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基于四丁基溴化铵的疏水型低共熔溶剂用于从藤茶中提取和分离二氢杨梅素及其对黄嘌呤氧化酶的抑制效率

Tetrabutylammonium bromide-based hydrophobic deep eutectic solvent for the extraction and separation of dihydromyricetin from vine tea and its inhibitory efficiency against xanthine oxidase.

作者信息

Wang Liling, Wang Yanbin, Chen Meixu, Zhu Yaoyao, Qin Yuchuan, Zhou Yifeng

机构信息

The Key Laboratory of Biochemical Utilization of Zhejiang Province, Zhejiang Academy of Forestry Hangzhou 310023 China.

School of Biological and Chemical Engineering, Zhejiang University of Science and Technology Hangzhou 310023 China

出版信息

RSC Adv. 2022 Oct 7;12(44):28659-28676. doi: 10.1039/d2ra04266e. eCollection 2022 Oct 4.

Abstract

In this study, deep eutectic solvent oscillation-assisted extraction (DES-OS) combined with macroporous resin adsorption and desorption technology was used to achieve the rapid green extraction and separation of the characteristic component dihydromyricetin (DMY) from vine tea. Multivariate data analysis showed that the DES system composed of tetrabutylammonium bromide (NBr) and pyruvic acid (molar ratio 1 : 2) had good extraction performance for DMY. The influence parameters of DES-OS were studied, and optimized by the single-factor test and response surface methodology (RSM) with Box-Behnken design (BBD). The extraction model of DMY was established and verified. The results showed that the extraction yield of DMY could reach 40.1 mg g under the optimal conditions (DES water contents of 71.18%, extraction time of 2.80 h, extraction temperature of 46.40 °C), which is in good agreement with the predicted value. In addition, Fourier transform infrared spectroscopy (FT-IR) was used to characterize the solvent before and after extraction. Scanning electron microscopy (SEM) results further confirmed that tetrabutylammonium bromide:pyruvate enhanced the destruction of the cell wall structure, resulting in the release of more DMY. Furthermore, different macroporous resins were selected for the separation of DMY for the DES-OS extract, and it was found that the DM301 resin had the ideal recovery performance under optimized dynamic condition. Finally, the product was found to have an inhibitory effect against xanthine oxidase (XO) as a mixed-type competitive inhibitor with IC50 values of (5.79 ± 0.22) × 10 mol L. The inhibitory mechanisms of DMY on XO were explored by enzyme kinetics, spectroscopy, molecular docking and molecular dynamics analysis approaches, which provided a theoretical basis for the above inhibition assays.

摘要

在本研究中,采用深共熔溶剂振荡辅助萃取(DES-OS)结合大孔树脂吸附和解吸技术,实现了从藤茶中快速绿色提取和分离特征成分二氢杨梅素(DMY)。多变量数据分析表明,由溴化四丁铵(NBr)和丙酮酸(摩尔比1∶2)组成的DES体系对DMY具有良好的萃取性能。研究了DES-OS的影响参数,并通过单因素试验和采用Box-Behnken设计(BBD)的响应面法(RSM)进行优化。建立并验证了DMY的萃取模型。结果表明,在最佳条件下(DES含水量71.18%、萃取时间2.80 h、萃取温度46.40℃),DMY的萃取率可达40.1 mg/g,与预测值吻合良好。此外,利用傅里叶变换红外光谱(FT-IR)对萃取前后的溶剂进行了表征。扫描电子显微镜(SEM)结果进一步证实,溴化四丁铵:丙酮酸增强了细胞壁结构的破坏,从而释放出更多的DMY。此外,选择不同的大孔树脂对DES-OS提取物中的DMY进行分离,发现在优化的动态条件下,DM301树脂具有理想的回收性能。最后,发现该产物对黄嘌呤氧化酶(XO)具有抑制作用,作为混合型竞争性抑制剂,IC50值为(5.79±0.22)×10⁻⁵ mol/L。通过酶动力学、光谱学、分子对接和分子动力学分析方法探讨了DMY对XO的抑制机制,为上述抑制试验提供了理论依据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f69/9540247/27176efb1b69/d2ra04266e-f1.jpg

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