sequential fractionation using different organic solvents, resulting in the isolation of a methylene chloride fraction (DF), an ethyl acetate fraction (EF), an n-butanol fraction (BF), and a remaining water fraction (WF) after condensation. Additionally, the CF was also subjected to column chromatography via a D101 macroreticular resin column, eluted with ethanol-aqueous solution to yield six fractions (0%, 20%, 40%, 60%, 80%, and 100%). UPLC-Q-Exactive Orbitrap-MS/MS analysis identified a total of 47 chemical compounds from these polar fractions, including fatty acids, amino acids, phenolic acids, flavonoids, organic heterocyclic molecules, and aromatic compounds. Moreover, DF, EF, and the 60%, 80%, and 100% ethanol-aqueous fractions had higher total phenol content (TPC) and total flavonoid content (TFC) values and greater 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS-) and 1,1-diphenyl-2-picrylhydrazyl (DPPH)-scavenging abilities. In HO-induced HepG2 cells, the aforementioned fractions could increase the activities of antioxidative enzymes NAD(P)H: quinone oxidoreductase 1 (NQO1), superoxide dismutase (SOD), heme oxygenase-1 (HO-1) and catalase (CAT), stimulate glutathione (GSH) synthesis by increasing the activities of glutamic acid cysteine ligase (GCL) and glutathione synthetase (GS), regulate GSH metabolism by increasing glutathione peroxidase (GSH-Px) and glutathione reductase (GR) activities, and reduce levels of reactive oxygen species (ROS) and malondialdehyde (MDA). Furthermore, the antioxidative stress effect of the DH fractions was found to be positively correlated with the activation of nuclear factor-erythroid 2-related factor 2 (Nrf2) protein and the presence of antioxidative chemical constituents. In conclusion, this study highlights the efficacy of both liquid-liquid extraction and macroporous resin purification techniques in the enrichment of bioactive compounds from natural food resources. The comprehensive analysis of chemical constituents and antioxidant effects of different polar fractions from Hook contributes to the understanding of its potential application in functional foods and nutraceuticals.
本研究旨在探讨不同极性部位的化学成分和抗氧化能力，这些极性部位是从 Hook （DH）中提取得到的。首先，将 DH 的 90%乙醇水溶液（CF）用不同的有机溶剂进行连续分步萃取，得到二氯甲烷部位（DF）、乙酸乙酯部位（EF）、正丁醇部位（BF）和缩合后的剩余水部位（WF）。此外，CF 还通过 D101 大孔树脂柱进行柱层析，用乙醇水溶液洗脱，得到 0%、20%、40%、60%、80%和 100%乙醇水溶液洗脱的 6 个馏分。UPLC-Q-Exactive Orbitrap-MS/MS 分析从这些极性部位中共鉴定出 47 种化合物，包括脂肪酸、氨基酸、酚酸、类黄酮、有机杂环分子和芳香族化合物。此外，DF、EF 和 60%、80%和 100%乙醇水溶液部分的总酚含量（TPC）和总黄酮含量（TFC）较高，2,2'-联氮-双（3-乙基苯并噻唑啉-6-磺酸）二铵盐（ABTS-）和 1,1-二苯基-2-苦基肼（DPPH）清除能力较强。在 HO 诱导的 HepG2 细胞中，上述各部位能提高抗氧化酶 NAD（P）H：醌氧化还原酶 1（NQO1）、超氧化物歧化酶（SOD）、血红素加氧酶-1（HO-1）和过氧化氢酶（CAT）的活性，通过增加谷氨酰半胱氨酸连接酶（GCL）和谷胱甘肽合成酶（GS）的活性来刺激谷胱甘肽（GSH）的合成，通过增加谷胱甘肽过氧化物酶（GSH-Px）和谷胱甘肽还原酶（GR）的活性来调节 GSH 代谢，降低活性氧（ROS）和丙二醛（MDA）的水平。此外，DH 部位的抗氧化应激作用与核因子-红细胞 2 相关因子 2（Nrf2）蛋白的激活和抗氧化化学物质的存在呈正相关。综上所述，本研究突出了液液萃取和大孔树脂纯化技术在从天然食品资源中富集生物活性化合物方面的功效。对不同极性部位的化学成分和抗氧化作用的综合分析，有助于了解其在功能性食品和营养保健品中的潜在应用。
