Characterisation of volatile and non-volatile metabolites in etiolated leaves of tea (Camellia sinensis) plants in the dark

Aroma is an essential factor affecting the quality of tea (Camellia sinensis) products. While changes of volatile compounds during tea manufacturing have been intensively studied, the effect of environmental factors on volatile contents of fresh tea leaves has received less attention. We found that C. sinensis var. Yabukita kept in darkness by shading treatment for 3 weeks developed etiolated leaves with significantly increased levels of volatiles, especially volatile phenylpropanoids/benzenoids (VPBs). Upstream metabolites of VPBs, in particular shikimic acid, prephenic acid, and phenylpyruvic acid, showed lower levels in dark treated than in control leaves, whereas the contents of most amino acids including l-phenylalanine, a key precursor of VPBs, were significantly enhanced. In addition, analysis by ultra performance liquid chromatography-time of flight mass spectrometry, capillary electrophoresis–time of flight mass spectrometry, high performance liquid chromatography, and gas chromatography–mass spectrometry indicated that volatile and non-volatile metabolite profiles differed significantly between dark treated and untreated leaves.     

Effect of Steaming of Beans Pudding on the Phytochemical Composition of Thaumatococcus Daniellii Wrapper

The present study investigated the phytochemical analysis of Thaumatococcus daniellii leaves used as bean pudding wrapper in most part of Nigeria. The leaves were analysed before and after usage. The phytochemical components in the leaves were analysed qualitatively and quantitatively using standard methods. The toxicity of the leave extracts was determined by brine shrimp lethality assay and this recorded zero mortality. The TLC of the extracts was carried out and the chromatogram revealed differences in the number of components present in the fresh and used leaves. The amount of phytochemicals detected in the fresh leaves was higher than that of the used leaves e.g. 22.50% saponin was obtained in the methanol extract of the fresh leaf and 12.75% for used leaf. Tanin 8% in fresh leaf and 1.30% in used leaf of the methanol extracts. Some metabolites must have been absorbed by the bean pudding and some might have gone into the steam water.     

葡萄种质果实有机酸组分及其含量特性

为筛选优异葡萄种质,利用高效液相色谱对国家果树种质郑州葡萄圃保存的302 份葡萄种质果实开展葡萄有机酸组分及含量特性分析,并用聚类分析和主成分分析对葡萄果实中有机酸组分特征及含量规律进行总结及分析。结果表明：在302 份葡萄样品中,93.8%的样品酒石酸含量高于苹果酸,5.12%的样品苹果酸含量与柠檬酸相当（两者之差不大于2%）,另有7.37%的样品苹果酸含量高于酒石酸;栽培品种葡萄的酒石酸质量浓度大于苹果酸;野生种葡萄（除刺葡萄株系外）中,桑叶葡萄、腺枝葡萄、华东葡萄、河岸葡萄等果实中苹果酸质量浓度高于酒石酸,其质量浓度是酒石酸的1.2～1.7 倍。对比不同用途葡萄中有机酸质量浓度发现,酿酒葡萄的有机酸质量浓度明显高于其他用途葡萄,鲜食葡萄的pH值（3.776）显著高于其他用途葡萄。此外,对不同种群葡萄果实中有机酸质量浓度比较表明,东亚种群葡萄果实的有机酸质量浓度均高于其他种群,尤其是酒石酸的质量浓度（8.325 mg/mL）显著高于其他种群。样品间酒石酸、总酸和酸度值变异系数较小;样品间草酸、苹果酸和柠檬酸差异大;除pH值与草酸外,酒石酸与其他有机酸间均呈显著正相关;总酸质量浓度与酒石酸、草酸、苹果酸、柠檬酸质量浓度、可滴定酸含量、酸度值均呈显著正相关。野生种葡萄有机酸质量浓度远高于栽培种葡萄,尤其是中国特有的东亚种群山葡萄N43-3的有机酸质量浓度最高,因此可以当作有机酸质量浓度较高的优特异种质进行选育及利用。本研究结果可为葡萄种质资源的保护与利用提供一定理论依据。     

Stress metabolites from Corchorus olitorius L. leaves in response to certain stress agents

Five coumarins were found to be produced as phytoalexins (stress metabolites) from the fresh young leaves of Corchorus olitorius L. plant, in response to inoculation with biotic stress agent, such as the spore suspension of the fungus Helminthosporium turcicum and with chemical stress agents, such as aqueous solutions of mercuric chloride and cupric chloride. The five compounds were isolated, purified and subjected to melting point and spectroscopic determinations. They were identified as scopoletin, fraxinol, isopimpinellin, xanthotoxol and peucedanol. They were tested for their antimicrobial activities. The volatile components of the natural fresh leaves and the cupric chloride-treated leaves were prepared and subjected to GC/MS analysis. Fifty five and 49 components were identified in the control and treated leaves, respectively. cis-3-Hexen-1-ol, cis-4-hexen-1-ol, terpinolene, sabinene and phytol were the major compounds in the control leaves, while those of the treated leaves were cis-4-hexen-1-ol, cis-3-hexen-1-ol, tetradecanal and phytol. The percentage of the total oxygenated compounds were found to be increased in the cupric chloride-treated leaves much more than the control leaves (77.3 and 47.4%, respectively).     

Phenolic Compounds from the Leaves of Vitis labrusca and Vitis vinifera L. as a Source of Waste Byproducts: Development and Validation of LC Method and Antichemotactic Activity

The cultivation of Vitis (Vitaceae) grape varieties is one of the most important economic activities in agribusiness in southern Brazil. Vitis varieties are rich in polyphenolic compounds with several pharmacological and biological activities, such as antioxidant action. In this context, this study analyzed qualitatively and quantitatively the anthocyans and flavonoids found in the leaves of grape varieties Vitis vinifera and Vitis labrusca. For this purpose, vine leaf extracts were prepared and the chemical profile of each was characterized by LC/MS-MS. Two high performance liquid chromatography-validated methods were performed using UV/VIS-LC-DAD detector to quantify phenolic compounds. The main anthocyanins isolated from vine leaves were cyanidin-3-O-glucoside and peonidin-3-O-glucoside. The flavonoids identified were rutin, quercetin-3-O-galactoside, quercetin-3-O-glucoside, and quercetin-3-O-glucuronide, which was the predominant compound. The Waters X-Terra® RP18 column allowed the effective separation of quercetin-3-O-glucuronide from the other flavonoids for the first time, besides the partial separation of quercetin-3-O-galactoside from quercetin-3-O-glucoside. Furthermore, another phenolic compound was confirmed by MS spectrometry, using direct infusion, as being trans-caftaric acid. The present study also investigates the antichemotactic activity in vitro of grape crude extracts, fractions, and isolated compounds. It was demonstrated that almost all fractions and isolated compounds showed increased antichemotactic effect in response to LPS with a more pronounced values of IC₅₀ for anthocyanins fraction, rutin, quercetin-3-O-galactoside, and trans-caftaric acid (0.9, 1.6, 3.7, and 5.1 ng/mL, respectively).     

Gut microbial catabolism of grape seed flavan-3-ols by human faecal microbiota. Targetted analysis of precursor compounds, intermediate metabolites and end-products

In vitro batch culture fermentations were conducted with grape seed polyphenols and human faecal microbiota, in order to monitor both changes in precursor flavan-3-ols and the formation of microbial-derived metabolites. By the application of UPLC-DAD-ESI-TQ MS, monomers, and dimeric and trimeric procyanidins were shown to be degraded during the first 10 h of fermentation, with notable inter-individual differences being observed between fermentations. This period (10 h) also coincided with the maximum formation of intermediate metabolites, such as 5-(3′,4′-dihydroxyphenyl)-γ-valerolactone and 4-hydroxy-5-(3′,4′-dihydroxyphenyl)-valeric acid, and of several phenolic acids, including 3-(3,4-dihydroxyphenyl)-propionic acid, 3,4-dihydroxyphenylacetic acid, 4-hydroxymandelic acid, and gallic acid (5-10 h maximum formation). Later phases of the incubations (10-48 h) were characterised by the appearance of mono- and non-hydroxylated forms of previous metabolites by dehydroxylation reactions. Of particular interest was the detection of γ-valerolactone, which was seen for the first time as a metabolite from the microbial catabolism of flavan-3-ols. Changes registered during fermentation were finally summarised by a principal component analysis (PCA). Results revealed that 5-(3′,4′-dihydroxyphenyl)-γ-valerolactone was a key metabolite in explaining inter-individual differences and delineating the rate and extent of the microbial catabolism of flavan-3-ols, which could finally affect absorption and bioactivity of these compounds.     

八大香型风格新鲜烟叶代谢特征的生态成因分析

为考察生态因素对鲜烟叶代谢特征的影响,根据全国烤烟烟叶香型风格区划研究结果,按照八大香型风格,结合气象数据,分析了鲜烟叶特征代谢物(脂类化合物、含氮化合物、糖类化合物、有机酸类化合物、核酸类化合物、甾醇类化合物)与不同气象因子的相关性,并分析了各类代谢物的生态成因。结果表明,每个风格区的特征代谢物均与生态条件一定程度相关,温度、日照时数、降雨量等生态特征数据的变化可解释部分特征代谢产物的积累,表明鲜烟叶的代谢特征与环境生态条件紧密相关,其中与代谢物含量有较高相关性的气候因子为大田中后期温度和大田中后期日照时数。      

Effects of noble rot on must composition and aroma profile of Amarone wine produced by the traditional grape withering protocol

The effects of noble rot on the overall quality of Amarone wine, a “Passito” dry red wine, produced by a traditional withering grape process are poorly understood. Wines obtained from healthy and selected grapes infected by noble rot were compared. Validation of the experimental plan was carried out by determining glycerol and gluconic acid content, laccase activity and microbial analysis of the musts. Botrytis cinerea infection produced significant variation in esterase and β-glucosidase activity of grape must. Aroma analysis in healthy and botrytised wines was carried out by SPE extraction followed by GC–MS quantification. A decrease of several fermentative esters was observed in botrytised wine, while wine produced from healthy grapes showed a higher fatty acid content. Molecules such as 1-octen-3-ol, phenylacetaldehyde and furaneol were positively linked to the noble-rotten grapes. Interestingly, γ-nonalactone and 4-carbethoxy-γ-butyrolactone increased, while sherry lactones decreased in botrytised wine. The strong increase of N-(3-methylbutyl)acetamide was related to B. cinerea grape infection.     

Effect of environmental factors [air and UV-C irradiation] on some fresh fruit juices

The present study was carried out to evaluate the effect of environmental factors (air and UV-C irradiation) on various common fruits juices (lemon, orange, papaya and grape). Total phenolics, total ascorbic acid (AA) content and antioxidant capability were determined in atmospheric air oxidation on fresh fruit juices, and the effect of UV-C irradiation on AA was investigated in both fresh fruit juices and commercial AA using RP-HPLC. The total phenolics content was found to be 92.46?±?1.5, 41.53?±?2.1, 138.56?±?2.3 and 110.54?±?3.9?mg/100?ml of lemon, orange, papaya and grape juices, respectively. DPPH radical–scavenging activity of the fresh juices was 94.53?±?3.1, 82.36?±?2.2, 94.65?±?3.5 and 90.74?±?2.6 percentage, whereas the 1-day-old fruit juices showed reduced scavenging activity when compared to fresh juices. Total phenolic (TP) and DPPH-scavenging activity were significantly decreased in the mean at P?<?0.05 level. Ascorbic acid (vitamin C) content of fresh fruit juices was determined through RP-HPLC; fruit juices that were kept in open container for 24 hrs and that were treated with UV irradiation have shown a decrease in AA content, when compared with control fresh juice samples. AA degradation increased with high significant differences at P?<?0.001 level, with the increase in oxidation time in atmospheric air and UV-C radiation.     

植物中原花青素含量的分析测定

通过对多种植物的花、叶、皮、壳、籽、核等部位的原花青素含量进行分析测定。结果表明:原花青素广泛分布于植物体的不同部位中,原花青素除葡萄籽含量较高以外,在某些植物的叶和花中含量也较丰富,葡萄籽中原花青素以新鲜葡萄籽含量最高,经过酿酒和放置时间的延长原花青素含量降低。     

First steps towards the development of a non-destructive technique for the quality control of wine grapes during on-vine ripening and on arrival at the winery

NIR spectroscopy was used as a non-destructive technique for the assessment of changes in certain internal quality properties of wine grapes (Vitis vinifera L.) during on-vine ripening and at harvest. A total of 108 different wine grape samples were used to construct calibration models based on reference data and NIR spectral data, obtained using a commercially-available diode-array spectrophotometer (380-1700 nm). The feasibility of testing bunches of intact grapes was investigated and compared with more traditional methods of presentation, such as berries or must. Predictive models were constructed to quantify changes in soluble solid content (SSC, °Brix), reducing-sugar content (g/l), pH-value, titrable acidity (g/l tartaric acid), tartaric acid (g/l) and malic acid (g/l), these being the major parameters used to chart ripening. NIRS technology provided good precision for the bunch analysis mode assayed for SSC (r² = 0.89; SECV = 1.41 °Brix), for reducing-sugar content (r² = 0.87; SECV = 17.13 g/l) and for pH-value (r² = 0.69; SECV = 0.19). Models developed for testing other fruit acidity parameters yielded results sufficient to provide a screening tool to distinguish between low and high acidity values in intact grapes. Significantly, the results obtained with bunch presentation were similar to those obtained with berries and must, thus justifying further implementation of NIRS technology for the non-destructive analysis of quality properties both during on-vine ripening and on arrival at the winery. This method allows musts to be processed separately depending on initial grape quality, assessed with a single spectrum measurement and in a matter of seconds.     

江西茶鲜叶中重金属状况多变量分析

对江西省5个茶区的88个茶鲜叶(新叶、老叶)样,以国标方法测定重金属Cu、Pb、Cd、As、Hg、Cr含量。结果显示,老叶、新叶和鲜叶聚类分析结果不完全相同,但均分为5组,无明显自然地域界限,6种重金属聚类组间差异显著(P<0.05);经交叉验证的判别分析,区分老叶与新叶的正确率为87.5%,说明老叶与新叶差异明显,判别新叶是否高山组的正确率为77.3%。以改进的Nemerow综合污染指数(NIPI)评价重金属状况并进行简单对应分析显示,老叶主要为轻度污染,新叶总体未污染,处于警戒水平:赣中赣南茶区新叶安全,其余新叶为警戒水平。结论表明:江西茶鲜叶重金属茶区间有一定差异但自然地域界限不明显;种植环境平均良好,路边组与高山组无明显差异。     

Physico-chemical properties of cell wall materials obtained from ten grape varieties and their byproducts: grape pomaces and stems

The availability and the potential of wine by-products, grape pomaces and stems, obtained from ten different grape (Vitis vinifera L.) varieties (six red and four white) as raw materials for the production of dietary fibre (DF) concentrates were evaluated. The overall chemical composition, carbohydrate analysis and functional properties of DF from fresh grapes, grape pomaces and stems from those grape varieties were analysed. Both winemaking by-products presented considerable quantities of DF, ranging from 60% to 90% of total dry matter. The cell wall polysaccharides (CWP) composition of grape pomaces and fresh grapes was rather similar, pectic substances being the main component of the cell walls (40–54% total CWP). In contrast, cellulose was the predominant cell wall polymer for the stems (40–49% total CWP). In addition, the pectin content of grape pomaces exhibited significant differences between red and white grape varieties. The degree of methyl-esterification of uronic acids from wine by-products ranged from 21 to 39%; thus, pectic polysaccharides from the different samples could be classified as low methyl-esterified pectins. Klason lignin accounted for around 20–25% of DF in both grape pomaces and stems. With regard to the potential incorporation of DF concentrates into the food chain, functional properties were also determined. Swelling, water and fat retention capacity results showed great variability depending on the grape variety analysed.     

不同培养基的平菇多酚及氨基酸含量分析

该研究将不同粉碎处理的葡萄籽和朝鲜蓟叶添加在培养基中,主要研究了工业副产物对平菇营养物质及活性成分的影响。以总酚含量、总黄酮含量、DPPH、ABTS自由基清除能力及Fe3+还原能力为检测指标对其抗氧化活性进行评估;此外,利用相关性分析、主成分分析和偏最小二乘判别等分析手段对平菇氨基酸组成进行探究。结果表明,培养基中添加葡萄籽粉和朝鲜蓟叶粉,其总酚、总黄酮含量及抗氧化活性均高于CK组,尤其以氨基酸组成及含量显著增加,且氨基酸高贡献组分相对较多,前两个主成分可解释的累计方差达94.4%,其中对A1样品高贡献率的氨基酸有Ala、Tyr、Lys、Ser、Thr、Asp、Ary、Glu、Phe和His;对A2高贡献率氨基酸包括Ile、Val、Gly、Pro和Leu,而CK组不存在高贡献率的氨基酸组分。因此,葡萄籽和朝鲜蓟叶可代替传统棉籽壳培养基培育平菇,有助于平菇营养物质的富集及活性成分的提高。这些结果为工业副产物的综合利用提供理论依据,具有一定的实际应用价值和意义。     

Antioxidant properties and bioactive components of Norton (Vitis aestivalis) and Cabernet Franc (Vitis vinifera) wine grapes

Three wine grapes, Norton (Vitis aestivalis), Cabernet Franc clone1, and Cabernet Franc clone313 (Vitis vinifera), collected from a Virginia vineyard were evaluated and compared for their antioxidant properties and phenolic profile. All grape extracts exerted remarkable antioxidant activities. Their oxygen radical absorbance capacity (ORAC) values were not significantly different from one another, ranging from 22.9 to 26.7 μmol TE/g of fresh weight. The Cabernet Franc clone1 had the strongest 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals scavenging activity (8.8 μmol TE/g) compared to the Norton or Cabernet Franc clone313 grape extracts (7.9 μmol TE/g and 5.4 μmol TE/g, respectively). The Norton grape contained significantly higher total phenolic, anthocyanin, and flavonoid content than the Cabernet Franc grapes (p < 0.05). The hydroxybenzoic acids, in particular gallic acid, were the major phenolic acids in all the grape extracts. The Norton grape variety was found to be rich in malvidin-diglucoside and malvidin-glucoside, while the malvidin-diglucoside was negligible in the Cabernet Franc grapes. The results suggest a remarkable impact of grape genotype on its antioxidant properties and phenolic composition in Virginia-grown wine grapes.     

Effects of phenanthrene on chemical composition and enzyme activity in fresh tea leaves

Phenanthrene pollution around roots of tea (Camellia Sinensis) seedlings and its effects on antioxidative enzymes and chemical composition in fresh tea leaves during 7 days of hydroponic culture were investigated. Phenanthrene concentration in the fresh tea leaves gradually increased over the 7 days. The activities of polyphenol oxidase (PPO) and superoxide dismutase (SOD) were promoted at the beginning of phenanthrene contamination and were suppressed from the fifth day, while peroxidase (POD) and catalase (CAT) followed an opposite trend. On the third day, the contents of water extract, amino acid and caffeine had peak values, and the protein content was the lowest. The contents of polyphenols and total sugar were increased on the first day and then began to decline. On the seventh day, all chemical components decreased, indicating that phenanthrene may reduce tea quality. Linear correlation analysis showed that the activities of PPO, POD and CAT and the content of polyphenols in the tea leaves correlated well with the concentration of phenanthrene in the tea leaves.     

基于非靶向代谢组学的红花大金不同生育期烟叶代谢图谱差异分析

  目的  为分析不同生育期烟草烟叶代谢图谱差异。  方法  以烟草品种红花大金元为研究对象,采集团棵期、现蕾期和成熟期烟叶样品,利用LC-QTOF/MS进行代谢指纹图谱检测,利用统计分析方法对差异代谢物进行筛选,基于Spearman相关分析计算代谢物之间的相关性系数,利用MetaboAnalyst软件进行代谢通路富集分析。  结果  （1）不同生育期烟草样品代谢图谱存在明显差异,共鉴定出245个差异代谢物,其中类黄酮、有机酸、氨基酸、核苷、生物碱、脂类等物质种类最多。（2）烟碱及其衍生物,如烟碱-1-氧代、麦斯明、假木贼碱、马钱子碱、新烟碱、2, 3'-二联吡啶,均随烟草生育期而逐渐累积,而与烟碱合成相关的前体物质,如精氨酸、亚精胺、腐胺、羟基丁酸等,均随着生育进程逐渐减少,且与烟碱含量显著负相关。（3）与次级代谢物合成相关氨基酸和有机酸等物质在团棵期或现蕾期含量较多,而在成熟期含量较低,与成熟期次级代谢物合成有关。（4）大部分类黄酮和酚酸组分在成熟期时含量较高,可以提高烟草抗病和抗逆能力。（5）相关性分析发现,烟碱含量与谷氨酸、泛酸、吡哆醛、蜜二糖、3-羟基邻氨基苯甲酸和顺式玉米素-o-葡萄糖苷等物质显著正相关,可能是调控烟叶中烟碱含量的重要标志物。  结论  基于代谢组学揭示了不同生育期烟草代谢物变化规律,为优质烟草育种及品质分析提供技术支撑。      

Chicoric acid levels in commercial basil (Ocimum basilicum) and Echinacea purpurea products

Fresh basil (Ocimum basilicum) leaves contain chicoric acid, which is the principal phenolic compound in Echinacea purpurea and purportedly an active ingredient in dietary supplements derived from E. purpurea. Here the concentrations of chicoric acid in dried and fresh basil products available to consumers, and how these concentrations compare to those from E. purpurea are reported. A wide range of chicoric acid concentrations (6.48–242.50 mg/100 g or 100 mL) were found in the dried basil flakes, fresh basil leaves, E. purpurea extracts, and E. purpurea capsules. Fresh basil leaves had higher concentrations of chicoric acid than dried basil flakes. Although E. purpurea extracts and capsules contained higher concentrations of chicoric acid than fresh basil leaves, basil could be an economical and more readily available source for chicoric acid for consumers. Additionally, cultivar selection, dehydration processing improvements, and proper storage methods may improve the final chicoric acid levels of future basil crops and products.     

Lactic acid bacteria community dynamics and metabolite production of rye sourdough fermentations share characteristics of wheat and spelt sourdough fermentations

Four spontaneous rye sourdough fermentations were performed over a period of ten days with daily back-slopping. Samples taken at all refreshment steps were used for culture-dependent and culture-independent characterization of the microbiota present. Furthermore, an extensive metabolite target analysis was performed through a combination of various chromatographic methods, including liquid chromatography coupled to mass spectrometry (LC/MS) and gas chromatography coupled to mass spectrometry (GC/MS). Spearman’s rank correlation coefficients were calculated and a principal component analysis (PCA) was performed on the data obtained in this study combined with data obtained previously for wheat and spelt sourdoughs. In general, the establishment of a stable microbial ecosystem occurred through a three-phase evolution, with mainly Lactobacillus plantarum and Lactobacillus fermentum dominating the rye sourdough ecosystems. PCA revealed that ornithine and mannitol were positively correlated with rye sourdoughs, contributing to bacterial competitiveness at the onset of sourdough production. Wheat and spelt sourdoughs showed a high degree of similarity, although certain compounds (e.g. indolelactic acid) appeared to be specific for spelt sourdoughs. The production of amino acid metabolites, mainly hydroxy acids (e.g. phenyllactic acid) and alcohols (e.g. 3-methyl-1-butanol), contributed to the equilibration of the redox balance and further enhanced the competitiveness of dominant species in stable sourdoughs.     

Discrimination of young and mature leaves of Melicope ptelefolia using H NMR and multivariate data analysis

The ‘Ulam’, a traditional Malay dish, are plants that can be eaten raw, as a form of local salad. The shoots and young leaves of Melicope ptelefolia are among the popular species, believed to be high in nutritional and medicinal values. The metabolomic fingerprinting analysis of the ethanolic extracts of leaves of M. ptelefolia was carried out using ¹H Nuclear Magnetic Resonance (NMR) spectroscopy and multivariate data analysis in order to differentiate young and mature leaves and to evaluate the variation of their chemical composition. Principle component analysis (PCA) of the ¹H NMR spectra showed a clear discrimination between the young and mature leaves extracts by PC3 and PC4. The compounds responsible for the differentiation were identified by comparison of ¹H NMR chemical shifts and qualitative HPLC. The young leaves were found to be richer in fatty acids and the levels of the three marker compounds, p-O-geranylcoumaric acid, 2,4,6-trihydroxy-3-geranylacetophenone and 2,4,6-trihydroxy-3-prenylacetophenone, were clearly higher. The mature leaves contain higher levels of sugars and glycosidic components.