芹菜中黄酮类化合物及其生物活性研究进展

芹菜是一种常见的蔬菜,在我国分布广泛.通常含有多种黄酮类物质,包括芹菜素及其糖苷、槲皮素、山奈酚、木犀草素等.本文综述了芹菜中的总黄酮含量与品种,取样部位,生长期等因素的关系.在生物活性方面,芹菜黄酮具有抗氧化、抗癌、抵抗心血管疾病功效,在生殖系统方面有抑制雌激素和孕激素分泌等作用.     

天津和杭州两地55～82岁成年人类黄酮摄入量调查

目的:评价天津和杭州两地55~82岁成年人从蔬菜和水果中摄入的类黄酮总量,为老年人健康饮食提供指导。方法:采用连续5d的24h膳食回顾法,对天津和杭州两地的志愿者进行膳食调查;根据膳食调查的结果,计算类黄酮物质的摄入量。结果 :天津和杭州两地志愿者从蔬菜中摄入的类黄酮总量分别为24.50、47.53mg/d,从水果中摄入的分别为42.71、47.81 mg/d;天津和杭州两地志愿者中,分别有36.5%和49.9%的类黄酮来自蔬菜,63.5%和50.1%的类黄酮来自水果;杨梅黄酮的摄入量最高,其次是槲皮素、芹菜配基、坎二菲醇和木犀草素;膳食调查期间,55~82岁志愿者主要的蔬菜类黄酮来源是大白菜、山药、土豆等,水果类黄酮来源是桔子、苹果、桃等。结论:杭州志愿者从蔬菜和水果中摄入的类黄酮总量均高于天津;天津志愿者从水果中摄入的类黄酮高于蔬菜;杨梅黄酮的摄入量最高。     

基于液质联用和偏最小二乘-判别式鉴别大叶和小叶迷迭香

为了建立一种鉴别大叶和小叶迷迭香的方法,采用液质联用(LC-MS/MS)同时测定迷迭香中木犀草素、咖啡酸、鼠尾草酸、鼠尾草酚、槲皮素、芹菜素、迷迭香酚、泽兰黄酮、香叶木素9种物质含量。检测不同批次大小叶迷迭香9种物质的含量,偏最小二乘-判别分析(partial least squares-discriminate analysis,PLS-DA)分析大小叶迷迭香9种物质含量。结果表明:建立的LC-MS/MS方法线性关系良好,回收率高,符合方法学考察要求。鼠尾草酚、鼠尾草酸、咖啡酸、槲皮素、迷迭香酚、香叶木素、泽兰黄酮、木犀草酸的含量在大叶迷迭香和小叶迷迭香中存在重要差异,可以作为鉴别大叶迷迭香和小叶迷迭香的主要特征标记物。     

6种5,7-二羟基黄酮对巨噬细胞M1极化的影响

目的:探究木犀草素、槲皮素、杨梅素、芹菜素、山柰酚及白杨素这6种黄酮对RAW264.7巨噬细胞极化的抑制活性及构效关系。方法:选择100 ng/mL脂多糖刺激RAW264.7巨噬细胞建立炎症模型;利用噻唑蓝法检测细胞增殖活性;流式细胞术检测细胞表面蛋白CD274和CD38表达;Western blot检测炎性信号通路中相关蛋白的表达;定量聚合酶链式反应检测M1型巨噬细胞标记基因水平。结果:6种黄酮在20 μmol/L下均不影响细胞正常生长;6种黄酮对巨噬细胞CD274和CD38表达均有抑制作用,其中木犀草素、芹菜素和白杨素的抑制作用强于槲皮素、杨梅素和山柰酚;木犀草素对核因子κB信号通路激活的抑制活性强于芹菜素和槲皮素,而杨梅素、山柰酚和白杨素没有明显抑制效果;木犀草素、芹菜素和白杨素抑制iNOS mRNA表达的活性强于其他3种化合物,木犀草素、槲皮素和芹菜素抑制IL-1β及MCP1 mRNA表达的活性强于其他3种化合物。结论:C环上3位的羟基取代不利于黄酮类化合物抑制巨噬细胞M1极化的活性,而B环上3’、4’位羟基取代有利于增强其活性。     

菊花黄酮化合物指纹图谱与抗氧化活性的相关性分析

通过高效液相色谱法方法建立菊花黄酮化合物的指纹图谱，采用灰关联度法和逐步多元回归法对菊花黄酮化合物与其抗氧化活性两者之间的相关性进行分析。结果表明：6种菊花黄酮类化合物的灰关联度大小为槲皮素>木犀草素>杨梅黄素>刺槐素>橙皮素>芹菜素；对菊花黄酮化合物抗氧化活性贡献大小依次为槲皮素>木犀草素>刺槐素>杨梅黄素>芹菜素>橙皮素，其中前4个共有峰与抗氧化活性的灰关联度都大于0.8,可作为影响抗氧化活性的主要因素，而芹菜素无显著性，橙皮素呈负相关。建立的菊花黄酮指纹图谱能很好地表征与其抗氧化作用间的关系。     

不同方法比较黄酮类化合物抗氧化性及其构效关系分析

通过三种化学抗氧化方法(DPPH、FRAP和ORAC法)以及细胞抗氧化方法(CAA法)对芹菜素、山奈酚、木犀草素、槲皮素4种黄酮甙元及它们的糖苷化合物抗氧化活性进行比较研究,结果表明DPPH和FRAP法基本一致,芹菜素、牡荆素和异牡荆素这类C环上无羟基且B环只有一个羟基的黄酮抗氧化性极低;在C环上的羟基相同时,B环有3',4'-邻二羟基的木犀草素和槲皮素的抗氧化活性显著高于仅含4'-OH的芹菜素和山奈酚;而ORAC法的结果与此相反。化学抗氧化测定结果均表明,当C环羟基被取代形成3-O-连黄酮苷时其抗氧化活性减小;而A环上6、8号位的氢被取代形成C-连黄酮苷时其抗氧化性反而会增加。CAA法结果显示,当C环无羟基时,B环只有一个4-OH的芹菜素及其黄酮苷无细胞抗氧化性,但B环有两个羟基的木犀草素及其糖苷却显示出较强的氧化性。与化学抗氧化性不同,A环上C-连黄酮苷的抗氧化性低于其苷元,而C环O-连糖苷的影响则较为复杂。     

异金雀花素等5种大青叶黄酮的同时检测

比较不同来源大青叶提取物对芦丁、槲皮素、木犀草素、山奈酚和异金雀花素5种黄酮物质的高效液相色谱法检测及大青叶来源对其黄酮物质含量的影响。采用DIAMONSIL 5MM C18色谱柱(250 mm×4.6 mm,5μm),流动相A相为0.33%乙酸水溶液;流动相B相为85%甲醇水溶液,梯度洗脱,流速0.80 mL/min,VWD检测波长280 nm;柱温30℃。检测方法显示,5种黄酮物质的检测线性范围为0.02~500 mg/L;检出限(S/N=3)在0.002~0.02 mg/kg;在加标浓度为10.0~90.0 mg/kg条件下,回收率(N=6)在97.99%~103.73%,检测方法灵敏有效。表明不同来源大青叶得到的大青叶提取物中芦丁、槲皮素、木犀草素、山奈酚和异金雀花素5种大青叶提取物黄酮物质的总含量以内蒙古巴彦淖尔产大青叶提取物为最高。     

红甜菜根中类黄酮类差异代谢物的比较分析

该研究以一品红和俄罗斯红甜菜块根为原料,使用UPLC-MS/MS检测平台和Metware自建数据库对两种红甜菜块根中的类黄酮物质进行检测和分析其代谢物质的成分及差异。利用多元统计分析方法,根据检测到的类黄酮类代谢物的数据来筛选差异代谢物;随后再利用KEGG网站分析差异代谢物参与的合成途径。结果表明,两种甜菜块根共检测出类黄酮代谢物69个,分成黄酮、黄酮醇、查尔酮、二氢黄酮、橙酮类和其他类黄酮六种类型。差异代谢物29种,一品红中12种差异代谢物含量高于俄罗斯红,17种低于俄罗斯红。根据代谢物log2FC判定差异倍数较大的成分主要是五羟基查耳酮类、异鼠李素、二甲氧基黄酮、蒽酮、多甲氧基黄酮、槲皮素、山柰酚、香叶木素、高车前素类衍生物。被注释到KEGG 通路上的代谢物有7个：依次是3-O-甲基槲皮素、木犀草甙、3''-O-甲基木犀草素、芦丁、异槲皮苷、异牡荆素、三叶豆甙。由此表明,两种甜菜物质中存在具有显著差异的类黄酮类代谢物,建议可作为特征代谢物做进一步分析。     

冬瓜子提取物中黄酮类物质的同时检测

比较不同来源冬瓜子提取物对芦丁、槲皮素、木犀草素、山奈酚、异鼠李素5种黄酮物质的高效液相色谱法检测及冬瓜子来源对其黄酮物质含量的影响。采用SYMMETRYSHIELD RP18色谱柱(250 mm×4.6 mm,5μm),流动相A为0.3%乙酸水溶液;流动相B为80%甲醇水溶液,梯度洗脱,流速0.80 m L/min,DAD检测波长282 nm;柱温30℃。检测方法显示,5种黄酮物质的检测线性范围为0.05~200 mg/L;检出限(S/N=3)为0.005~0.08 mg/kg,加标回收率(N=6)为87.05%~107.12%,检测方法灵敏有效。检测结果表明,不同来源冬瓜子得到的冬瓜子提取物中芦丁、槲皮素、木犀草素、山奈酚、异鼠李素5种冬瓜子提取物黄酮物质的总含量以河北廊坊产冬瓜子提取物最高。     

芦苇叶类黄酮高效液相色谱分析

目的：分析芦苇叶类黄酮的组分。方法：制备芦苇叶类黄酮提取浓缩过程中形成的析出物和浓缩液再经大孔树脂纯化后产生的初纯物,优化两种样品中类黄酮苷转化为苷元的水解条件,采用高效液相色谱(HPLC)法测定两种样品和其水解样品类黄酮的组分。结果：析出物和初纯物的总黄酮含量分别为57.1%和18.5%,酸浓度1.4mol/L、温度80～90℃条件下水解4h可获得相对最好的水解效果;HPLC测定表明,析出物含32种组分,其中7种得到确定,初纯物含22种组分,其中5种得到确定;水解析出物含28种组分,其中7种得到确定,水解初纯物含25种组分,其中7种得到确定。结论：芦苇叶含有芦丁、野黄芩苷、橙皮苷、木犀草素、槲皮素、芹菜素、山奈酚、异鼠李素(或橙皮素)、异甘草素和黄芩素,其中芹菜素含量最高,还有许多未确定的类黄酮。     

Phenols,proanthocyanidins, flavones and flavonols in some plant materials and their antioxidant activities

Methanol extracts prepared from five plant materials native to the Mediterranean area, namely olive tree (Olea europaea) leaf, St. John's wort (Hypericum perforatum), hawthorn (Crataegus laevigata), oregano (Origanum vulgare) and laurel leaf (Lauris nobilis), were examined for their phenolic components. Total phenolic content was determined by the Folin–Ciocalteu method. The content of proanthocyanidins in acid-hydrolysed extracts was determined spectrophotometrically. The contents of free flavones (apigenin and luteolin) and flavonols (kaempferol, myricetin and quercetin) were determined by HPLC analysis. The time of hydrolysis of flavones, flavonols and proanthocyanidins was optimised.Antioxidant activities of apigenin, luteolin, kaempferol, myricetin, quercetin and of plant extracts were examined. Antioxidative activities were studied in sunflower oil at 98 °C, by measuring peroxide value, and in an aqueous emulsion system of β-carotene and linoleic acid by measuring the absorbance of the sample. Among flavones and flavonols investigated, only myricetin inhibited oxidation of sunflower oil. All other flavones and flavonols showed pro-oxidative activity. Oppositely, in the emulsion system, only apigenin showed pro-oxidative activity while other flavones and flavonols and plant extracts inhibited oxidation of β-carotene.     

不同前处理发酵的桑椹紫酒中黄酮醇物质的比较研究

采用反相高效液相色谱检测不同前处理发酵的桑椹紫酒中黄酮醇的含量,并与葡萄酒中黄酮醇的含量进行对比。结果表明,不同前处理桑椹紫酒中各类黄酮醇的含量除山萘酚,其余均差异极显著。桑椹紫酒中可以检测出10种黄酮醇。其中,槲皮素、杨梅素与葡萄酒中的含量相当,其余8种黄酮醇：山萘酚、异鼠李素、高良姜素、芦丁、非瑟酮、桑色素、鼠李素、芹菜素含量均比葡萄酒中的含量要高,且均在0.54 mg/L以上。     

A critical evaluation on the reliability of two aluminum chloride chelation methods for quantification of flavonoids

Flavonols kaempferol, quercetin, myricetin and gossypetin, and flavones apigenin, acacetin, luteolin, orientin and tricin, are subjected to two AlCl(3) spectrophotometric methods used for determination of total flavonoid content. The method developed by Woisky and Salatino involves addition of AlCl(3) solution to the flavonoid solution and recording of optical density at 420nm. All flavonols except kaempferol have absorption maxima above 440nm and so readings at 420nm are erroneous. Among flavones, all except for luteolin and orientin, have absorption maxima below 400nm. Further, addition of CH(3)COOK and recording the absorbance at 415nm, as modified by Chang et al., works well for flavonols kaempferol, quercetin and myricetin, but not for gossypetin. The flavones luteolin and orientin absorbed above 400nm, whereas all others absorbed below 400nm. Examination of the results of both methods indicates they are inadequate, and should not to be considered as universal and standard methods for total flavonoid determination.     

Determination of flavone C-glycosides in tea

An HPLC method for the determination of flavone C-glycosides (FCG) from black tea has been developed. Sample clean-up was accomplished by means of polyamide column chromatography, followed by enzyme hydrolysis of interfering compounds such was flavonol glycosides and a second polyamide column Chromatographic step. Using HPLC with gradient elution and photodiode array detection eight FCG were separated. Seven FCG were isolated by means of preparative HPLC. Identification was carried out using co-chromatography, FAB(Fast Atom Bombardment)-mass spectrometry and various nuclear magnetic resonance techniques. Apigenin 6-C-glucosyl-8-arabinoside (schaftoside) and apigenin 6-C-arabinosyl-8-C-glucoside (isoschafto-side) as well as luteolin 8-C-glucoside (orientin) and luteolin 6-C-glucoside (isoorientin) have been detected in tea for the first time. Three of the other compounds have been identified as apigenin 8-C-glucoside (vitexin), apigenin 6-C-glucoside (isovitexin) and apigenin 6,8-di-C-glucoside (vicenin-2). Their occurrence in tea has been previously reported. From its UV spectrum another compound was concluded to be an apigenin glycoside. The FCG were quantified in a variety of teas of different origins (16 black, two green and one oolong). The total amounts of the FCG were 0.48–2.69 g/kg dry weight. The FCG pattern of teas of different origins were similar to each other and no origin-dependent characteristics have yet been observed. Small amounts of FCG (1.2–2.2 mg/g) were detected in hydrolysates of high relative molecular mass fractions (Mr>5000) of a black tea liquor.     

Phenolic compound profile of selected vegetables frequently consumed by African Americans in the southeast United States

The phenolic composition of vegetables commonly consumed by African Americans in the southeast United States was analyzed with HPLC–MS. The vegetable samples included collard greens, mustard greens, kale, okra, sweet potato greens, green onion, butter beans, butter peas, purple hull peas, rutabagas, eggplant, and purslane. Five compounds out of total 29 peaks detected from the 12 samples – caffeic acid, ferulic acid, quercetin, kaempferol, and isorhamnetin – were identified. No gallic acid, p-coumaric acid, myricetin, luteolin, apigenin, hesperetin, naringenin, or flavanols was detected. The major flavonoids were isorhamnetin, quercetin and kaempferol. Isorhamnetin was found in kale, mustard greens, and purslane. The content ranged from 2.8 to 23.6 mg/100 g fresh edible part. Quercetin was found in collard greens, mustard greens, kale, okra, sweet potato greens, purple hull peas, and purslane. The content ranged from 1.3 to 31.8 mg/100 g with the highest content in kale and lowest content in purslane. Kaempferol was found in collard greens, mustard greens, kale, sweet potato greens, green onion, and purslane. The content ranged from 1.1 to 90.5 mg/100 g. Caffeic acid was only found in sweet potato greens. Ferulic acid was found in collard greens, mustard greens, kale, okra, purple hull peas, and purslane. Although some peaks were found in eggplant, butter beans, butter peas and rutabagas, these peaks were not identified due to lack of reference compound and no flavonoid or phenolic acid was quantified in these samples. The results suggest that these indigenous vegetables among African Americans are good sources of the phenolic compounds, which can be useful for the prevention of cardiovascular and other chronic diseases.     

Determination of red wine flavonoids by HPLC and effect of aging

A new method for simultaneous determination of 10 flavonols and 2 flavones by high performance liquid chromatography was developed in this paper. The identified compounds contained quercetin, kaempferol, myricetin, rhamnetin, isorhamnetin, quercetrin, rutin, morin, galangin, fisetin, apigenin and luteolin. The chromatographic separation of these flavonoids was performed in a single run by using the mobile phase gradient elution of acetonitrile–methanol–water mixture (1% tetrahydrofuran, THF) at 20 °C, with the flow rate at 1.0 ml/min and the detection wavelength at 360 nm. With direct injection of wine samples, seven red wine samples, differing in their origin of producing places and time, were analyzed for flavonoids content by this method. The results showed the presence of myricetin, luteolin, quercetin, kaempferol, isorhamnetin and galanin. Additionally, the changes of flavonoids in red wines stored in the three types of oak barrels with aging time were investigated, which indicated that the component of flavonoids in red wine is related to wine aging greatly. These provide a substantial basis for the further research on control of flavonoids during winemaking.     

Changes of flavonol synthase and flavonol contents during grape berry development

Flavonols as an important kind of flavonoids not only have many defense-related functions, but also play an important role as potent antioxidants. Flavonol synthase (FLS) as the key enzyme in flavonol synthesis determines the final contents of flavonols in plant materials. The objective of this work was to study the changes of total flavonol content, free flavonol content, the individual flavonol aglycone contents and the two flavones (luteolin and apigenin) contents during grape berry development. The activity and the amount of the changes of FLS during grape berry development were also estimated. The results showed that quercetin, myricetin, kaempferol, isorhamnetin and galangin were detectable during grape berry development, and quercetin aglycone predominated in the total flavonol content in rapid growth phase, lag growth phase and the latter part of the ripening phase, myricetin aglycone witnessed a great increase at veraison, although kaempferol, isorhamnetin and galangin only accounted for a little part of the total flavonol contents but their individual contents all increased after veraison and kaempferol witnessed its highest content at the ripening phase. When it came to the contents of total phenols, total flavonoids, total flavonols and free flavonols, all of them showed accumulated peaks at both the rapid growth phase and the ripening phase. The result of FLS activity had a highly positive correlation with the total contents of flavonols, and the immunoblotting analysis detected two proteins whose signal intensities were in accordance with the FLS activities.     

Identification of phenolic compounds and their relationships to in-vitro digestibility of sorghum leaves from bird-resistant and non-bird-resistant varieties

Soluble phenolics in leaf blades and sheaths from the crop residues of 24 sorghum varieties were studied. Apigenin, luteolin, apigenin and luteolin 7-O-glucosides, p-coumaric acid, butin and apigeninidin were identified. This is the first report of butin in sorghum tissues. Derivatives of the following compounds were also detected but not characterised further: apigeninidin, luteolinidin, chalcone, flavanone and/or dihydroflavonol, cinnamic acid, apigenin and luteolin. The composition of phenolics was clearly different between leaf blades (LB) and sheaths (LS), and also between leaf sheaths of bird-resistant (BR) and non-bird-resistant (NBR) varieties. Routes of flavonoid biosynthesis in BR and NBR varieties appear to diverge at the flavanone/dihydroflavonol level. Several negative correlations were found between HPLC peaks and in-vitro digestibilities, ie true dry matter and neutral-detergent fibre digestibility. These were highly significant with butin and significant with several luteolin derivatives but only with one apigenin derivative. Butin in turn was highly negatively correlated with colorimetric measurements of 3-desoxyanthocyanidins. This may suggest that butin—rather than the 3-desoxyanthocyanidins as previously reported—is implicated in reducing dry matter digestibility. Selecting varieties low in luteolin derivatives and butin and rich in apigenin should enhance the digestibilities of sorghum crop residues.     

Optimization of ultrasonic-assisted enzymatic hydrolysis for the extraction of luteolin and apigenin from celery

The extraction of flavonoids is of increasing interest because of their various pharmacological effects. This study is the first attempt for the ultrasonic-assisted enzymatic hydrolysis (USAEH) applied in the extraction of 2 bioactive flavonoid compounds in celery--luteolin and apigenin. The quantitative yields of luteolin and apigenin were determined by high-performance liquid chromatography (HPLC). To achieve high yields of extracted compounds, the procedure was optimized with regard to the relative parameters involved. The optimal conditions for enzymatic hydrolysis using pectinase treatment were a reaction time of 30 min and a concentration of 0.4 mg/mL at pH 3 for luteolin and pH 5.5 for apigenin. The optimal ultrasonic parameters were an exposure period of 30 min at a temperature of 25 °C using a power source of 80 W. Under these optimal conditions, the yields of luteolin and apigenin were increased to 42.5 and 25.3 mg/g, respectively, which represented a 26.1-fold and a 32.2-fold increase in the yields of these 2 compounds, respectively, compared with the control model of aqueous extraction without enzyme or ultrasonic treatment.     

Metabolomics reveals consistency of the shoot system in Medicago truncatula by HPLC‐UV‐ESI‐MS/MS

Flavonoids not only play crucial roles in plant development and resistance, but also provide one of the major natural sources in human nutrition. To investigate the distribution of flavonoids in the shoot system of Medicago truncatula, a high‐performance liquid chromatography coupled with electrospray ionisation tandem mass spectrometer (HPLC‐ESI‐MS/MS) method was established and then applied to determine the quantitation of flavonoids in different parts of the plant. There were twenty‐two, fifteen and eleven different kinds of flavonoids identified from the flower, leaf and stem of M. truncatula, respectively. The identified constituents were either aglycone or glycosides of the typical flavonoid backbones, such as myricetin, quercetin, luteolin, kaempferol, tricin, apigenin and laricitrin. It was found that the shoot system of M. truncatula can be differentiated by flavonoids in terms of structures and contents. Our results provide instruction to utilise the shoot system of legume crops as fodder and herb medicine in the future.