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灰白小豆对血糖控制的作用灰白小豆，又被称为蚕豆或者芸豆，在中国的饮食中有着悠久的历史。
这种豆类食物不仅味道美味，还具有丰富的营养价值和多种保健功效。
其中，对于血糖控制的作用备受关注。
本文将介绍灰白小豆如何对血糖控制发挥积极作用。
1. 低糖指数的食物灰白小豆是一种低糖指数的食物，这意味着它的碳水化合物被消化和吸收的速度较慢，会导致血糖水平的较低升高。
糖指数是评估食物对血糖影响的指标，食物的糖指数越低，它们对于血糖控制的效果越好。
常食用低糖指数食物有助于维持血糖的稳定，减少葡萄糖的快速释放，有助于防止血糖的突然上升。
2. 富含膳食纤维灰白小豆富含膳食纤维，这也是它对血糖控制有益的一个重要原因。
膳食纤维是一种无法被我们的消化酶分解的碳水化合物，因此当我们食用富含膳食纤维的食物时，它们不会立即被转化为葡萄糖。
相反，膳食纤维会吸引水分，形成胃内的胶体，并延缓食物通过消化系统。
这意味着血糖会缓慢而渐进地释放，从而减慢血糖水平的上升速度。
此外，膳食纤维还有助于增加饱腹感，控制食欲和减少食物的摄入量，进一步维持血糖的平稳。
3. 提供稳定的能量灰白小豆含有丰富的慢性碳水化合物，并且亚洲人民常用其作为主食。
相较于快速释放的碳水化合物，慢性碳水化合物在消化过程中被逐渐分解，能够提供稳定的能量供给。
这样一来，身体能够更有效地利用这些能量，避免能量的突然暴增对血糖水平的不利影响。
此外，稳定的能量来源也有助于减少能量的储存，维持身体的平衡和健康。
4. 抗氧化和抗炎作用灰白小豆含有丰富的抗氧化物质，如维生素C、E和β-胡萝卜素等。
这些抗氧化物质可以帮助减轻氧化应激对身体的伤害，保护胰岛素受损。
此外，灰白小豆还含有一些具有抗炎作用的植物化合物，如黄酮类化合物和异黄酮。
这些化合物能够减少炎症反应，改善胰岛素的敏感性，并进一步促进血糖的控制。
5. 具有调节胰岛素的效果灰白小豆中的一些天然化合物，如异黄酮和皂苷等，据研究显示，具有调节胰岛素和胰岛素受体的作用。
北京大学学报(自然科学版) 第60卷 第1期 2024年1月Acta Scientiarum Naturalium Universitatis Pekinensis, Vol. 60, No. 1 (Jan. 2024)doi: 10.13209/j.0479-8023.2023.024甲醛法处理兰炭废水反应条件的优化王亚俐†柏旭波孙娟娟榆林学院化学与化工学院, 榆林摘要利用甲醛将兰炭废水中的酚类物质转化为酚醛树脂, 以便资源化利用兰炭废水。
通过测定酚醛树脂生成过程中挥发酚、COD、氨氮和油的含量, 调控反应时间、温度以及原料比例, 确定制备酚醛树脂的最优反应条件。
同时, 应用X射线晶体衍射(XRD)和扫描电子显微镜(SEM)等表征技术对生成的酚醛树脂进行理化分析。
研究结果显示, 甲醛法处理兰炭废水制备酚醛树脂的最佳条件为反应温度=90ºC, 反应时间=4h, 甲醛与兰炭废水的体积比=1:40。
关键词兰炭废水; 酚醛树脂; 反应条件优化; 表征Optimization of Reaction Conditions for FormaldehydeTreatment of Semi-coking WastewaterWANG Yali†, BAI Xubo, SUN JuanjuanAbstract Phenolic substances in semi-coking wastewater were converted into phenolic resin by formaldehyde, therefore the resources were utilized. Volatile phenol, COD, ammonia nitrogen and oil in semi-coking wastewater before and after phenolic resin formation were detected to optimize the reaction time, temperature and raw mate- rial ratio, eventually the optimal reaction conditions for phenolic resin preparation were determined. Meanwhile, the physicochemical analysis of phenolic resin materials was carried out by XRD and SEM. The results showed that a volume ratio of 1:40 (formaldehyde vs. semi-coking wastewater), a reaction temperature of 90ºC, and a reaction time of 4 hours were the optimum reaction conditions for semi-coking wastewater treated by formaldehyde.Key words semi-coking wastewater; phenolic resin; reaction condition optimization; characterization兰炭产业的快速发展及其对下游产业链的支撑作用, 使其成为陕北地区经济发展的支柱产业之一。
安徽农学通报，Anhui Agri，Sci，Bull，2022，28（05）不同干燥方式对奇亚籽出油率及油脂品质的影响黄明亚1符洪宇2李维2（1成都大帝汉克生物科技有限公司，四川成都6111302；2四川旅游学院食品学院，四川成都610100）摘要：采用以热风干燥、真空冷冻干燥、微波干燥、烤箱干燥等4种干燥方式对奇亚籽进行干燥处理，并利用索氏提取获得粗油，研究不同干燥方式对奇亚籽出油率及油脂品质的影响。
结果表明：4种干燥方式处理的奇亚籽失水率为2.03%～10.56%，出油率为34.33%～41.68%，其中以真空冷冻干燥的失水率和出油率最高；索氏提取的4种干燥方式获得奇亚籽油的颜色为淡黄色至棕黄色，烤箱烘烤的奇亚籽油透明度最差，微波干燥的色泽最差；4种干燥方式获得奇亚籽油酸价为0.82～3.65mg/g，过氧化值为0.0395～0.0906g/100g，其中真空干燥的酸价和过氧化值均最低；多酚含量为7.68～11.34mg/g，DPPH自由基清除率为49.76%～70.02%，其中真空冷冻干燥的多酚含量和DPPH自由基清除率均最高。
由此可见，真空冷冻干燥处理的出油率及油脂品质优于其他3种干燥方式，且获得的奇亚籽油多酚含量和DPPH清除率均为最好，兼具市场价值和营养价值。
关键词：干燥方式；奇亚籽；出油率中图分类号TS224文献标识码A文章编号1007-7731（2022）05-0157-02奇亚籽（Chia Seed）为薄荷类植物芡欧鼠尾草（Sal⁃via hispanica L.）的种子，该植物最早种植于为墨西哥南部和危地马拉等北美洲地区［1］。
奇亚籽中高品质蛋白质、矿物质、必需氨基酸、维生素等营养物质丰富［2］，含油量在25%～50%，且富含ω-3脂肪酸，其中人体必需脂肪酸α-亚麻酸（ALA）约占总脂肪酸量的56.9%～64.8%，ALA含量在各种植物中是最高的［3］。
近年来，随着人们生活水平的不断提高，营养与健康备受关注，类似奇亚籽这样的新型食品原料的研究逐步增多。
全国中文核心期刊矿业类核心期刊（ＣＡＪ－ＣＤ规范）执行优秀期刊Ｊｒ１瓦订壤羡直嫒蔽化曲中酚失化冬物拍分曷今刊呵毛学锋，陈颖，胡发亭（煤炭科学研究总院北京煤化工研究分院，北京１０００１３）摘要：酚类化合物是煤炭直接液化产物中具有较高附加值的主要含氧化合物，也是煤液化油酸性物质的主要成分。
它不仅对油品的提质加工的技术路线有影响，而且也对煤炭直接液化的经济性具有较大的影响。
由于该方面的研究工作尚属起步阶段，借鉴煤焦油和石油系产品中酚类化合物的有关分离与利用情况，结合笔者的研究实践和认识，提出了在煤直接液化油中酚类的分离与利用方面应开展的工作和研究方向。
关键词：直接液化；煤液化油；酚类化舍物；分离与利用中图分类号：ＴＱ５２９．１文献标识码：Ａ文章编号：１００６－６７７２（２００８）０６－００３９—００煤炭直接液化（以下简称“煤液化”）是把固体状态的煤在高压和一定温度下直接与氢气反应，使煤炭直接转化成液体油品的工艺技术¨Ｊ。
煤液化得到的一次粗油中含有较多的含氧化合物，其中大部分足酚类化合物。
目前围内外对煤液化油采用的提质加工方式类似于石油的后续加工方式，即通过二次加氢将其转变为各种规格的成品燃料油。
酚类化合物虽然对煤液化油的后续加工和液化油品的安定性均有一定的影响旧ｊ，但它也是煤液化油中一项具有含量高、市场需求前景广阔的高附加值产品。
随着可预期的煤液化大规模工业化时代来临∞Ｊ，开展对煤液化油中酚类化合物分离与利用研究，具有非常重要的经济效益和社会效益。
１煤液化油组成及特点煤液化油组成结构非常复杂，含有大量芳香烃以及氮、氧和硫等杂原子，且其组成因所用煤种和液化条件而异ＨＪ。
煤液化产物中的芳烃化合物大都带有烷基侧链，主要有烷基萘、烷基菲、烷基蒽等芳烃化合物。
液化油中的碱性组分组成结构比较复杂，主要是含喹啉、八氢啡啶的同系物；煤液化油中的酸性组分则主要足含有各种单体的一元或多元环酚类化合物，如苯酚、二甲基酚、三甲基酚及萘酚，茚酚等含氧化合物。
收稿日期:2013-11-05作者简介:张明成(1982-),男,山东招远人,绥化学院食品与制药工程学院助理实验师,研究方向:农产品加工及贮藏工程。
项目基金:绥化学院青年基金(科学技术)项目(项目编号:KQ1302002)。
∗∗∗第34卷第5期绥化学院学报2014年5月Vol.34No.5Journal of S uihua UniversityMay .2014大豆蛋白中7S 与11S 组分的分离方法摘要:7S 和11S 是大豆蛋白中两种主要组分,由于二者结构不同,因此具备不同的功能性质,如何从大豆蛋白中将这两种组分高效分离是研究者们关注的问题。
文章评述了碱溶酸提法,冷沉法、盐析法等分离方法的发展和不断优化,为不断进步的分离技术提供了参考。
关键词:11S 组分;7S 组分;碱溶酸提法;冷沉法中图分类号:T5201.1文献标识码:A 文章编号:2095-0438(2014)05-0150-04一、前言大豆中含有大量的贮存蛋白质,其含量可高达40%。
根据沉降系数的不同,大豆蛋白可分为2S 、7S 、11S 和15S 四种组分,其中,7S 组分中的β-伴大豆球蛋白(β-co ng lycinin )和11S 组分中的大豆球蛋白(g lycinin )是大豆分离蛋白的主要成分。
由于7S 和11S 氨基酸的组成、亚基的结构及其相互作用不同,他们表现出的功能性质如乳化性、凝胶型等存在较大差异,与11S 球蛋白相比,β-伴大豆球蛋白蛋白含有的巯基和二硫键较少,导致其凝胶保水性和黏结性较差,而含有的赖氨酸和疏水性氨基酸较多导致其具有较强的表面活性、较好的溶解性、乳化性与稳定性[1][2]。
段春红[3]等研究大豆分离蛋白亚基及7S /11S 比例对肉肠品质的影响时发现7S /11S 与肉肠的弹性呈现极显著的正相关性,与咀嚼性和凝胶强度均呈现显著的正相关性,但与硬度的相关性不显著,与得率呈现显著的负相关性。
20世纪50年代起至今,人们一直不断研究7S 与11S 的分离方法,这些方法主要包括碱溶酸提法、冷沉法、盐析法等,其中碱溶酸提法由于其较好的得率和提纯纯度一直被视为经典的分离方法,但近来也有学者在冷沉的基础上研究冻融张明成(绥化学院黑龙江绥化152061)150. All Rights Reserved.处理的分离方法。
气相色谱-质谱法测定白酒中挥发性酚类物质张倩【摘要】用液液萃取作为前处理方法,建立气相色谱-质谱联用法测定白酒中10种挥发性酚类物质含量的方法.该方法检测得出的质谱图在谱库进行检索,准确地对10种挥发性酚类物质进行定性、定量分析.加标回收率为83.9%~101.2%,RSD值为1.1%~2.9%.定性、定量准确度较高,重复性好.实验结果表明,该方法适用于白酒中挥发性酚类化合物的研究.%GC-MS was used to detect the content of 10 kinds of volatile phenolic compounds in Baijiu pretreated by liquid-liquid extraction. Library searching of the obtained mass spectra was performed to achieve more accurate quantitative and qualitative analysis of the 10 volatile phenolic compounds. The recovery rate was 83.9%~101.2%, and RSD was 1.1%~2.9%. Qualitative and quantitative accuracy was high with satisfactory repeatability. The experimental results proved that the method was suitable for the study of volatile phenol compounds in Bai-jiu.【期刊名称】《酿酒科技》【年(卷),期】2016(000)009【总页数】3页(P128-130)【关键词】白酒;挥发性酚;GC-MS【作者】张倩【作者单位】国家酒类及饮料产品质量监督检验中心,贵州贵阳550016【正文语种】中文【中图分类】TS262.3;TS261.7挥发性酚类物质是白酒中的重要香味物质，这一类物质对于酒的闻香、口味以及稳定性等方面均具有重要的作用。
中药锁阳指纹图谱和原花青素的提取及提取物抗氧化的研究高乃群S080106018一.前言在新世纪,具有我国传统文化和独特优势的中药正面着前所未有的挑战和发展机遇.一方面,随着社会的发展,人类疾病谱已悄然发生改变,医疗模式已由单纯的疾病治疗变为预防.保健.治疗.康复相结合的模式.传统医学发挥着越来越大的作用.生存环境的不断恶化,使人类回归自然的忽视越来越高,因而传统医药备受青睐.[2]另一方面,我国传统中药在国际市场正面临诸如日本.韩国.印度泰国等传统医药国家和德国.法国等欧洲国家植物药的巨大冲击.由于我国传统中药生产的众多产品不能符合国际标准和要求,目前只有百分之几的国际市场销售份额也可能进一步萎缩.[3]所以建立一套完整的符合国际标准的中药检测质量体系很重要.HPLC和EC是面前最常用的两种中药的检测标准.中药指纹图谱库的国家标准的建立也势在必行.中药有效成分提取量化也迫在眉睫.中药有效成分的提取的目的是拿到能够实现该药理功能的有效成分，而非本中药的主要成分。
通过定向提取的成分集合，可以用HPLC测得有效成分的含量，根据含量和工艺收得率及原处方的生药用量，计算出该提取物的配伍剂量。
从而使复方的成分得以量化，质量得以稳定。
确保了医生开什么药，患者用什么药;医生开多少，患者用多少;医生怎么开，患者怎么用。
确保了医和药的统一，处方和药品的统一，药性和疗效的统一，医生和患者的统一。
[4][5]二.锁阳中原花青素的研究[原植物〕多年生寄生革伞．无叶绿素．高10一100cm。
茎圆柱状．暗紫红色．有散生鳞片，基部膨大。
穗状花序生于茎顶，棒状、长圆形或狭椭圆形．生密集的花和鳞片状色片,花杂性．暗紫色。
坚果球形。
药材性状〕个品呈扁圆柱形．微弯曲．[1][药材性状]本品为扁圆柱型,长5—15cm。
直径1.5---5 cm表面棕色或棕褐色，组糙．具明显纵沟及不规则凹陷，有的残存三角形的黑棕色鳞片。
体重．质硬．难折断．断面浅棕色或棕褐色．有黄色三角状维管束。
芋头可溶性酚类物质及其与芋头疫病抗性关系公司。
甲醇、乙酸、乙腈购自国药集团化学试剂有限公司（均为色谱纯级）。
水为娃哈哈牌纯净水。
1.2 仪器设备Agilent 1200高效液相色谱仪;真空干燥箱（上海之信仪器有限公司）。
1.3 方法1.3.1 芋头的疫病抗病性鉴定芋头疫病抗性参考Brooks [2]的方法进行，稍作改进。
培养3～5d的疫霉菌打出直径为0.5cm的菌块。
接种于第2或第3最嫩的叶片的背面，用透明胶封住菌块，接种5d后，测定病斑的直径，作为芋头抗疫霉菌的标准。
每个材料接种3张叶片。
抗病鉴定于2016年在广东海洋大学农学院实验基地进行。
1.3.2 标准储备液称取色谱纯级别的没食子酸0.05g，置于10mL的容量瓶中，加入甲醇（色谱纯）溶解，定容。
浓度为5000mg/L。
然后经0.2μm孔径的滤膜过滤后置于4℃冰箱中避光保存，作为贮备液;使用时用甲醇（色谱纯）稀释500倍。
终浓度为10μg/mL。
阿魏酸、白藜芦醇、对香豆酸、绿原酸、芦丁、杨梅素、肉桂酸和鞣花酸的配制方法同没食子酸，终浓度为100μg/mL。
1.3.3 样品前处理选芋头最嫩叶片0.5g，加液氮充分研磨，按De Ascensao的方法提取可溶性酚类物质[5]。
1.3.4 色谱条件色谱柱：Agilent XDB-C 18 柱（4.6mm×150mm，5μm）;流动相：乙腈与10%乙酸溶液，梯度洗脱：0 min → 10min→20min→30min → 40min→55min，乙腈35% →40%→ 55% → 80%→0;检测波长：280 nm;流速：1.0 mL/min;柱温：30℃;进样量：10μL。
2 结果与分析2.1 芋头的疫病抗病性分离出疫霉菌后，经形态学观察及分子鉴定确定为疫霉菌，以此菌作为接种的病原菌。
接种后测定病斑的直径，以其病斑直径的大小作为衡量芋头抗病性的指标。
通过方差分析，差异极显著，表明不同芋头品种之间的抗病性存在极显著差异。
外文翻译（原文）Catalytic wet peroxide oxidation of azo dye (Congo red) using modified Y zeolite as catalystAbstractThe present study explores the degradation of azo dye (Congo red) by catalytic wet peroxide oxidation using Fe exchanged commercial Y zeolite as a catalyst. The effects of various operating parameters like temperature, initial pH, hydrogen peroxide concentration and catalyst loading on the removal of dye,color and COD from an aqueous solution were studied at atmospheric pressure. The percent removals of dye, color and COD at optimum pH07, 90◦C using 0.6 ml H 2 O2/350 ml solution and 1 g/l catalyst was 97% (in 4 h), 100% (in 45 min) and 58% (in 4 h), respectively. The % dye removal has been found to be less in comparison to % color removal at all conditions, e.g. dye removal in 45 min and at above conditions was 82%, whereas the color removal was 100%. The results indicate that the Fe exchanged Y zeolite is a promising catalyst for dye removal. Fe exchanged catalyst is characterized using XRD, SEM/EDAX, surface area analyzer and FTIR. Though the dye, color and COD removals were maximum at pH02 but as the leaching of Fe from the catalyst was more in acidic pH range, pH0 7 was taken as operating pH due to almost comparable removals as of pH0 2 and no leaching of Fe ions.© 2008 Elsevier B.V. All rights reserved.1. IntroductionReactive azo dyes from textile and dyeing industries pose grave environmental problem. An estimate shows that textiles account for 14% of India’s industrial production and around 27% of its export earnings[1]. Production during 2006 registered a growth of about 3.5% at 29,500 tonnes and the textile industry accounts for the largest consumption of dyestuffs at nearly 80% [2]. The waste containing these azo dyes is non-degradable. The process of dyeing is a combination of bleaching and coloring, which generates huge quantities of wastewaters causing environmental problems. The effluents from these industries consist of large quantities of sodium, chloride, sulphate, hardness, carcinogenic dye ingredients and total dissolved solids with very high BOD and COD values over 1500 mg/l and over 5000 mg/l, respectively [3]. Various methods have been used for dye removal like adsorption, coagulation, electrocoagulation, Fenton’s reagent and combination of these processes. Though these treatment processes are efficient in dye removal, they generate adsorbed waste/sludge, etc. which further causes a secondary pollution. In wet oxidation the sludge is disposed off to a great extent by oxidizing the organic pollutant. Catalytic wet oxidation method (CWAO and CWPO) is gaining more popularity. CWPO process using H2O2, in particular has advantages like better oxidation ability thanusing oxygen,as the former is carried out at lower pressure (atmospheric pres-sure).WAO usually acts under high temperatures (200–325◦C)and pressure (50–150 bar). A comparable oxidation efficiency is obtained at a less temperature of 100–120◦C when using hydrogen peroxide as the oxi dizing agent instead of oxygen [4].WAO is capital intensive whereas WPO needs limited capital but generates little higher running costs [4].Rivas et al.[5] showed that the addition of H2O2(as a source of free radicals) enhanced wet air oxidation of phenol, a highly non-degradable substance and found that the combined addition of H2O2 and a bivalent metal (i.e. Cu, Co or Mn) enhanced the rate of phenol removal. Various oxidation catalysts have been studied for the removal of different compounds like phenol, benzoic acid, dyes, etc. by CWPO process. Catalysts like Fe2O3/CeO2and WO3/CeO2 in the removal of phenolic solution, (Al–Fe) pillared clay named FAZA in the removal of 4-hydroxy benzoic acid, mixed (Al–Fe) pillared clays in the removal of organic compounds have been used[6–8] .Removal of dyes by CWPO process is gaining importance in recent times with a large number of catalysts. Kim and Lee [9] used Cu/Al2O3 and copper plate in treatment of dye house effluents. Liu and Sun [10] removed acid orange 52, acid orange 7 and reactive black 5 using CeO2doped Fe2O3/ -Al2O3 from dye waste water. Kim and Lee [11] reported the treatment of reactive dye solutions by using Al–Cu pillared clays as catalyst.Among these catalysts, modified zeolites are preferred for improved efficiency, lower by-product formation and less severe experimental conditions (temperatures and pressures). Theimproved efficiency of the catalyst is ascribed to its structure and large surface area with the ability of forming complex compounds. Zeolites can be ion exchanged using transition metal ions like Fe,Cu, Mn and others like Ca, Ba, etc. Zeolites are negatively charged because of the substitution of Si(IV) by Al(III) in the tetrahedral accounts for a negative charge of the structure and hence the Si/Al ratio determines the properties of zeolites like ion exchange capacity [12] . These metal ions neutralizethe negative charge on zeolites and their position, size and number determine the properties of zeolite. These metal ions are fixed to the rigid zeolite framework which prevents leaching and precipitation in various reactions[13–21] .In this work, catalytic wet peroxide oxidation of Congo red azo dye using Fe exchanged Y zeolite has been presented. Effect of variables like temperature, initial pH, peroxide concentration and catalyst loading on catalytic wet peroxide oxidation were examined and the optimum conditions evaluated.2.Materials and methods2.1. ChemicalsHydrogen peroxide (30% analytical grade), manganese dioxide,sodium hydroxide pellets (AR) and hydrochloric acid were obtained from RFCL limited (Mumbai), India. Congo red was obtained from Loba Chemie Pvt. Ltd. (Mumbai) and were obtained from RFCL limited (Mumbai), India.Commercial Na–Y zeolite was obtained from Sud chemie Pvt.Ltd. (Baroda), India. Commercial catalyst was iron exchanged with excess 1 M Fe(NO3)3 at 80◦C for 6 h. The process was repeated three times and the sample was thoroughly washed with distilled water and dried in oven in air at60◦C for 10-12 h. The amount of iron exchanged was 1.53 wt% estimated by A.A.S.2.2. Apparatus and procedureThe experimental studies were carried out in a 0.5 l three-necked glass reactor equipped with a magnetic stirrer with heater and a total reflux (Fig. 13). Water containing Congo red dye was transferred to the three-necked glass reactor. Thereafter, the catalyst was added to the solution. The temperature of the reaction mixture was raised using heater to the desired value and maintained by a P.I.D. temperature controller, which was fitted in one of the necks through the thermocouple. The raising of the temperature of the reaction mixture to 90◦C from ambient took about 30 min.The total reflux prevents any loss of vapor and magnetic stirrer to agitate the mixture. Hydrogen peroxide was added, the runs were conducted at 90◦C and the samples were taken at periodic intervals. The samples after collection were raised to pH-11 by adding 0.1N NaOH (so that no further reaction takes place) and the residual hydrogen peroxide was removed by adding MnO2 which catalyzed the decomposition of peroxide to water and oxygen. The samples were allowed to settle for overnight or one day (or centrifuged) and filtered. The supernatant was tested for color and COD. After the completion of the run, the mixture was allowed to cool and settle overnight.2.3. CharacterizationThe determination of structure of the heterogeneous catalyst was done by X-ray diffractometer (Bruker AXS, Diffraktometer D8,Germany). The catalyst structure was confirm ed by using Cu Kα as a source and Ni as a filter. Goniometer speed was kept at 1cm/min and the chart speed was 1 cm/min. The range of scanning angle(2θ) was kept at 3–60◦. The intensity peaks indicate the values of2θ , where Bragg’s law is applicable. The formation of compounds was tested by comparing the XRD patternusing JCPDS files (1971).The determination of images and composition of catalyst were done by SEM/EDAX QUANTA 200 FEG. Scanning for zeolite samples was taken at various magnifications and voltage to account for the crystal structure and size. From EDAX, the composition of the elements in weight percentage and atomic percentage were obtained along with the spectra for overall compositions and particular local area compositions. BET surface area of the samples was analyzed by Micromeritics CHEMISORB 2720. The FTIR spectra of the catalyst was recorded on a FTIR Spectrometer (Thermo Nicolet, USA, Software used: NEXUS) in the 4000–480 cm−1wave number range using KBr pellets. The internal tetrahedra and external linkage of the zeolites formed are identified and confirmed by FTIR. The IR spectra data in Table 2 is taken from literature[22] .2.4. AnalysisThe amount of the dye present in the solution was analyzed by direct reading TVS 25 (A) Visible Spectrophotometer. The visible range absorbance at the characteristic wavelength of the sample at 497 nm was recorded to follow the progress of decolorization during wet peroxide oxidation.The COD of the dye solution was estimated by the Standard Dichromator Closed Reflux Method (APHA-1989) using a COD analyzer (Aqualytic, Germany). The color in Pt–Co unit was estimated using a color meter (Hanna HI93727, Hanna Instruments, Singapore) at 470 nm and the pH was measured using a Thermo Orion, USA make pH meter. The treated dye solutions were centrifuged (Model R24, Remi Instruments Pvt. Ltd., Mumbai, India) to obtain the supernatant free of solid MnO2.A.A.S (Avanta GBC, Australia) was used to find the amount of iron exchanged and leached.3. Results and discussionDue to the iron present after the exchange process, the Y peaks diminished along with the rise in Fe peaks. Similar phenomena has also been observed by Yee and Yaacob [23] who obtained zeolite iron oxide by adding NaOH and H2O2(drop wise) at 60◦C to Na–Y zeolite. XRD pattern ( Fig. 2) showed diminishing zeolite peaks along with evolution of peaks corresponding to y-Fe2O3 with increasing NaOH concentration. The IR assignments from FTIR (Fig. 3) remain satisfied even after iron exchanging. The EDAX data (Table 1) show clearly an increase in the value of Fe conc. after ion exchange of Y-zeolite. The BET surface area (Table 1) has been found to decrease from 433 to 423 m2/g after Fe exchange. SEM image is shown in Fig. 1 . Table 2 presents FTIR specifications of zeolites (common to all zeolites).The effect of temperature, initial pH, hydrogen peroxide concentration and catalyst loading on catalytic wet peroxide oxidation of azo dye Congo red were investigated in detail.Fig. 1. SEM image of Fe-exchanged Y zeolite.Fig. 2. XRD of commercial and Fe-exchanged commercial Y zeolite.BET surface area (commercial Na–Y): 433.4 m2/g.BET surface area (Fe exchanged commercial Na–Y): 423 m2/g.Table 2Zeolite IR assignments (common for all zeolites) from FTIR.3.1. Effect of temperature on dye, color and COD removalThe temperatures during the experiments were varied from50◦Cto100◦C. A maximum conversion of dye of 99.1% was observed at 100◦C in 4 h (and 97% at 90◦C). The dye rem ovals at 80◦C, 70◦C, 60◦C and 50◦C and at 4 h are 56%, 52%, 42% and 30%,respectively. Fig. 4 shows that at a particular temperature, the dye concentration gradually decreases with time. The initial red color of the dye solution decreased into brown color in due course and finally the brown color disappeared into a colorless solution. Dye concentration decreases at faster rates with temperatures for initial 30 min and thereafter it decreases from 1 h to 2 h. The initial concentrations of dye did not change after a brief contact period of dye solution with the Fe-exchanged zeolite catalyst (before CWPO)confirming that there is negligible adsorption of the dye by the catalyst.Fig. 5 shows the results obtained for color removal as a function of time and temperature. The maximum color removal (100%) is obtained at 100◦C in 30 min and also at 90◦C in 45 min. At a particular temperature, the color continuously decreases with time atFig. 3. FTIR of Fe-exchanged Y zeolite.Fig. 4. % dye removal as function of temperature.faster rate in first few minutes until a certain point ( t = 45 min) and then remaining almost unchanged. At 50◦C, the color removal is very low, whereas at 60◦C, there is a sudden shift towards its greater removal. The color removal is much higher at higher temperatures(70–100◦C).Fig. 6 depicts the results obtained for %COD removal as a function of time and temperature. A maximum COD removal of 66% was obtained at 100◦C (at 4 h) followed by 58% at 90◦C (at 4 h). Until60◦C, the rate of COD removal is less and during 70–100◦C, the rate is much faster.3.2. Effect of initial pH on dye, color and COD removalThe influence of initial pH on dye (Congo red) removal was studied at different pH (pH0 2, 4, 7, 8, 9 and 11) without any adjustment of pH during the experiments. A maximum conversion of 99% was obtained at pH0 2 followed by 97% at pH0 7. The dye removal at pH0 4, 8, 9 and 11 were 94%, 29%, 5% and 0.6%, respectively. All the runs were conducted for 4 h duration. The color of the solution is violet blue at pH0 2 (a colloidal solution) and greenish blue at pH0 4 (colloidal solution). In neutral and basic pH0(7, 8, 9 and 11) range, color of the solution did not change during treatment and was same as original solution, i.e. red color. Fe cations can leach out from zeolite structure into the solution causing secondary pollution. Leaching of Fe cations out of zeolitesFig. 5. % color removal as function of temperature.Fig. 6. %COD removal as function of temperature.Fig. 7. % color removal as function of pH0depends strongly on pH of the solution. The leaching of iron ions was enhanced at low pH values [24,25] . In order to determine dissolved Fe concentration, final pH values of the solutions were analyzed by A.A.S. At initial pH0 2 and 4, Fe detected in the solution was 7.8 ppm and 3.9 ppm, respectively. At pH0 7 and in alkaline range, there wasFig. 8. %COD removal as function of pH0.Fig. 9. % color removal as function of peroxide concentration.Fig. 10. %COD removal as function of peroxide concentration.almost no leaching. pH0 7, therefore, was chosen to be optimum pH for future experiments. The final pH values pH f after the reaction corresponding to pH0 2, 4, 6, 8, 9 and 11 were 2.1, 4.2, 7.2, 7.7 and 8.7, respectively. This show that the pH f tend to reach to neutral pH for all starting pH values.Fig. 7 presents the results obtained for color removal as a function of time and pH0. A maximum color removal of 100% was obtained at pH0 2 (in 10 min) and also at pH0 7 (in 45 min). The color removal at a particular pH0 decreases at a faster rateinitially (0–1 h) and thereafter it has a slower rate. The lowest removal was observed at pH0 11 with almost no removal.Fig. 11. % color removal as function of catalyst loading.Fig. 12. %COD removal as function of catalyst loading.The results obtained for COD removal as a function of time and pH0 are shown in Fig. 8 . A maximum COD removal of 69% was obtained at pH0 2 in 4 h followed by 63% at pH0 4 and 58% at pH0 7in4h.Fig. 8 shows maximum decrease in COD value in the initial 30 mines at all pH0. The decrease in COD is not appreciable thereafter. The COD removal is more in acidic range with a maximum removal of 69%, moderate in neutral region and least in basic region.3.3. Effect of peroxide concentration on dye, color and COD removalThe influence of H2O2 concentration on dye removal was investigated at different concentrations of hydrogen peroxide (in the range 0–6 ml). A maximum removal of 99.02% was obtained at H2O2 concentration of 3 ml per 350 ml of solution, followed by 98.3% at 1ml and 97% at 0.6 ml. The dye removal at H2O2concentrations of 6 ml,0.3 ml and 0 ml (and at 4 h) were 94%, 82% and 8%, respectively. The dye removal rate at 90◦C temperature is gradual at all conc entrations of peroxide. At peroxide concentration of 0 ml, there is very little removal of dye, hardly 8%. Hence, it can be inferred that catalytic thermolysis (a process of effluent treatment by heating the effluent with/without catalyst) is not active and cannot be applied for dye removal.At the beginning of the reaction, the OH•radicals which are produced additionally when peroxide concentration is increased,speeds up the azo dye degradation. After a particular peroxide concentration, on further increase of the peroxide, the dye removal isFig. 13. Schematic diagram of the reactor.not increased. This may be because of the presence of excess peroxide concentration, hydroperoxyl radicals (HO2•) are produced from hydroxyl radicals that are already formed. The hydroperoxyl radicals do not contribute to the oxidative degradation of the organic substrate and are much less reactive. The degradation of the organic substrate occurs only by reaction with HO•[26] .The % color removal at a particular peroxide concentration increases at a faster rate in the initial 45 min and then at slower rates afterwards (Fig. 9). As H2O2 concentration increases, the rate of removal is much faster, reaching 100% in 45 minusing 6 ml H2O2 per 350 ml solution, whereas it is 100% in 1 h for both 0.3 ml and3ml.Fig. 10 shows the results obtained for COD removal as a function of time and H2O2 concentration. The maximum COD removal, 63% is obtained for H2O2 conc. 3 ml at 90◦C, pH0 7 and 2 h duration.3.4. Effect of catalyst loading on dye, color and COD removalThe influence of catalyst concentration on dye removal was investigated at different concentrations (in the range 0.5–1.5 g/l). A maximum dye removal of 98.6% was observed at 1.5 g/l followed by 98.3% at 1 g/l and 87.3% at 0.5 g/l in 4 h duration. The % dye removal without catalyst was very low with only 36% dye removal in 4 h. By comparing the results for the dye removal without catalyst and1.5 g/l catalyst, the removal for 1.5 g/l is approximately three times to that of without catalyst. The rate of removal is also more for higher concentrations of catalyst and increases with it.Fig. 11 shows the results obtained for color removal as a function of time and catalyst concentration. The maximum color removal of 100% was obtained using 1.5 g/l catalyst conc. in 1.5 h and also using 1 g/l catalyst in 3 h.Fig. 12 presents the results obtained for %COD removal as a function of time and catalyst concentration. A maximum COD removal of 58% was obtained at catalyst conc. 1 g/l, 51.8% at 1.5 g/l and 50.5% at 0.5 g/l in 4 h. Without catalyst, the COD removal was only 35%.4. ConclusionsThe % removals of dye, color and COD by catalytic wet peroxide oxidation obtained at 100◦C, 4 h duration using 0.6 ml H2O2/350 ml solution, 1 g/l Fe–Y catalyst and pH0 7 were 99.1%, 100% (30 min)and 66%, respectively. As at 100◦C the solution has tendency to vaporize during the operation, 90◦C was taken as operating temperature. The corresponding % removals at 90◦C were 97% dy e, 100%color (in 45 min) and 58% COD. Acidic range gave higher % removals in comparison to neutral and alkaline range. At pH0 2, the dye, color and COD removals of 99%,100% (in 10 min) and 69% were observed after 4 h duration. As at pH0 2, the leaching of Fe ions from Y zeolite catalyst is predominant,pH0 7 was taken as operating pH. Fe concentration of 7.8 ppm was observed in the solution at pH0 2. The values of removals, however,are comparable to pH0 2, with dye removal of 97%, color removal of100% (in 45 min) and COD removal of 58% in 4 h.The H2O2concentration was found to be optimum at 3 ml/350 ml solution giving dye, color and COD removals of 99%,100% (in 1 h) and 63%, respectively.The study on the effect of catalyst loading revealed 1.5 g/l as best among the catalyst concentrations studied. The results with 1 g/l and 1.5 g/l catalyst concentration were almost comparable.外文翻译（译文）使用改性Y沸石为催化剂湿式催化过氧化氢氧化偶氮染料(刚果红)摘要本研究主要探讨了使用改性Y沸石固载铁离子作为催化剂湿式催化过氧化氢氧化降解偶氮染料(刚果红)。
Characterization of phenolic compounds in Erigeron breviscapus by liquid chromatography coupled to electrospray ionization mass spectrometryYufeng Zhang,Peiying Shi,Haibin Qu*and Yiyu Cheng*Pharmaceutical Informatics Institute,Zhejiang University,Hangzhou310027,P.R.ChinaReceived18May2007;Revised29June2007;Accepted30June2007Erigeron breviscapus is an important herbal drug for the treatment of cardiovascular and cerebral vessel diseases.In this study,phenolic compounds were extracted from the whole plant ofE.breviscapus and analyzed by high-performance liquid chromatography/electrospray ionizationmass spectrometry.A total of53compounds were identified or tentatively characterized based on their UV and mass spectra.These compounds included caffeoylquinic acids(CQAs),CQA glucosides,malonyl-CQAs,acetyl-CQAs,caffeoyl-2,7-anhydro-3-deoxy-2-octulopyranosonic acids (CDOAs),caffeoyl-2,7-anhydro-2-octulopyranosonic acids(COAs),flavones,flavonols andflavo-nones.Most of them were reported for thefirst time from E.breviscapus and nineteen of them belonged to new compounds.The MS n spectra of CQA glucosides were similar to CQAs and they were discriminated by their retention times.No caffeic acid related ions(X S0,Y S0and Z S0)were observed in MS n spectra of acyl-CQAs compared to those of CQAs.Fragment ions(2,5O S,3,6O S and 4,6O S)corresponding to ring cleavage were shown in MS n spectra of CDOAs and COAs,character-istic of this class of compounds.The5,6,7-trihydroxyl-substitutedflavones were dominant inE.breviscapus.Their[A–H]S ions underwent the loss of a molecule of H2O,followed by the lossof CO,which was used to discriminate from other hydroxyl-substitutedflavones.Our results are the first comprehensive analysis of E.breviscapus constituents and will be helpful for the quality control of the herb of E.breviscapus and its related preparations.Copyright#2007John Wiley&Sons,Ltd.The whole plant of Erigeron breviscapus(Compositae),also known as Dengzhanxixin,is an important herbal drug in China.Before it was introduced as a specific therapy for hemiplegia by a botanic physician in1960s,this herb had only been used among the minority ethnic groups(such as Miao,Yi and Deang)in southern China.1Since then, extensive studies about its components,pharmacological effects and clinical applications have been carried out.2–4 Moreover,a GAP base of E.breviscapus has been established in Yunnan Province to guarantee the genuine and reasonable exploitation of this herb.According to Chinese medical science,E.breviscapus can promote blood circulation,remove blood stasis,and activate meridians to stop pain.5When applied in clinical practice, this herbal drug showed significant curative effects to cardiovascular and cerebral vessel diseases.6Furthermore, other pharmacological activities,such as neuroprotective,7 antioxidant,8antibacterial and antifungal activity,9were also discovered.The main active components identified from this herb includedflavonoids,coumarins,lignins,hydroxy-cinnamic acids,pyromeconic acids and erigesides.6Among them,only scutellarin,the most abundantflavone(accounted for0.56%of the dry herb10),attracted the attention of most investigators.In addition,many of its preparations including dispersible tablets,soft capsules,dripping pills,inter alia, have been or are being developed.11Preparations of herbal extracts,such as Dengzhanxixin injections and Dengzhan-xixin tablets,are also popular in China.12The upsurge in E.breviscapus related drug development has created a demand for fast,specific and sensitive ana-lytical methods.However,standard compounds,which are commercially unavailable in most cases,are usually necessary in most studies.As a result,many repetitive isolation and purification experiments,which are tedious, laborious and expensive,have had to be carried out. Moreover,some constituents in herbs are too low to enrich. The advent of high-performance liquid chromatography/ mass spectrometry(HPLC/MS)has thrown some light on the exploitation of complicated plant extracts,especially the minor components in them.13Recently,a variety of naturalRAPID COMMUNICATIONS IN MASS SPECTROMETRYRapid Commun.Mass Spectrom.2007;21:2971–2984Published online in Wiley InterScience()DOI:10.1002/rcm.3166*Correspondence to:H.Qu or Y.Cheng,College of Pharmaceutical Sciences,Zhejiang University(Zijingang Campus),Hangzhou 310058,P.R.China.E-mail:quhb@;chengyy@Contract/grant sponsor:National Basic Research Program(973) of the Ministry of Science and Technology of the P.R.China; contract/grant number:2005CB523402.Copyright#2007John Wiley&Sons,Ltd.products,such as flavonoids,phenolic acids,alkaloids and steroids,have been analyzed by HPLC/MS,which has displayed the power of HPLC/MS in structural character-ization of known or unknown constituents.14Qu et al .15identified nine flavonoids from the extract of E.breviscapus by a LC/MS/MS method.Yang et al .compared the main components in two kinds of preparations (tablets and injections)of E.breviscapus using HPLC coupled to diode array detection (DAD).16Although their clinical indications and usages are identical,very different component patterns were found for the two kinds of preparations.Scutellarin was the only main constituent in tablets while its concentration in injections was very low.The UV spectra of main components in injections were similar to those of caffeoyl derivatives but their identification was not obtained since no standards were available.So a comprehensive analysis of the phenolic compounds in E.breviscapus is in great demand.The aim of the present work was to develop a method based on HPLC/multistage mass spectrometry and DAD technologies for the identification of the phenolic com-ponents in E.breviscapus extract.After a solid-phase extrac-tion (SPE)step,separation was performed on a YMC C 18column.Negative ion mode was used in HPLC/MS and HPLC/MS n analysis.Finally,more than 50compounds were identified or tentatively characterized (Table 1).Most of them were reported for the first time.EXPERIMENTAL MaterialsHPLC-grade acetonitrile and methanol (Merck,Darmstadt,Germany)were used for HPLC analysis and sample extraction,respectively.Deionized water was purified using a Milli-Q system (Millipore,Bedford,MA,USA).Analytical-grade formic acid was purchased from Hangzhou Reagent Company (Hangzhou,China).Reference compounds (scu-tellarin,chlorogenic acid,caffeic acid,apigenin and luteolin)were purchased from the National Institute for the Control of Pharmaceutical and Biological Products (Beijing,China)and baicalin from Sigma (St.Louis,MO,USA).Erigoster A,erigoster B and a new compound,4-benzoyl-9-p -couma-royl-DOA,were separated from E.breviscapus and identified by nuclear magnetic resonance (NMR),UV and electrospray ionization multistage mass spectrometry (ESI-MS n )and by comparing with the reported data.17,18Their purities were above 95%,as determined by HPLC analysis.The whole plants of E.breviscapus were collected from the southern part of Yunnan Province.HLB Oasis 1SPE columns were purchased from Waters (Milford,MA,USA).Sample preparationThe whole plant of E.breviscapus was ground into fine powder (100mesh).A pulverized herbal sample (2g)was ultrasonically extracted with 50mL of 60%aqueous metha-nol for 30min.After centrifugation at 12000rpm (8791.2g )for 15min,the supernatant was transferred to a flask and evaporated in a rotatory evaporator (Buchi,Switzerland)until all the methanol had been removed.The residual aqueous solution was further purified by the following SPE method.An Oasis HLB SPE column was conditioned with 1mL methanol and equilibrated with 1mL acidic solution (1%formic acid in water).Then 0.5m L of E.breviscapus aqueous extract was loaded onto the column.After being washed with 0.5mL of the same acidic solution and 1mL methanol,respectively,the wash solution by methanol was collected for further analysis.An aliquot of 10m L was injected for HPLC/DAD and HPLC/MS analysis.HPLC systemLC analyses were performed on an Agilent 1100HPLC instrument (Agilent,Waldbronn,Germany)coupled to a binary pump,a diode-array detector (DAD),an autosampler,and a column compartment.The samples were separated on a YMC-Pack C 18column (4.6Â250mm,5m m).The mobile phase consisted of acetonitrile (A)and water containing 0.1%formic acid (B).The elution gradient was as follows:10%A (0min),17.5%A (20min),17.5%A (40min),45%A (80min)and 45%A (90min).The mobile phase flow rate was 0.8mL/min,and the column temperature was set at 308C.The on-line UV spectra were recorded in the range 190–400nm.Mass spectrometry and NMRFor LC/ESI-MS analyses,a Finnigan LCQ Deca XP plus ion trap mass spectrometer (Thermo Finnigan,San Jose,CA,USA)was connected to the Agilent 1100HPLC instrument via an ESI interface.The acquisition parameters were:collision gas,ultrahigh-purity helium (He);nebulizing gas,high purity nitrogen (N 2);ion spray voltage,À4.5kV;sheath gas (N 2)30arbitrary units;auxiliary gas (N 2)10arbitrary units;capillary temperature 3508C;capillary voltage À15V;tube lens offset voltage À30V;mass range recorded m/z 100–800.Data-dependent scans were performed to obtain the MS 2and MS 3spectra of all the components in the E.breviscapus extract.When MS n (n >3)scans were needed,the target peaks were collected after separation on the column and analyzed by direct loop injections.The MS n data of some low abundance compounds were also obtained through on-line two-stage full scan and multistage full scan.All the data are combined in Table 1and the specific methods used in our experiments are not indicated considering the conciseness of the table and the similarity of the spectra from different operating modes.A syringe pump was used for the direct loop injections of standard solutions (about 20m g/mL in methanol)and elutes of the main peaks.The flow rate was set at 10m L/min.Capillary temperature was set at 2758C and sheath gas 5arbitrary units.No auxiliary gas was needed.All other parameters were identical to those in LC/ESI-MS experiments.High-resolution mass spectrometry (HRMS)data was acquired on a hybrid QqTOF mass spectrometer (QSTAR XL,Applied Biosystems,Foster City,CA,USA)equipped with a TurboIonSpray source.The instrument was operated in the positive ion mode and full-scan spectra recorded in the range m/z 400–800.The NMR data were recorded on a 400MHz NMR instrument (Bruker Analytik GmbH)using standard pulse sequences.Copyright #2007John Wiley &Sons,Ltd.Rapid Commun.Mass Spectrom.2007;21:2971–2984DOI:10.1002/rcm2972Y.Zhang et al.Table 1.Chromatographic,UV and mass spectral characteristics of phenolic compounds from Erigeron breviscapusNo.t R (min)Identification [M–H]Àl max (nm)HPLC/(À)ESI-MS n m/z (%base peak)17.631-CQA 353–MS 2[353]:353(4),191(100),179(6)28.653-CQA 353236,300sh,324MS 2[353]:191(100),179(80),173(2),135(9)MS 3[353!191]:191(100),179(82),135(11)312.00mono-p -CoQA 337–MS 2[337]:191(5),173(2),163(100),119(3)412.605-CQA 353242,300sh,326MS 2[353]:191(100),179(5)MS 3[353!191]:191(100),173(14),127(8)513.354-CQA 353248,300sh,326MS 2[353]:191(18),179(99),173(100),155(2),135(10)MS 3[353!179]:179(100),135(14)6b13.809-COA397240,297sh,324MS 2[397]:397(5),353(23),335(9),293(100),281(21),235(65),221(26),203(9),191(6),179(48),161(30),135(6)MS 3[397!353]:353(37),335(61),307(3),281(13),273(5),263(5),245(12),221(4),219(2),203(2),179(100),135(5)MS 3[397!293]:293(31),275(100),249(10),233(17),216(6),215(7),203(43),179(15),165(4),161(42),135(6)7b 14.303-CDOA381240,299sh,324MS 2[381]:337(5),293(12),251(6),245(31),219(2),203(3),201(12),179(28),161(100),135(8),133(6),129(2)815.576,8-di-C -glucosyl apigenin 593–MS 2[593]:593(100),575(8),503(24),473(78),383(5),353(9)916.12Caffeic acid179240,300sh,324MS 2[179]:179(60),135(100)1016.30Malonyl-monoCQA439–MS 2[439]:395(100),353(3),233(16)MS 3[439!395]:335(3),233(100),173(7)MS 4[439!395!233]:233(2),173(100),155(6)11b 16.864-CDOA381240,300sh,324MS 2[381]:381(8),363(15),337(12),293(42),275(3),251(52),245(1),221(9),219(8),203(13),179(51),161(100),135(7)12b 17.371,3-diCQA glucoside 677–MS 2[677]:515(100),497(7),353(15),341(8),335(7)MS 3[677!515]:353(100),341(5),335(23),323(2),179(11)1318.14mono-p -CoQA 337236,300sh,318MS 2[337]:335(4),191(14),173(100),163(18)1419.011,3-diCQA515242,302sh,320MS 2[515]:353(100),335(30),191(3),179(20)MS 3[515!353]:191(100),179(75),135(8)15b 19.43Malonyl-monoCQA439246,300sh,324MS 2[439]:395(100),353(2),233(37)MS 3[439!395]:335(3),233(100),173(4)MS 4[439!395!233]:173(100),155(3),137(2)16b 19.589-CDOA381244,299sh,326MS 2[381]:337(7),293(25),251(34),245(5),219(2),203(4),201(2),179(20),161(100),135(6),133(6)17b24.523,9-di-CDOA glucoside705248,299sh,326MS 2[705]:543(100)MS 3[705!543]:543(38),407(2),381(100),363(7),319(13),283(4),221(11),203(4),179(8)MS 4[705!543!381]:363(10),337(6),295(2),293(25),259(2),251(43),245(6),221(4),219(3),203(7),179(37),161(100),135(5),133(3),129(3)1824.72Erigeroside 435–MS 2[435]:435(6),407(2),369(2),323(100),263(2),221(5),179(2),161(4)MS 3[435!323]:323(65),305(2),280(5),263(49),251(9),245(8),233(10),221(100),203(34),179(83),177(28),173(2),161(24),159(2),135(13)MS 4[435!323!221]:221(7),177(100)MS 5[435!323!221!177]:177(100),149(50)1927.143,4-diCQA glucoside 677238,300sh,322MS 2[677]:515(100),497(3),353(5)MS 3[677!515]:353(100),341(11),323(2),317(5),299(13),255(4),203(13),179(6),173(2)MS 4[677!515!353]:191(38),179(100),173(86),135(6)2028.36Carthamidin-7-O -glucuronide 463–MS 2[463]:301(10),287(100),269(3),175(5)MS 3[463!287]:287(10),269(13),259(24),243(21),193(13),181(100),167(58),153(27),139(5)2128.73Quercetin-3-O -glucuronide 477–MS 2[477]:397(2),373(6),371(2),343(3),301(100)MS 3[477!301]:301(99),273(15),257(11),255(3),239(6),229(3),179(100),169(3),151(57)2229.69Eriodictyol-7-O -glucuronide 463232,284,334MS 2[463]:463(2),301(13),300(4),287(100),175(11),151(7)MS 3[463!287]:287(3),269(3),151(100),135(3),125(2)2329.77Galetin-3-O -glucuronide477232,276,344MS 2[477]:301(100),233(13),191(4),149(2)MS 3[477!301]:301(100),283(6),255(6),245(6),229(4),211(2)2430.27Scutellarin 461232,282,334MS 2[461]:285(100)aMS 3[461!285]:285(100),267(39),241(8),239(22),213(10),119(3)(Continues )Copyright #2007John Wiley &Sons,Ltd.Rapid Commun.Mass Spectrom.2007;21:2971–2984DOI:10.1002/rcmPhenolic compounds in Erigeron breviscapus by LC/ESI-MS 2973Table 1.(Continued)No.t R (min)Identification[M–H]Àl max (nm)HPLC/(À)ESI-MS n m/z (%base peak)25b32.233,9-diCOA or 4,9-diCOA559246,298sh,330MS 2[559]:559(13),515(3),455(3),397(100),379(5),335(2),293(2),221(2),203(4),179(3)MS 3[559!397]:397(4),353(26),335(8),293(100),281(29),263(2),245(2),235(82),233(3),221(17),217(2),203(8),179(58),161(24),135(9)2632.931,4-diCQA 515246,292sh,330MS 2[515]:471(6),353(100),335(16),317(53),299(70),255(25),203(58),179(3),175(3)MS 3[515!353]:191(45),179(100),173(95),135(10)2734.163,4-diCQA 515242,300sh,324MS 2[515]:515(2),353(100),335(9),317(2),299(5),255(2),203(6),191(2),179(8),173(6)MS 3[515!353]:191(14),179(100),173(94)MS 4[515!353!179]:179(42),135(100)2834.78Kaempferol-3-O -rutinoside 593–MS 2[593]:593(39),285(100),284(3)aMS3[593!285]:285(20),257(14),255(100),227(13)2936.923,5-DiCQA515242,299sh,326MS 2[515]:353(100),335(1),191(3),179(1)MS 3[515!353]:353(10),191(81),179(100),173(5),135(7)MS 4[515!353!191]:191(100),127(25),85(7)3038.55Kaempferol-3-O -glucoside 447–MS 2[447]:447(100),327(5),285(39),284(21)aMS3[447!285]:285(12),257(18),255(100),227(15)3139.33Scutellarin-7-O -glucoside447–MS 2[447]:447(100),285(57),284(12)aMS 3[447!285]:285(100),284(23),283(10),267(38),241(10),239(16),228(6),213(7)32b40.891-Malonyl-3,5-diCQA 601242,301sh,328MS 2[601]:557(94),515(96),439(62),395(100),377(4),353(2),233(8)MS 3[601!557]:395(100),377(3),233(6)MS 3[601!515]:353(100),335(3),191(4)MS 3[601!395]:335(3),233(100),173(3)3344.00Narigenin-7-O -glucuronide 447–MS 2[447]:271(100),175(52),151(2)MS 3[447!271]:271(100),177(14),169(2),151(92)3444.94Erigoster B543240,299sh,326MS 2[543]:543(33),407(4),381(100),363(10),319(12),301(3),283(6),221(11),203(3),179(10)MS 3[543!381]:363(7),337(8),293(29),251(37),245(7),221(4),219(2),203(8),201(3),179(41),161(100),135(5),133(4),129(2)3546.934,5-diCQA 515234,300sh,326MS 2[515]:353(100),335(4),317(8),299(16),255(6),203(14),179(2),173(4)MS 3[515!353]:191(28),179(100),173(88),135(9)MS 4[515!353!179]:179(12),135(100)36b48.083,4-diCDOA or 4,9-diCDOA 543244,299sh,326MS 2[543]:543(28),499(3),455(2),381(100),363(2),319(14),251(5),221(4),203(3),179(8)MS 3[543!381]:363(11),337(15),293(34),251(85),221(9),219(11),203(11),201(2),179(51),161(100),135(7),129(7)3749.04Glucuronide of compound 42571336MS 2[571]:395(100)MS 3[571!395]:395(100),367(51),351(5),339(3),327(4),325(25),301(5),233(9)3849.18Syringic acid derivative 563232,280,334shMS 2[563]:383(43),365(47),321(2),197(100)MS 3[563!365]:197(100)MS 3[563!383]:197(4),185(100),141(2)MS 3[563!197]:197(63),182(19),153(100),138(9),121(3)MS 4[563!197!153]:153(5),138(100)39b49.494-Malonyl-3,5-diCQA 601–MS 2[601]:557(55),556(2),515(46),439(29),395(100),377(5),233(14)MS 3[601!557]:395(100),377(2),233(10)MS 3[601!515]:353(100),191(5),179(2)MS 3[601!395]:233(100),232(3)40b 50.46Acetyl-diCQA557–MS 2[557]:395(100),377(7),233(7)MS 3[557!395]:335(3),233(100),173(7)41b52.091-or 3-Malonyl-4,5-diCQA601240,300sh,328MS 2[601]:557(100),556(2),515(13),439(25),395(73)MS 3[601!557]:395(100)MS 3[601!515]:353(100),255(3)4252.82Unknown 395230,280,338MS 2[395]:395(100),367(50),351(5),339(3),327(4),325(24),301(5),233(10)4353.89Luteolin 285234,282,336aMS 2[285]:285(100),243(13),241(32),217(17),199(11),175(15)4454.421,3,5-triCQA677246,300sh,326MS 2[677]:515(32),497(100),469(2),353(14),335(7)(Continues )Copyright #2007John Wiley &Sons,Ltd.Rapid Commun.Mass Spectrom.2007;21:2971–2984DOI:10.1002/rcm2974Y.Zhang et al.RESULTS AND DISCUSSIONA solution of 60%methanol could extract more phenolic compounds than that of 100%methanol due to the high polarities of most components.Then the extracts were further enriched on a SPE column.Finally,10m L of each sample were injected for HPLC/DAD/MS analysis and the results are summarized in Table 1.In order to avoid confusion,the nomenclature of Yue et al .17for octulosonic acid derivatives has been used in this study (Fig.1).Representative chromatograms of E.breviscapus are presented in Fig. 2.As can be seen,the majority of compounds could be efficiently separated.Most constituents displayed a similar spectral behavior with two maximum absorption peaks at 230–240and 320–330nm and a shoulder at 290–300nm (Table 1).They were characterized as hydroxycinnamic acid derivatives.19Some peaks corre-sponding to flavones (three absorption peaks around 230,280and 330nm)were also observed.Due to the lack of reference compounds,HPLC/MS was utilized for furtherTable 1.(Continued)No.t R (min)Identification[M–H]Àl max (nm)HPLC/(À)ESI-MS n m/z (%base peak)MS 3[677!515]:353(100),335(4),179(4)MS 3[677!497]:497(19),335(100)MS 4[677!515!353]:191(100),179(82),135(5)4556.441,3,4-triCQA 677246,300sh,324MS 2[677]:515(15),497(100),317(6),299(10),255(2)MS 3[677!515]:515(8),472(2),353(100),341(6),339(2),335(2),299(3),255(3),203(2),173(3)MS 3[677!497]:335(11),317(44),299(100),273(4),255(42),203(7),179(14),161(3)MS 4[677!515!353]:191(12),179(100),173(90)46b57.653,4,9-triCOA 721244,299sh,320MS 2[721]:721(61),677(2),559(100),497(3),397(13)MS 3[721!559]:559(15),397(100),379(4),203(3)MS 4[721!559!397]:397(3),353(16),293(100),281(40),263(3),245(3),235(88),221(19),217(5),179(35),161(26)4758.29Eriodictyol287234,280,333sh aMS 2[287]:287(43),286(100),285(42),269(7),151(67)48b 59.844-Malonyl-1,3,5-triCQA 763244,300sh,320MS 2[763]:745(10),719(61),677(100),601(6),557(47),539(24),515(2),511(12),497(2),395(29),377(2)MS 3[763!677]:515(52),497(100),353(14)4961.03Scutellarein 285256,352aMS 2[285]:285(100),267(39),241(8),239(22),213(10),195(2),173(2),165(2),119(3)50b 62.31Acetyl-di-CQA 557–MS 2[557]:519(2),395(100)MS 3[557!395]:335(2),233(100),173(12)5162.973,4,5-TriCQA677246,300sh,326MS 2[677]:515(100),497(6)MS 3[677!515]:515(5),353(100),335(6),317(2),299(7),255(2),203(6),179(5),173(6)MS 4[677!515!353]:191(10),179(100),173(92)52b64.683,4,9-TriCDOA 705246,300sh,326MS 2[705]:705(70),661(12),543(100),481(19)MS 3[705!543]:543(40),381(100),363(7),319(18),283(6),179(8)MS 4[705!543!381]:363(17),337(5),295(4),293(22),251(34),233(12),221(10),219(9),215(8),203(20),179(25),161(100),135(5),123(13)5365.22Erigoster A 557–MS 2[557]:558(71),557(97),421(6),395(100),378(5),179(2),161(7)MS 3[557!395]:395(17),377(2),363(10),293(9),275(2),179(16),161(100),135(15),133(6)54b 66.313,4or 4,9-DiCDOA methyl ester 557–MS 2[557]:558(59),557(85),395(100),179(2),161(5)MS 3[557!395]:395(16),377(3),363(7),293(11),233(3),179(22),161(100),135(10),133(4)55b66.934-Benz-9-p -Co-DOA 469232,316MS 2[469]:425(13),347(100),323(5),303(6),275(5),235(20),201(2),187(8),185(7),183(12),163(62),145(61)MS 3[469!347]:303(100),275(9),235(23),213(7),201(4),187(7),185(16),183(7),163(91),145(37),119(10),107(4)MS 3[469!425]:425(7),303(100),213(7),189(7),145(7)MS 4[469!347!163]:163(22),119(100)MS 4[469!347!303]:303(41),243(8),199(7),163(28),145(100)5668.43Apigenin 269266,336aMS 2[269]:269(100),227(2),225(19),223(3),201(4),183(2),181(2),151(5),149(4)a Obtained with a collision energy of 40%.bNew compounds from Erigeron breviscapus .Copyright #2007John Wiley &Sons,Ltd.Rapid Commun.Mass Spectrom.2007;21:2971–2984DOI:10.1002/rcmPhenolic compounds in Erigeron breviscapus by LC/ESI-MS 2975Figure 1.The identified substructures from Erigeron breviscapus and ion nomenclature used forthem.Figure 2.HPLC/DAD/(–)ESI-MS analysis of the whole plant of Erigeron breviscapus :(A)DAD chromatogram monitored at 288nm and (B)total ion current chromatogram.Copyright #2007John Wiley &Sons,Ltd.Rapid Commun.Mass Spectrom.2007;21:2971–2984DOI:10.1002/rcm2976Y.Zhang et al.characterization of individual substances.During HPLC/MS analysis,a key step was to determine the molecular weight of each component ually,the most abundant peak in a full MS spectrum was assigned as [M–H]Àand,if adduct ions ([M–H þHCOOH]À)and dimers could be observed,this assignment was more solid.Furthermore,analyzing the same sample using acetic acid as an organic modifier (0.5%in water)provided more valuable information,such as [M–H þCH 3COOH]À,to support the identification of quasi-molecular ions in full MS spectra (data not shown).Identification of caffeoylquinic acids (CQAs)MonoCQAs (1,2,4,5)Compound 4was unambiguously identified as 5-CQA (chlorogenic acid)by comparison of retention time,UV and mass spectra with those of a reference substance.The occurrence of 5-CQA in natural plants or herbs is universal.20Furthermore,another three substances (compounds 1,2,5)yielded a [M–H]Àion at m/z 353and were identified as 1-CQA,3-CQA and 4-CQA (Table 1),respectively,whose fragmentation behavior was consistent with literature data.21,22The collision-induced dissociation (CID)spectrum of 1-CQA was identical with that of 5-CQA and they were discriminated by their elution order on reversed-phase (RP)-HPLC columns.The elution order of these four isomers was also in agreement with that reported by Clifford et al .22To our knowledge,the occurrence of 1-CQA,3-CQA and 4-CQA in E.breviscapus has not been reported previously.DiCQAs (14,26,27,29,35)In addition to the monocaffeoylquinic acids,several di-caffeoylquinic acid isomers were detected in E.breviscapus extracts (m/z 515,Table 1).These isomers were well studied in the ing the hierarchical key established by Clifford and co-workers,21,22they were identified as 1,3-diCQA (14),1,4-diCQA (26),3,4-diCQA (27),3,5-diCQA (29)and 4,5-diCQA (35)directly.TriCQAs (44,45,51)Compound 51expelled a caffeoyl moiety (162Da)to form a base peak at m/z 515([M–H–C]À)in MS/MS (Table 1).Its MS 3and MS 4spectra were identical with those of 3,4-diCQA.So compound 51was mostly 1,3,4-triCQA or 3,4,5-triCQA.In general,CQAs with a greater number of free equatorial hydroxyl groups in the quinic acid residue are more hydrophilic than those with a greater number of free axial hydroxyl groups.22Considering the long retention time of compound 51(t R ¼62.97min),it was plausibly identified as 3,4,5-triCQA.The caffeoyl moiety at C-5is easier to be lost than those at C-3and C-4,which is also in agreement with the reported data for CQAs.22Different from compound 51,a base peak at m/z 497([M–H–caffeic acid]À)was observed in CID spectra of compounds 44and 45,accompanied by a low intensity [M–H–C]Àion at m/z 515.Since we have little knowledge of the fragmentation behavior of [M–H–caffeic acid]Àions,we further analyzed the [M–H–C]Àions at m/z 515and compared with those of diCQAs.As a result,they were tentatively characterized as 1,3,5-triCQA and 1,3,4-triCQA,respectively.DiCQA glucosides (12,19)Another two compounds (12,19)showed the same [M–H]Àion at m/z 677as triCQAs but their retention times were much shorter.After a neutral loss of 162Da in MS 2,their MS 3showed the same characteristic ions with those of 1,3-diCQA and 3,4-diCQA,respectively.In natural pro-ducts,if not a caffeoyl moiety,the 162Da loss is always associated with a hexose (mostly a glucose).So compounds 12and 19were tentatively identified as 1,3-diCQA gluco-side and 3,4-diCQA glucoside,respectively.As we can see in the MS/MS spectra of compounds 12and 19,another ion at m/z 341([caffeic acid glucoside–H]À)was also observed,suggesting the glucose connected with one hydroxyl group of caffeic acid.23Malonyl-CQAs and acetyl-CQAs (10,15,32,39,40,41,48,50)Compounds containing malonyl or acetyl groups have not been reported from E.breviscapus up to now.However,these acyls are not uncommon in many plants.24–27Usually,they showed [M–H–44]Àand [M–H–86]Àions for a malonyl group and [M–H–60]Àfor a acetyl group in negative ESI-MS.28The identification of this type of compounds was illustrated by compound 32(Table 1).The MS/MS spectrum of compound 32showed characteristic ions of [M–H–44]Àat m/z 557and [M–H–86]Àat m/z 515,indicating a malonyl residue in its structure.Fragementation of the [M–H–86]Àion resulted in the identification of a 3,5-diCQA part.The malonyl group could not have been attached to a caffeoyl group because there was no loss of 206Da (acetyl-caffeoyl)but a loss of 162Da was observed in MS 3of [M–H–44]Àand [M–H–44–162]À.The [M–H–44–162–162]Àion at m/z 233was assigned as acetylquinic acid.We believed that such acetylation could stabilize the ring structure of quinic acid since ions corresponding to ring fragmentation were not observed.The malonyl group also could not condense with the C-1COOH to form an anhydride,which was not stable in an aqueous solution.So the malonyl group mostly con-densed with quinic acid at C-1or C-4to form an ester.Based only on the MS n data,we could not exclude any substituted form.Fortunately,based on the MS n data (Table 1),compound 39also possessed a malonyl residue and a 3,5-diCQA part in its structure.Similar to the rules for CQAs,21,22we could deduce that a 4-malonyl-QA has a longer retention time than a 1-malonyl-QA.Thus,compound 32was plausibly identified as 1-malonyl-3,5-diCQA and compound 39as 4-malonyl-3,5-diCQA.Based on this rule,compounds 41and 48were tentatively identified as 1-or 3-malonyl-4,5-diCQA and 4-malonyl-1,3,5-triCQA,pounds 10and 15were characterized as malonyl-monoCQAs but their [M–H–86]Àions were too low to identify the substitute position of caffeoyl groups.A 4-malonyl-3-CQA has been reported in Albizia julibrissin .29Just as we expected,the fragmentation behavior of [M–H–44]Àin MS 2of malonyl-CQAs was identical with that of the acetyl-CQAs (40,50).So the identification of acetyl-CQAs is straightforward.Two acetyl-CQAs (3-acetyl-4-CQA and 3-acetyl-5-CQA)have been found in eggplant (Solanum melongena L.).30Copyright #2007John Wiley &Sons,Ltd.Rapid Commun.Mass Spectrom.2007;21:2971–2984DOI:10.1002/rcmPhenolic compounds in Erigeron breviscapus by LC/ESI-MS 2977。
