Hydroxybenzoic and hydroxycinnamic acid derivatives in soft fruits

Quinic acid and glucose esters of hydroxycinnamic acids and glucosides of hydroxycinnamic and hydroxybenzoic acids have been determined by capillary GC and HPLC in soft fruits. The results confirm the wide distribution of these compounds in fruit tissues.     

苦丁茶中绿原酸及其异构体的提取变化分析

以富含绿原酸类成分的苦丁茶(Ilex kaushue)为材料,使用溶剂甲醇、乙醇、丙酮和水,结合超声波提取、水浴提取、回流提取等方法对绿原酸及其异构体的提取效率及提取后各异构体的变化进行分析。应用超高效液相色谱法可使苦丁茶的6种绿原酸类成分及咖啡酸在6 min内实现分离。提取结果表明,丙酮作为提取溶剂,在采用超声波和水浴提取时能获得较高的提取效率,易获得较多总量的绿原酸类成分和高含量异绿原酸A。而最常用的溶剂乙醇并未达到理想的提取效果。在不同溶剂的回流提取中,虽然提取的绿原酸类成分总量接近,但异绿原酸A和异绿原酸C含量有较大差异。醇溶液特别是乙醇溶液的回流提取使异绿原酸C的量大幅增加,而相应的异绿原酸A的量大幅减少,表明在醇加热条件下,异绿原酸A转化为异绿原酸C,这为获得抗氧化性更强的异绿原酸C提供了新思路。     

HPLC同时测定大叶冬青叶中10种多酚成分的含量及抗氧化活性研究

建立HPLC同时测定大叶冬青叶中新绿原酸、绿原酸、隐绿原酸、咖啡酸、对羟基肉桂酸、芦丁、异槲皮苷、异绿原酸B、异绿原酸A和异绿原酸C 10种多酚类化合物的定量分析方法,研究不同产地及不同采收时期大叶冬青叶中10种多酚类化合物的含量差异及其抗氧化活性。采用HPLC测定大叶冬青叶50%甲醇提取液中10种多酚类成分的含量,并选用DPPH法对其抗氧化活性进行初步探索。结果表明,10种多酚类化合物分离效果较好,标准曲线在检测范围内具有良好的线性(r>0.999 5),平均加样回收率在96.58%～112.03%,RSD<4%(n=6)。大叶冬青叶50%甲醇提取液抗氧活性良好。本实验建立的方法快速、准确、重复性好,可用于同时测定大叶冬青叶中新绿原酸、绿原酸、隐绿原酸、咖啡酸、对羟基肉桂酸、芦丁、异槲皮苷、异绿原酸B、异绿原酸A、异绿原酸C 10种成分的含量;不同产地、不同采收时期大叶冬青叶中10种多酚类化合物含量存在一定差异;不同来源大叶冬青叶50%甲醇提取液抗氧化能力具有明显差异。     

奎尼酸的生物合成与应用

奎尼酸是一种具有极高价值的精细化工产品和医药中间体,在医药、食品、化工等行业均有广泛的应用价值。奎尼酸广泛存在于多种植物中,其咖啡酰类衍生物在抗病毒、治疗心血管疾病等研究中得到越来越多的关注和应用。目前获得奎尼酸的方法主要有4种,即植物提取法、化学合成法、酶工程法和微生物发酵法。随着分子生物学的不断发展,结合现代生物技术,应用基因工程方法,必将成为获得奎尼酸的重要途径。本文综述奎尼酸的分布、生物合成及其应用。     

蔓三七叶中分离绿原酸和异绿原酸及其抗氧化活性研究

以蔓三七叶为原料,制备其中的绿原酸和异绿原酸A、B和C,并对它们进行结构鉴定;同时,采用DPPH法、水杨酸比色法、邻苯三酚自氧化法来对其进行体外抗氧化活性研究。结果表明,分离制备的绿原酸和异绿原酸A、B和C质量分数分别为96. 8%、98. 2%、97. 6%和98. 7%。在0. 5～10μg/mL范围内,绿原酸和异绿原酸A、B和C对DPPH和羟基自由基表现出了较好的清除作用,随浓度升高而逐渐增大;但它们对超氧自由基的清除作用时,浓度从5μg/mL升高到100μg/mL时,清除作用都未出现明显的剂量依赖性,各样品与阳性对照组(VC)相比较而言,其清除超氧自由基的作用明显较弱。本文揭示了蔓三七叶中的绿原酸和异绿原酸A、B和C具有不同的抗氧化效果。     

Intensity of radicle fluorescence as related to the resistance of seedlings of lettuce to the lettuce root aphid and carrot to the carrot fly

A technique is described which indicates by u.v. fluorescence the concentration of caffeoylquinic, chlorogenic and isochlorogenic acids in roots of germinating lettuce and carrot. The surface fluorescence of radicles from lettuce root aphid (Pemphigus bursarius) resistant seedlings was more intense than for susceptible seedlings, attributable to the higher concentration of isochlorogenic acid. In contrast, radicles of carrot seedlings resistant to carrot fly (Psila rosae) were less fluorescent than susceptible seedlings, corresponding to the lower concentration of chlorogenic acid in resistant seedlings. The technique is non-destructive and has been developed to distinguish between cultivars or breeding lines potentially resistant to these insect pests.     

不同甜叶菊品种叶中绿原酸类成分的比较研究

该文以14个扦插培育的甜叶菊品种叶为材料,从8种不同型号的树脂中筛选出一种合适的大孔吸附树脂对甜叶菊叶中绿原酸类成分进行纯化前处理,采用HPLC法对不同甜叶菊品种叶中所含绿原酸类成分进行比较分析。结果表明:(1)在8种不同型号的树脂中,XAD~(-1)6对甜叶菊叶中绿原酸类成分吸附-解析性能最佳。(2)经优化,上样液浓度1.20 mg·mL~(-1)、样品溶液pH 3、解析液乙醇体积分数70%时XAD~(-1)6树脂对甜叶菊叶中绿原酸类成分具有较好的纯化效果。(3) HPLC检测分析表明,在14个品种中共检测出新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸A、异绿原酸C六种绿原酸类成分,其中主要成分均为异绿原酸A、绿原酸、异绿原酸C,而在品种3、5、13、14中没有检测出异绿原酸B。(4) 14个品种中6个绿原酸类成分的含量分别为异绿原酸A 20.55~54.3 mg·g~(-1)、绿原酸17.96~32.93 mg·g~(-1)、异绿原酸C 4.15~19.49 mg·g~(-1)、新原酸0.61~4.61 mg·g~(-1)、隐绿原酸0.52~3.11 mg·g~(-1)、异绿原酸B 0.0~3.17 mg·g~(-1),6种绿原酸类成分总量为43.9~97.8 mg·g~(-1)。可见,不同品种甜叶菊叶中绿原酸类成分含量有明显差异,富含绿原酸类成分的甜叶菊品种可用于开发获取绿原酸类物质。     

Effect of Ethylene on the Phenylalanine Ammonia-lyase Activity of Carrot Tissues

Ethylene stimulated the activity of l -phenylalanine ammonia-lyase (PAL) in carrot (Daucus carota L.) tissues. It is also known to induce the formation of isochlorogenic acid. However, the induction patterns were different for isochlorogenic acid and PAL activity. Ethylene action seems to be indirect, as it did not affect the PAL activity of cell-free extracts.     

Phenolic acids associated with the resistance of lettuce cultivars to the lettuce root aphid

Phenolic acids, particularly caffeoylquinic acids, in lettuce root extracts were analysed by high pressure liquid chromatography (hplc), gas liquid chromatography (glc) and ultra-violet (uv) absorbance to seek a relationship with the resistance of certain lettuce cultivars to lettuce root aphid, Pemphigus bursarius. Although consistent results were obtained by each method, glc estimates of isochlorogenic acid tended to be low and uv estimates high. Hplc results were intermediate and since it was the easiest technique to perform routinely it was preferred. Isochlorogenic acid was the only caffeoylquinic acid detected in quantity and there were greater concentrations in resistant than in susceptible cultivars. Phenylalanine-ammonia lyase (PAL), the first enzyme in the phenyl propanoid pathway, is more active in resistant cultivars. Although these cultivars also had a greater tendency to browning when damaged, it was not due to greater polyphenol oxidase activity but probably to the presence of more isochlorogenic acid substrate. The results were consistent with an association between isochlorogenic acid concentration, PAL activity and resistance to P. bursarius.     

The Activity-Integrated Method for Quality Assessment of Reduning Injection by On-Line DPPH-CE-DAD

A sensitive on-line DPPH-CE-DAD method was developed and validated for both screening and determining the concentration of seven antioxidants of Reduning injection. The pH and concentrations of buffer solution, SDS, β-CD and organic modifier were studied for the detection of DPPH and seven antioxidants. By on-line mixing DPPH and sample solution, a DPPH-CE method for testing the antioxidant activity of the complex matrix was successfully established and used to screen the antioxidant components of Reduning injection. Then, antioxidant components including caffeic acid, isochlorogenic acid A, isochlorogenic acid B, isochlorogenic acid C, chlorogenic acid, neochlorogenic acid and cryptochlorogenic acid were quantified by the newly established CE–DAD method. Finally, the total antioxidant activity and the multiple active components were selected as markers to evaluate the quality of Reduning injection. The results demonstrated that the on-line DPPH-CE-DAD method was reagent-saving, rapid and feasible for on-line simultaneous determination of total pharmacological activity and contents of multi-components samples. It was also a powerful method for evaluating the quality control and mechanism of action of TCM injection.     

Evolution des Teneurs en Acides Chlorogénique et "Isochlorogénique' au Cours du Traitement de Vernalisation des Racines de Cichorium intybus

In a cold-requiring variety of endive (chicory, Cichorium intybus L. ev. Witloof) we could demonstrate a relationship between the variations in the content of chlorogenic acid during the cold treatment of the roots and their disposition to produce flowers in vitro. An increase in the level of chlorogenic acid in the roots preceded the development of their aptitude to form flower buds. In addition, the three main forms of “isochlorogenic’ acid are present in the roots of this cultivar: 3,5-dicaffeoyl-quinic acid, 3,4-dicaffeoyl-quinic acid, and traces of 4,5-dicaffeoyl-quinic acid. The variations in the levels of chlorogenic and “isochlorogenic’ acids indicate that part of the chlorogenic acid (3-caffeoyl-quinic acid) is converted to 3,5-dicaffeoyl-quinic acid.     

Studies on chlorogenic Acid biosynthesis in sweet potato root tissue in special reference to the isolation of a chlorogenic Acid intermediate

Marked polyphenol production takes place in root tissue of sweet potato, Ipomoea batatas Lam. cv. Norin 1, in response to slicing. A possible intermediate, tentatively termed compound V, of chlorogenic acid biosynthesis was isolated from the root tissue administrated with t-cinnamic acid-2-¹⁴C. Compound V was proved to be an ester whose acid moiety was t-cinnamic acid, and the hydroxyl group-bearing moiety appeared to be a carbohydrate. Compound V was suggested to be the first intermediate after t-cinnamic acid involved in the chlorogenic acid biosynthetic pathway by the following three results. (a) label of t-cinnamic acid-2-¹⁴C was distributed in compound V first, then transferred to chlorogenic acid and isochlorogenic acid, isomers of dicaffeoylquinic acid; (b) specific radioactivity of compound V increased prior to that of the fraction containing chlorogenic acid and isochlorogenic acids and decreased prior to that of the latter; and (c) label of compound V was efficiently incorporated into chlorogenic acid and isochlorogenic acid.     

Die organischen Säuren im Cambialsaft von Larix decidua Mill

Summary The quantitatively most important organic acids in the cambial sap of Larix decidua have been isolated and characterised. Quinic acid predominated and smaller amounts of shikimic acid, malic acid, oxalic acid, amino acids and sugar phosphates were also present. It is shown by the aphid-technique that quinic acid is probably transported in the phloem from the young needles and shoots to the cambial zones of the tree.

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Zweite Mitteilung über den Cambialsaft von Larix decidua. Erste Mitteilung in Planta (Berl.) 73, 155–160 (1967).     

忍冬属植物开花过程中绿原酸类化合物含量的变化

忍冬是中国传统中药材金银花的药源植物。采用高效液相色谱—光电二极管阵列检测器联用技术(HPLC-PAD)和高效液相色谱—电喷雾离子化—多级质谱联用技术(HPLC-ESI-MSn)对忍冬和4种同属植物在开花不同阶段中绿原酸类化合物进行了定性和定量分析,以明确忍冬属植物花的利用价值及其最佳采收期。结果发现:4种忍冬属植物花蕾及开放花朵中均含有新绿原酸、绿原酸、绿原酸甲酯、异绿原酸A和一种异绿原酸的异构体;绿原酸和总绿原酸在花蕾中的含量高于开放花朵,但贯月忍冬变化不显著;火焰忍冬、贯月忍冬、台尔曼忍冬中绿原酸和总绿原酸的含量在花蕾膨大期(大白期)和初花期(银花期)都高于忍冬,而格雷姆忍冬和忍冬差异不显著,研究表明,4种忍冬属植物花及花蕾都具有较高的开发利用价值。     

Turnover and metabolism of chlorogenic Acid in xanthium leaves and potato tubers

The active turnover of chlorogenic acid (3-caffeoylquinic acid(3)), a major phenolic component of Xanthium leaves and potato tuber disks, has been demonstrated in these tissues. Pulse-labelling experiments with radioactive l-phenylalanine and trans-cinnamic acid as well as direct feeding experiments with chlorogenic acid-(14)C labelled in the caffeoyl moiety have been employed in the turnover studies. The rate of turnover is calculated to be on the order of 50 to 100 mmumoles per hour per gram fresh weight of tissue.In Xanthium leaves chlorogenic acid is in part converted to an isochlorogenic acid identified by silica gel chromatography as 3,5-dicaffeoylquinic acid. Radioactivity of the caffeoyl moiety of chlorogenic acid is also incorporated into lignin-like insoluble polymers in the leaf. Turnover of chlorogenic acid in tuber tissue is largely accounted for by the incorporation of the caffeoyl moiety into insoluble polymers in the tissue.The significance of chlorogenic acid turnover is discussed in relation to the perception of the photoperiodic stimulus by leaves and to the possible role of chlorogenic acid in lignin synthesis.     

Effect of Daylength on Phenol Metabolism in the Leaves of Salvia occidentalis

A method of phenol determination in plant leaves has been developed which is based on the in situ oxidation of these compounds in an atmosphere containing ammonia, followed by difference spectrophotometry. The development of the phenol pattern has been studied in each separate leaf of a Salvia occidentalis plant grown in short and in long days. During the light period the phenol content (mainly chlorogenic acid and isochlorogenic acids) increases in proportion to the length of this period, whereas during the subsequent dark period the phenol content decreases. This decrease does not continue during the second part of a dark period if that period is interrupted by a light break with red light. Instead a small increase is observed. This effect of red light can be reversed with far red light. It is argued that a correlation with flower induction in this short day plant can be construed if it is assumed that the continuous presence of certain o-dihydroxyphenols in the cytoplasm of leaf cells inhibits the synthesis or the transport of a flowering hormone.     

Studies on auxin protectors XIV. Chlorogenic acid, a low molecular weight auxin protector in sunflower

Abstract Acetone extracts of sunflower leaves yielded two low molecular weight compounds with significant auxin protector activity. By means of chromatography, UV spectra, and acid hydrolysis it was possible to ascertain that most of the protector activity was associated with chlorogenic acid. Some protector activity was also associated with a derivative of chlorogenic acid, tentatively identified as isochlorogenic acid. A possible role for chlorogenic acid is proposed.     

Relations polyphénols — croissance: Mise en évidence d'un effet inhibiteur des composés phénoliques sur le transport polarisé de l'auxine

Polyphenols and Growth: Inhibition of Polar Auxin Transport by Phenolic Compounds. The possible effects of polyphenols on auxin transport in tomato plants (Lycopersicum esculentum Mill.) were investigated. For this purpose, the phenolic content of the material was stimulated by exogenously supplied quinic acid. After the apical bud had been excised, labelled compounds were applied to the cut surface, and the radioactivity transported to the roots was measured. Quinic acid treatment significantly delayed polar transport of labelled auxins (IAA or NAA). It did not affect the migration rate of sucrose^?14C and leucine^?3H. A number of evidences seems to demonstrate that the phenolics are responsible for these modifications, since similar results were recorded when the labelled compounds were supplied simultaneously with polyphenols from tomato. Moreover, a decreased polarity of NAA transport could be observed when the plants were submitted to treatments which lead to an increased level of phenols (boron deficiency, infection by Fusarium oxysporum). The data presented in this paper suggest that phenolic compounds could act on growth processes via the regulation of polar auxin transport.     

Changes in bulk protein of tobacco leaves on aerobic autolysis: Hydrogenation studies and identification of bound quinic acid

During aerobic autolysis and in commercial curing, the bulk proteins of tobacco leaves become coupled with quinic acid, presumably in consequence of coupling of chlorogenic acid congeners with lysine ε-NH₂ groups. Quinic acid derivatives, prepared from acid hydrolysates of such altered proteins, were identified by GC-MS. Such proteins were also hydrogenated over Rh/Al₂O₃ with a view to stabilizing the hypothetical linkages. Difficulties in removing contaminant Al had to be overcome. Evidence was then obtained (by GLC of derivatives) for several components, in acid hydrolysates of hydrogenated altered proteins, which were neither normal hydrogenation products of the common amino acids nor derivatives of quinic acid. Details of the chromatograms and mass spectra of quinic acid derivatives are provided in a supplementary publications.     

Flavonoids from Achyrocline flaccida

Three new flavonoids: 5-hydroxy-7-(3-methyl-2,3-epoxybutoxy)flavanone,5-hydroxy-3,8-dimethoxy 7-(3-methyl-2,3-epoxybutoxy)flavone and 4′-hydroxy-5-methoxy-7-(3-methyl-2,3-epoxybutoxy)flavone were isolated and identified from the aerial parts of Achyrocline flaccida. Tamarixetin, gnaphaliin, isognaphaliin, 5,7,8-trihydroxy-3-methoxyflavone, chrysoeriol, galangin 3-methyl ether, naringenin 5-methyl ether, caffeic acid, chlorogenic acid and isochlorogenic acid were also isolated.