Molecular definition of the taste of roasted cocoa nibs (Theobroma cacao) by means of quantitative studies and sensory experiments

Sensory-guided decomposition of roasted cocoa nibs revealed that, besides theobromine and caffeine, a series of bitter-tasting 2,5-diketopiperazines and flavan-3-ols were the key inducers of the bitter taste as well as the astringent mouthfeel imparted upon consumption of roasted cocoa. In addition, a number of polyphenol glycopyranosides as well as a series of N-phenylpropenoyl-l-amino acids have been identified as key astringent compounds of roasted cocoa. In the present investigation, a total of 84 putative taste compounds were quantified in roasted cocoa beans and then rated for the taste contribution on the basis of dose-over-threshold (DoT) factors to bridge the gap between pure structural chemistry and human taste perception. To verify these quantitative results, an aqueous taste reconstitute was prepared by blending aqueous solutions of the individual taste compounds in their "natural" concentrations. Sensory analyses revealed that the taste profile of this artificial cocktail was very close to the taste profile of an aqueous suspension of roasted cocoa nibs. To further narrow down the number of key taste compounds, finally, taste omission experiments and human dose/response functions were performed, demonstrating that the bitter-tasting alkaloids theobromine and caffeine, seven bitter-tasting diketopiperazines, seven bitter- and astringent-tasting flavan-3-ols, six puckering astringent N-phenylpropenoyl-l-amino acids, four velvety astringent flavonol glycosides, gamma-aminobutyric acid, beta-aminoisobutyric acid, and six organic acids are the key organoleptics of the roasted cocoa nibs.     

Identification of the astringent taste compounds in black tea infusions by combining instrumental analysis and human bioresponse

Application of taste dilution analyses on freshly prepared black tea infusions revealed neither the high molecular weight thearubigen-like polyphenols nor the catechins and theaflavins, but a series of 14 flavon-3-ol glycosides as the main contributors to the astringent taste perceived upon black tea consumption. Among these glycosides, the apigenin-8-C-[alpha-l-rhamnopyranosyl-(1-->2)-O-beta-d-glucopyranoside] was identified for the first time in tea infusions. Depending on the structure, the flavon-3-ol glycosides were found to induce a velvety and mouth-coating sensation at very low threshold concentrations, which were far below those of catechins or theaflavins; for example, the threshold of 0.001 micromol/L found for quercetin-3-O-[alpha-l-rhamnopyranosyl-(1-->6)-O-beta-d-glucopyranoside] is 190000, or 16000 times below the threshold determined for epigallocatechin gallate or theaflavin, respectively. Moreover, structure/activity considerations revealed that, besides the type of flavon-3-ol aglycon, the type and the sequence of the individual monosaccharides in the glycosidic chain are key drivers for astringency perception of flavon-3-ol glycosides.     

Identification and comparison of phenolic compounds in the preparation of oolong tea manufactured by semifermentation and drying processes

Oolong tea manufactured via a semifermentation process possesses a taste and color somewhere between green and black teas. Alteration of constituents, particularly phenolic compounds, in the infusion of oolong tea resulting from its manufacture, was analyzed by high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry. The identified constituents contained 2 alkaloids, 11 flavan-3-ols, 8 organic acids and esters, 11 proanthocyanidin dimers, 3 theaflavins, and 22 flavonoid glycosides, including 6 novel acylated flavonol glycosides. The tentative structures of these 6 novel compounds were depicted according to their mass fragmentation patterns in MS(n) (n = 1-4). In comparison with caffeine as an internal standard, relative contents of the constituents in the infusions of fresh tea shoot and different oolong tea preparations were examined. Approximately, 30% catechins and 20% proanthocyanidins were oxidized during the manufacture of oolong tea from fresh tea shoots, and 20% of total flavonoids were decomposed in a follow-up drying process. Gallocatechin-3-O-gallate and theaflavins putatively produced in the semifermentation process of oolong tea were not detected in fresh tea shoots, and the majority of theaflavins were presumably transformed into thearubigins after drying.     

Orosensory-directed identification of astringent mouthfeel and bitter-tasting compounds in red wine

Application of sequential solvent extraction, followed by HPLC combined with the taste dilution analysis, enabled the localization of the most intense velvety astringent, drying, and puckering astringent, as well as bitter-tasting, compounds in red wine, respectively. Isolation of the taste components involving gel adsorption chromatography, ultrafiltration, and synthesis revealed the identification of 26 sensory-active nonvolatiles, among which several hydroxybenzoic acids, hydroxycinnamic acids, flavon-3-ol glycosides, and dihydroflavon-3-ol rhamnosides as well as a structurally undefined polymeric fraction (>5 kDa) were identified as the key astringent components. In contradiction to literature suggestions, flavan-3-ols were found to be not of major importance for astringency and bitter taste, respectively. Surprisingly, a series of hydroxybenzoic acid ethyl esters and hydroxycinnamic acid ethyl esters were identified as bitter compounds in wine. Taste qualities and taste threshold concentrations of the individual wine components were determined by means of a three-alternative forced-choice test and the half-mouth test, respectively.     

Quantitative studies, taste reconstitution, and omission experiments on the key taste compounds in morel mushrooms (Morchella deliciosa Fr.)

Sensory-directed fractionation of an aqueous extract prepared from morel mushrooms led to the identification of gamma-aminobutyric acid as the chemical inducer of the mouth-drying and mouth-coating oral sensation imparted by morels. Additionally, L-glutamic acid, L-aspartic acid, succinic acid, and the previously unknown (S)-malic acid 1-O-beta-D-glucopyranoside, coined (S)-morelid, were detected as additional important umami-like taste compounds. To further bridge the gap between pure structural chemistry and human taste perception, 33 putative taste compounds were quantified in an aqueous morel extract and then rated for their taste contribution on the basis of dose-over-threshold factors. To confirm these quantitative results, an aqueous taste reconstitute was prepared by blending aqueous solutions of 16 amino acids, 6 organic acids, 3 purines, 4 carbohydrates, 3 minerals, and (S)-morelid in their "natural" concentrations. Triangle tests revealed that the taste profile of this biomimetic organoleptic cocktail did not differ significantly from the taste profile of authentic morel extract. To finally narrow down the number of key taste compounds, taste omission experiments were performed demonstrating that (S)-morelid together with L-glutamic acid, L-aspartic acid, malic acid, citric acid, acetic acid, and gamma-aminobutyric acid are the key organoleptics of morel extract. Moreover, sensory experiments with model solutions showed that (S)-morelid not only imparts a sour and umami-like taste but is able to amplify the taste activity of monosodium glutamate, as well as sodium chloride, solutions.     

Characterization of an intense bitter-tasting 1H,4H-quinolizinium-7-olate by application of the taste dilution analysis, a novel bioassay for the screening and identification of taste-active compounds in foods

Thermal treatment of aqueous solutions of xylose and primary amino acids led to rapid development of a bitter taste of the reaction mixture. To characterize the key compound causing this bitter taste, a novel bioassay, which is based on the determination of the taste threshold of reaction products in serial dilutions of HPLC fractions, was developed to select the most intense taste compounds in the complex mixture of Maillard reaction products. By application of this so-called taste dilution analysis (TDA) 21 fractions were obtained, among which 1 fraction was evaluated with by far the highest taste impact. Carefully planned LC-MS as well as 1D and 2D NMR experiments were, therefore, focused on the compound contributing the most to the intense bitter taste of the Maillard mixture and led to its unequivocal identification as the previously unknown 3-(2-furyl)-8-[(2-furyl)methyl]-4-hydroxymethyl-1-oxo-1H,4H-quinolizinium-7-olate. This novel compound, which we name quinizolate, exhibited an intense bitter taste at an extraordinarily low detection threshold of 0.00025 mmol/kg of water. As this novel taste compound was found to have 2000- and 28-fold lower threshold concentrations than the standard bitter compounds caffeine and quinine hydrochloride, respectively, quinizolate might be one of the most intense bitter compounds reported so far.     

Reconstitution of the flavor signature of Dornfelder red wine on the basis of the natural concentrations of its key aroma and taste compounds

By application of aroma extract dilution analysis (AEDA) on the volatile fraction isolated from a Dornfelder red wine, 31 odor-active compounds were identified by means of HRGC-MS and comparison with reference compounds. A total of 27 odorants, judged with high FD factors by means of AEDA, was quantitated by means of stable isotope dilution assays, and acetaldehyde was determined enzymatically. In addition, 36 taste-active compounds were analyzed by means of HPLC-UV, HPLC-MS/MS, and ion chromatography. The quantitative data obtained for the identified aroma and taste compounds enabled for the first time the reconstruction of the overall flavor of the red wine. Sensory evaluation of both the aroma and taste profiles of the authentic red wine and the recombinate revealed that Dornfelder red wine was closely mimicked. Moreover, it was demonstrated that the high molecular weight fraction of red wine is essential for its astringent taste impression. By comparison of the overall odor of the aroma recombinate in ethanol with that of the total flavor recombinate containing all tastants, it was shown for the first time that the nonvolatile tastants had a strong influence on the intensity of certain aroma qualities.     

Factors affecting the caffeine and polyphenol contents of black and green tea infusions.

The effects of product and preparation variables on the in-cup chemical composition of tea extracts is of interest because the appearance and taste characteristics and the possible health effects of a tea liquor arise from the chemical components extracted from the leaf during tea preparation. A comprehensive study was therefore undertaken to determine the contributions of product and preparation variables on the total soluble solids, caffeine, and polyphenol contents of tea extracts. The results of this study show that the variety, growing environment, manufacturing conditions, and grade (particle size) of the tea leaves each influence the tea leaf and final infusion compositions. In addition, the composition of the tea infusion was shown to be influenced by whether the tea was contained in a teabag and, if so, the size and material of construction of the bag. Finally, the preparation method, including the amounts of tea and water used, infusion time, and amount of agitation, was shown to be a major determinant of the component concentrations of tea beverages as consumed. An illustration of the variation introduced by these product and preparation factors is provided by comparing solids, caffeine, and polyphenol contents of green and black tea infusions when commercial products are prepared according to the instructions given on their packaging.     

不同茶树品种(系)的绿茶滋味分析及评价模型构建

为客观准确地评价不同茶树品种(系)绿茶的滋味品质,该研究系统分析了25个茶树品种(系)绿茶的主要滋味成分含量及其Dot值,利用主成分分析法对不同茶树品种(系)绿茶滋味品质进行综合评价并建立滋味品质评价模型。结果表明:儿茶素、咖啡碱是不同茶树品种(系)绿茶苦涩味强度差异的主要原因,其中表没食子儿茶素没食子酸酯(Epigallocatechingallate,EGCG)是所测样品中涩味的主要贡献物质,EGCG和咖啡碱为所测样品中苦味的主要贡献物质。氨基酸是不同茶树品种(系)绿茶鲜味、甜味差异主要因素,谷氨酸是所测样品中鲜味的主要贡献物质。主成分分析表明:前5个主成分的累计方差贡献率为79.018%,并以前5个主成分的线性回归方程和贡献率构建了滋味品质评价模型,模型评价结果与感官审评结果较为相似,存在极显著相关性(P0.01),相关系数为0.791;利用模型中各主成分的得分,能够较好的区分所测样品的苦、涩、鲜、爽等滋味特征。因此,该研究所建模型能够较好的评价所测样品的滋味品质及其主要滋味特征的差异,为各茶树品种(系)的推广应用提供理论依据,也为不同茶树品种(系)绿茶滋味品质的科学评价提供新的思路和方法。     

HPLC-MSn analysis of phenolic compounds and purine alkaloids in green and black tea

Tea is a complex mixture containing a range of compounds from simple phenolics to complex thearubigins, many of which have well-recognized antioxidant properties. This paper describes the application of high-performance liquid chromatography-mass spectrometry (HPLC-MS(n)) methods for the rapid and routine analysis of more than 30 phenolics in tea. Green and black tea infusions were injected directly onto a reversed phase HPLC column, and the phenolics eluted using two different mobile phase gradients, one optimized to resolve catechin derivatives and the other, flavonols and theaflavins. Compounds, identified on the basis of their retention time, absorbance spectrum, and MS fragmentation pattern, included (+)-catechin, (-)-epicatechin, theaflavin and their various gallate derivatives, quercetin and kaempferol mono-, di-, and triglycosides, quinic acid esters of gallic acid and hydroxycinnamates, and the purine alkaloids, caffeine and theobromine.     

采用PLS-DA分析毛火方式对工夫红茶品质的影响

为探究不同毛火方式对工夫红茶品质的影响,明确新型电磁内热式滚筒-热风耦合干燥设备的毛火效果,该研究以一芽一二叶初展嫩度的"福鼎大白"品种为原料进行工夫红茶加工,设定电磁滚筒-热风耦合(Rotary pot-Hot air coupling First-Drying with electromagnetic heat,RHFD)、链板热风(Chain plate Hot air First-Drying,CHFD)、箱式热风(Box Hot air First-Drying,BHFD)、滚筒式滚炒(Rotary pot First-Drying,RFD)等4种毛火方式,比较所制茶样的茶多酚、儿茶素、茶色素、可溶性糖、咖啡碱、氨基酸等29个非挥发性指标,114个气相色谱-质谱技术(GasChromatography-Mass Spectrometry,GC-MS)检测的挥发性香气指标,10个外形和汤色色泽客观评价指标,同时进行了毛火方式的热效率、生产效率、生产成本等性能指标的分析比较,通过偏最小二乘判别统计(PartialLeastSquaresDiscriminationAnalysis,PLS-DA)分析毛火方式对优质工夫红茶品质的影响,并获得标志性差异化合物。结果表明:电磁内热式滚筒-热风耦合毛火处理下茶多酚和儿茶素总量显著最低(P<0.05),简单儿茶素含量较高,茶红素和可溶性糖含量、茶黄素综合指标TDE和茶色素综合指标10TFRB最高(P<0.05),毛火方式对茶黄素总量影响不显著(P>0.05);挥发性化合物总量以RHFD方式最高,RFD方式次之,CHFD方式最低;RHFD毛火方式芳香类、萜烯类等化合物含量最高。电磁内热式滚筒-热风耦合毛火升温快、温度分布均匀且稳定性好,热效率和生产效率高(分别为50.0%、220 kg/h),生产成本较低(仅0.32元/kg),预热时间仅14min;所制红茶在汤色透亮度、香气甜久度、滋味甜醇度等方面均得到提升,感官总分最高(P<0.05),达88.1。PLS-DA分析从挥发性和非挥发性角度均可将工夫红茶4种毛火方式显著区分,并分别获得了43种和18种差异化合物,结合差异性分析获得标志性差异化合物,2,4,6-三(1,1-二甲基乙基)-4-甲基环己-2,5-二烯-1-酮、香叶醇、3-辛酮、水杨酸甲酯、茶黄素、茶褐素、可溶性糖、表儿茶素等,可作为区分工夫红茶毛火方式,以及定向加工甜香、甜醇、高亮等优质工夫红茶的指标物质。该研究为红茶加工基础和品质提升提供技术参考和理论指导。     

Characterization of the key aroma compounds in the beverage prepared from Darjeeling black tea: quantitative differences between tea leaves and infusion

By application of the aroma extract dilution analysis on the volatile fraction isolated from a black tea infusion (Darjeeling Gold Selection), vanillin (vanilla-like), 4-hydroxy-2,5-dimethyl-3(2H)-furanone (caramel), 2-phenylethanol (flowery), and (E,E,Z)-2,4,6-nonatrienal (oat-flake-like) were identified with the highest flavor dilution (FD) factors among the 24 odor-active compounds detected in the FD factor range of 4-128. Quantitative measurements performed by means of stable isotope dilution assays and a calculation of odor activity values (OAVs; ratio of concentration to odor threshold in water) revealed, in particular, the previously unknown tea constituent (E,E,Z)-2,4,6-nonatrienal as a key odorant in the infusion and confirmed the important role of linalool and geraniol for the tea aroma. An aroma recombinate performed by the 18 odorants for which OAVs > 1 were determined in their "natural" concentrations matched the overall aroma of the tea beverage. In the black tea leaves, a total of 42 odorants were identified, most of which were identical with those in the beverage prepared thereof. However, quantitative measurements indicated that, in particular, geraniol, but also eight further odorants were significantly increased in the infusion as compared to their concentration in the leaves.     

Identification of the taste enhancer alapyridaine in beef broth and evaluation of its sensory impact by taste reconstitution experiments

An essential compound imparting the sweet taste to beef broth was investigated. Taste activity-guided fractionation of beef broth by ultrafiltration, gel permeation chromatography, and HPLC in combination with the recently developed comparative taste dilution analysis enabled the localization of a fraction possessing sweetness-enhancing activity upon degustation. Comparison of the chromatographic, spectroscopic, and sensory data with those of the synthetic reference compound led to the identification of the sweetness-enhancing N-(1-carboxyethyl)-6-(hydroxymethyl)pyridinium-3-ol inner salt, named alapyridaine, which was recently isolated from heated aqueous solutions of hexoses and l-alanine. After quantification of alapyridaine in beef broth, sensory analysis of synthetic beef taste recombinates spiked with synthetic alapyridaine in its "natural" concentration of 419 mug/L and comparison to the taste quality of a tastant recombinate lacking the alapyridaine revealed a significant increase in sweetness and umami character only when the alapyridaine was present in the recombinate. These data demonstrate for the first time that, in "natural" concentrations, the alapyridaine exhibited a pronounced effect on the overall taste quality of beef broth, in particular, on the sweet and umami character.     

Hydration properties and the role of water in taste modalities of sucrose, caffeine, and sucrose-caffeine mixtures

Solution properties of sapid molecules are informative on their type of hydration (hydrophobic or hydrophilic) and on the extent of the hydration layer. Physicochemical properties (intrinsic viscosity and apparent specific volume) and nuclear magnetic resonance (NMR) relaxation rates R(1) and R(2) for pure sucrose, bitter molecule caffeine, and their mixture were found to be relevant in the interpretation of the effects of these solutes on water mobility. Likewise, surface tension, contact angles with a hydrophobic surface, and the adhesion forces to this type of surface of the aqueous solutions of sapid molecules were found to discriminate between their effects on water cohesion and also between their taste qualities. The interpretation of the two sets of independent experimental results, namely physicochemical and spectroscopic data, helps in the elucidation of the role of water in sweet and bitter taste chemoreception.     

Sensory-guided decomposition of roasted cocoa nibs (Theobroma cacao) and structure determination of taste-active polyphenols

Sequential application of solvent extraction, gel permeation chromatography, and RP-HPLC in combination with taste dilution analyses, followed by LC-MS and 1D/2D-NMR experiments and thiolytic degradation, revealed that, besides theobromine and caffeine, the flavan-3-ols epicatechin, catechin, procyanidin B-2, procyanidin B-5, procyanidin C-1, [epicatechin-(4beta-->8)](3)-epicatechin, and [epicatechin-(4beta-->8)](4)-epicatechin were among the key compounds contributing to the bitter taste as well as the astringent mouthfeel imparted upon consumption of roasted cocoa. In addition, a series of quercetin, naringenin, luteolin, and apigenin glycopyranosides as well as a family of not previously identified amino acid amides, namely, (+)-N-[4'-hydroxy-(E)-cinnamoyl]-L-aspartic acid, (+)-N-[3',4'-dihydroxy-(E)-cinnamoyl]-L-aspartic acid, (-)-N-[3',4'-dihydroxy-(E)-cinnamoyl]-L-glutamic acid, (-)-N-[4'-hydroxy-(E)-cinnamoyl]-L-glutamic acid, (-)-N-[4'-hydroxy-(E)-cinnamoyl]-3-hydroxy-L-tyrosine, (+)-N-[4'-hydroxy-3'-methoxy-(E)-cinnamoyl]-L-aspartic acid, and (+)-N-(E)-cinnamoyl-L-aspartic acid, have been identified as key astringent compounds of roasted cocoa. Furthermore, (-)-N-[3',4'-dihydroxy-(E)-cinnamoyl]-3-hydroxy-l-tyrosine (clovamide), (-)-N-[4'-hydroxy-(E)-cinnamoyl]-L-tyrosine (deoxyclovamide), and (-)-N-[3',4'-dihydroxy-(E)-cinnamoyl]-L-tyrosine, reported previously as antioxidants, have been found as contributors of cocoa's astringent taste. By means of the half-tongue test, the taste thresholds of flavan-3-ols and glycosides have been determined.     

New phenolic components and chromatographic profiles of green and fermented teas

A standardized profiling method based on liquid chromatography with diode array and electrospray ionization mass spectrometric detection (LC-DAD-ESI/MS) was applied to establish the phenolic profiles of 41 green teas and 25 fermented teas. More than 96 phenolic compounds were identified that allowed the teas to be organized into five groups. Epigallocatechin gallate (EGCG) was the major phenolic component of green tea made from mature leaves (group 2), while green tea made from the younger buds and leaves (group 1) contained lower flavonoid concentrations. Partially fermented teas (group 3) contained one-half the EGCG content of the green tea. Fully fermented black teas (group 4) had a trace of EGCG, but contained theaflavins. Highly overfermented black tea (group 5) contained only trace amounts of flavonol glycosides and theaflavins. Over 30 phenolics are new for tea, and this is the first phenolic profile to simultaneously detect C- and O-glycosylated flavonoids, catechins, proanthocyanidins, phenolic acid derivatives, and purine alkaloids.     

Determination of theaflavins including methylated theaflavins in black tea leaves by solid-phase extraction and HPLC analysis

A quantitative method for four theaflavins and two methylated theaflavin derivatives in black tea leaves was developed by solid-phase extraction and a high-performance liquid chromatographic method with photodiode array detection. The theaflavins in black tea leaves were extracted three times with 40 vol 50% aqueous ethanol (mg dry tea powder/mL) containing 2% ascorbic acid. The ethanol extracts were diluted 4-fold with distilled water. All diluted extracts were directly applied to the solid-phase C18 cartridge column without concentration. The fraction of theaflavins was obtained by 40% ethanol extraction after rinsing with water followed with 15% ethanol extraction. An aliquot of theaflavins after concentration was injected onto an ODS C18 reversed-phase column, and four theaflavins and two methylated theaflavins were sufficiently separated by a linear gradient system using distilled water and acetonitrile with 0.5% acetic acid. This analytical method is sensitive for the determination of a small amount of methylated theaflavins, since various interfering substances produced during the fermentation process were eliminated in advance by solid-phase extraction. Using this analytical method, we also demonstrated that methylated theaflavins were easily produced during the manufacture of black tea.     

不同防冻措施茶园茶汤滋味差异的电子舌检测

为探讨轻度“倒春寒”对不同防护措施下3种茶园蒸青茶样的茶汤滋味影响,遂选品种和管理措施一致的安装防霜扇的茶园、松树林围绕的茶园以及未设防护措施的普通茶园,用电子舌综合评价3种茶园茶汤滋味、高效液相色谱检测茶汤游离氨基酸组分及含量。结果表明：基于电子舌检测数据的主成分分析、软独立建模分析和聚类分析结果揭示了普通茶园的茶汤滋味与防霜扇茶园和林间茶园的差异显著（P<0.05）;从9种茶样中皆检测出18种氨基酸,游离氨基酸质量分数介于19.691～30.947 mg/g 之间,茶氨酸含量占游离氨基酸含量的55.1%～70.5%;随着3种茶园气温的降低,游离氨基酸含量和茶氨酸含量呈升高趋势,3种鲜味氨基酸（天冬氨酸、谷氨酸、茶氨酸）、5种甜味氨基酸（脯氨酸、丙氨酸、甘氨酸、苏氨酸、丝氨酸）和5种苦味氨基酸（异亮氨酸、亮氨酸、苯丙氨酸、酪氨酸、缬氨酸）共占游离氨基酸含量在防霜扇茶园和林间茶园中分别出现升高、降低和降低趋势,而在普通茶园中变化与之不同。分析认为：轻度“倒春寒”可导致无防护措施的普通茶园茶叶滋味劣变,林间茶园能够减轻冻害,防霜扇作为防护手段不仅可以减免冻害,还可避免茶叶滋味品质下降。该研究结果为后续深入研究不同防护措施茶园茶叶品质提供参考。     

Structure-activity relationships of tea compounds against human cancer cells

The content of the biologically active amino acid theanine in 15 commercial black, green, specialty, and herbal tea leaves was determined as the 2,4-dinitrophenyltheanine derivative (DNP-theanine) by a validated HPLC method. To define relative anticarcinogenic potencies of tea compounds and teas, nine green tea catechins, three black tea theaflavins, and theanine as well as aqueous and 80% ethanol/water extracts of the same tea leaves were evaluated for their ability to induce cell death in human cancer and normal cells using a tetrazolium microculture (MTT) assay. Compared to untreated controls, most catechins, theaflavins, theanine, and all tea extracts reduced the numbers of the following human cancer cell lines: breast (MCF-7), colon (HT-29), hepatoma (liver) (HepG2), and prostate (PC-3) as well as normal human liver cells (Chang). The growth of normal human lung (HEL299) cells was not inhibited. The destruction of cancer cells was also observed visually by reverse phase microscopy. Statistical analysis of the data showed that (a) the anticarcinogenic effects of tea compounds and of tea leaf extracts varied widely and were concentration dependent over the ranges from 50 to 400 microg/mL of tea compound and from 50 to 400 microg/g of tea solids; (b) the different cancer cells varied in their susceptibilities to destruction; (c) 80% ethanol/water extracts with higher levels of flavonoids determined by HPLC were in most cases more active than the corresponding water extracts; and (d) flavonoid levels of the teas did not directly correlate with anticarcinogenic activities. The findings extend related observations on the anticarcinogenic potential of tea ingredients and suggest that consumers may benefit more by drinking both green and black teas.     

Sensory activity,chemical structure,and synthesis of Maillard generated bitter-tasting 1-oxo-2,3-dihydro-1H-indolizinium-6-olates

Thermal treatment of aqueous solutions of xylose, rhamnose, and l-alanine led to a rapid development of a bitter taste of the reaction mixture. To characterize the key compounds causing this bitter taste, the recently developed taste dilution analysis (TDA), which is based on the determination of the taste threshold of reaction products in serial dilutions of HPLC fractions, was performed to locate the most intense taste compounds in the complex mixture of Maillard reaction products. By application of this TDA, 26 fractions were obtained, among which seven fractions were evaluated with a high taste impact. LC/MS and NMR spectroscopy as well as synthetic experiments revealed the 1-oxo-2,3-dihydro-1H-indolizinium-6-olates 1-5 as the key compounds contributing the most to the intense bitter taste of the Maillard mixture. Calculation of the taste impact of these compounds based on a dose/activity relationship indicated that these five compounds already accounted for 56.8% of the overall bitterness of the Maillard mixture, thus demonstrating this class of 1-oxo-2,3-dihydro-1H-indolizinium-6-olates as the key bitter compounds. First synthetic studies on the relationship between the chemical structure and the human psychobiological activity of 1-oxo-2,3-dihydro-1H-indolizinium-6-olates revealed that substitution of the furan rings of 1 by 5-methylfuryl moieties (compounds 3-5) or by 5-(hydroxymethyl)furyl groups (compound 6) led to a significant increase of the bitter threshold. In contrast, the substitution of the oxygen atoms in the furan rings of 1 by sulfur atoms induced a significant decrease of the detection threshold of the 1-oxo-2,3-dihydro-1H-indolizinium-6-olate; for example, the thiophene derivative 7 showed the extraordinarily low bitter detection threshold of 6.3 x 10(-5) mmol/kg (water).