Yanzhen Lu , Yuanyuan Liu , Fenfang Che , Mengyao Gao , Aoxin Li , Yun Wei
{"title":"采用响应面方法优化从 Malus pumila Mill.采用响应面方法对高速逆流色谱法分离叶多酚进行优化。","authors":"Yanzhen Lu , Yuanyuan Liu , Fenfang Che , Mengyao Gao , Aoxin Li , Yun Wei","doi":"10.1016/j.jchromb.2024.124230","DOIUrl":null,"url":null,"abstract":"<div><p>Considering comprehensive utilization of natural products, isolation and activity determination processes of bioactive compounds are essential. In this study, a combined high-speed countercurrent chromatography (HSCCC) with preparative HPLC method was developed to isolate the five antioxidant polyphenols from 75% ethanol extract of <em>Malus pumila</em> Mill. leaves. The HSCCC conditions were optimized by response surface methodology (RSM) considering two response indexes including retention of stationary phase and analysis time. The optimal HSCCC conditions were flow rate of 2.11 mL/min, revolution speed of 717 rpm, and temperature of 25℃, with a solvent system of ethyl acetate/methanol/water (10:1:10, v/v/v). The unseparated fractions obtained from HSCCC were subjected to preparative HPLC for further isolation. As a result, phloridzin (15.3 mg), isoquercitrin (2.1 mg), quercetin 3-<em>O</em>-xyloside (1.9 mg), quercetin-3-<em>O</em>-arabinoside (4.0 mg), and quercitrin (2.0 mg) were isolated from 200.0 mg extracts. The purities of these compounds were all above 92%. Their chemical structures were identified by mass spectrometer and nuclear magnetic resonance. The five isolated compounds were further investigated for their rat hippocampal neuroprotective effects against hydrogen peroxide-induced oxidative stress. No cytotoxicity was observed in all tested concentrations. While all five compounds except phloridzin showed significantly neurogenic activities and neuroprotective effects, especially at the concentration of 0.5 mg/L. These results demonstrate that RSM is a suitable technique for optimisation of HSCCC and the isolated polyphenols can be used as antioxidants in pharmaceutical and food products.</p></div>","PeriodicalId":348,"journal":{"name":"Journal of Chromatography B","volume":null,"pages":null},"PeriodicalIF":2.8000,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Optimisation of isolation of polyphenols from Malus pumila Mill. Leaves by high-speed countercurrent chromatography using response surface methodology\",\"authors\":\"Yanzhen Lu , Yuanyuan Liu , Fenfang Che , Mengyao Gao , Aoxin Li , Yun Wei\",\"doi\":\"10.1016/j.jchromb.2024.124230\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Considering comprehensive utilization of natural products, isolation and activity determination processes of bioactive compounds are essential. In this study, a combined high-speed countercurrent chromatography (HSCCC) with preparative HPLC method was developed to isolate the five antioxidant polyphenols from 75% ethanol extract of <em>Malus pumila</em> Mill. leaves. The HSCCC conditions were optimized by response surface methodology (RSM) considering two response indexes including retention of stationary phase and analysis time. The optimal HSCCC conditions were flow rate of 2.11 mL/min, revolution speed of 717 rpm, and temperature of 25℃, with a solvent system of ethyl acetate/methanol/water (10:1:10, v/v/v). The unseparated fractions obtained from HSCCC were subjected to preparative HPLC for further isolation. As a result, phloridzin (15.3 mg), isoquercitrin (2.1 mg), quercetin 3-<em>O</em>-xyloside (1.9 mg), quercetin-3-<em>O</em>-arabinoside (4.0 mg), and quercitrin (2.0 mg) were isolated from 200.0 mg extracts. The purities of these compounds were all above 92%. Their chemical structures were identified by mass spectrometer and nuclear magnetic resonance. The five isolated compounds were further investigated for their rat hippocampal neuroprotective effects against hydrogen peroxide-induced oxidative stress. No cytotoxicity was observed in all tested concentrations. While all five compounds except phloridzin showed significantly neurogenic activities and neuroprotective effects, especially at the concentration of 0.5 mg/L. These results demonstrate that RSM is a suitable technique for optimisation of HSCCC and the isolated polyphenols can be used as antioxidants in pharmaceutical and food products.</p></div>\",\"PeriodicalId\":348,\"journal\":{\"name\":\"Journal of Chromatography B\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":2.8000,\"publicationDate\":\"2024-07-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Chromatography B\",\"FirstCategoryId\":\"1\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1570023224002393\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Chromatography B","FirstCategoryId":"1","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1570023224002393","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
Optimisation of isolation of polyphenols from Malus pumila Mill. Leaves by high-speed countercurrent chromatography using response surface methodology
Considering comprehensive utilization of natural products, isolation and activity determination processes of bioactive compounds are essential. In this study, a combined high-speed countercurrent chromatography (HSCCC) with preparative HPLC method was developed to isolate the five antioxidant polyphenols from 75% ethanol extract of Malus pumila Mill. leaves. The HSCCC conditions were optimized by response surface methodology (RSM) considering two response indexes including retention of stationary phase and analysis time. The optimal HSCCC conditions were flow rate of 2.11 mL/min, revolution speed of 717 rpm, and temperature of 25℃, with a solvent system of ethyl acetate/methanol/water (10:1:10, v/v/v). The unseparated fractions obtained from HSCCC were subjected to preparative HPLC for further isolation. As a result, phloridzin (15.3 mg), isoquercitrin (2.1 mg), quercetin 3-O-xyloside (1.9 mg), quercetin-3-O-arabinoside (4.0 mg), and quercitrin (2.0 mg) were isolated from 200.0 mg extracts. The purities of these compounds were all above 92%. Their chemical structures were identified by mass spectrometer and nuclear magnetic resonance. The five isolated compounds were further investigated for their rat hippocampal neuroprotective effects against hydrogen peroxide-induced oxidative stress. No cytotoxicity was observed in all tested concentrations. While all five compounds except phloridzin showed significantly neurogenic activities and neuroprotective effects, especially at the concentration of 0.5 mg/L. These results demonstrate that RSM is a suitable technique for optimisation of HSCCC and the isolated polyphenols can be used as antioxidants in pharmaceutical and food products.
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.