基于化学成分的古树普洱茶原料产地鉴别研究

目的基于化学成分鉴别古树普洱茶原料的产地。方法测定不同产地不同季节古树普洱茶原料中主要化学成分,并对数据进行差异性分析、主成分分析、Fisher判别分析。结果春季古树普洱茶原料的水浸出物、茶多酚含量较高,秋季古树普洱茶原料的粗纤维、游离氨基酸总量、可溶性糖、儿茶素总量较高;不同产地古树普洱茶原料在茶多酚、儿茶素(catechin,C)、表儿茶素(epicatechin,EC)、表没食子儿茶素(epigalloeateehin, EGC)、表儿茶素没食子酸酷(epieateehingallate, ECG)存在显著差异;运用主成分分析可以很好地区分不同季节的古树普洱茶原料,但是不能很好区分不同产地的古树普洱茶原料;采用Fisher判别分析方法可以较好地区分不同产地的普洱茶原料,总判别正确率为90%。结论茶叶中的化学成分可以用于区分不同季节不同产地的古树普洱茶原料。     

基于矿质元素的古树普洱茶原料特征研究

目的基于矿质元素鉴别古树普洱茶原料的产地。方法测定不同产地不同季节古树普洱茶原料中矿质元素,并对数据进行了单因素分析、主成分分析、系统聚类分析、Fisher判别分析。结果不同产区的古树普洱茶原料矿质元素:普洱茶区锌、钠、钙、铬含量较高,版纳茶区的锰、铜、钾含量较高,临沧茶区的铁、氟、稀土总量较高;不同产地古树普洱茶原料铁、钙存在极显著差异。春季古树普洱茶原料的铁、锰、锌、钾、钠、钙、镁、稀土总量较高,秋季古树普洱茶原料的铜、铬、氟含量较高,不同季节古树普洱茶原料铁、铜、钾、钠、钙、铬含量存在极显著差异。运用主成分分析可以很好地区分不同季节的古树普洱茶原料,但是不能很好区分不同产地的古树普洱茶原料;采用系统聚类分析方法不能区分不同产地古树普洱茶原料;采用Fisher判别分析方法可以较好地区分不同产地的普洱茶原料,总判别正确率为86.7%。结论茶叶中的矿质元素可以用于区分不同季节不同产地的古树普洱茶原料。     

云南固态速溶普洱茶生产企业标准现状分析

目的了解速溶普洱茶行业现状,为规范固态速溶普洱茶产业,制定固态速溶普洱茶行业标准提供市场依据。方法通过对固态速溶茶国家标准与55个速溶普洱茶生产企业60个企业标准理化指标和安全性指标的综合分析,确定各理化指标的基本数值范围和企业标准涉及的安全性指标种类,并对国家标准与相关企业标准进行对比。结果水分含量在8%~15%的企业标准数量占调查企业标准总数的61.36%,总灰分含量在15%~25%的企业标准数量占调查企业标准总数的78.82%,茶多酚含量在10%~20%的企业标准数量占调查企业标准总数的69.35%;安全性指标可参考国家相关标准。结论固态速溶普洱茶行业标准可结合现有固态速溶茶国家标准和产业实际需求进行制定。     

The analysis by HPLC of green,black and Pu'er teas produced in Yunnan

Samples of tea (Quingmao, steamed, black and Pu'er) produced in Yunnan Province, People's Republic of China, from the locally characteristic large-leaved Camellia sinensis, have been brewed and the brews analysed for their contents of flavonol glycosides, flavan-3-ols and their oxidation products. Several samples of Pu'er tea manufactured in the 1920s, 1930s and 1940s were also examined. The brews prepared from Quingmao tea and steamed green tea leaf were very similar containing primarily flavan-3-ols accompanied by flavonol glycosides, theogallin and gallic acid. The profile of flavanols was unusual, being dominated by (?)-epicatechingallate (rather than (?)-epigallocatechingallate) and by an unusually large (+)-catechin content. The brew of the equivalent black tea retained significant amounts of residual flavanols (particularly (?)-epicatechingallate and (+)-catechin), and contained significant amounts of thearubigins, theaflavic acids and theaflavins (including the less common analogues probably derived from the oxidation of (+)-catechin). The brews prepared from the Puer teas were distinguishable from the black tea brew by the absence of theaflavins and theaflavic acids, and by much smaller residues of flavanois and flavonol glycosides. The contents of thearubigins varied considerably whereas those of gallic acid were similar.     

茶叶提取物体外抗氧化活性与其功能性成分含量的相关性研究

分别选取信阳毛尖、红碎茶、普洱熟茶为典型不发酵、全发酵及后发酵茶种,测定其醇提物及水提物中的主要成分,并以氧自由基的吸收能力(ORAC)和DPPH自由基的清除能力为指标,比较各样品体外抗氧化活性,采用功能化合物重组模型,探究茶叶提取物抗氧化活性的主要贡献物质。结果表明:茶叶醇提物中茶多酚、咖啡因及醇溶蛋白的含量较高,其ORAC值及DPPH自由基清除能力均较对应水提物强;茶叶水提物功能性组分以茶多酚、茶多糖及氨基酸为主,都具有一定的抗氧化活性。其中茶多酚含量,尤其是酯型儿茶素(ECG和EGCG)含量是影响茶叶提取物抗氧化活性的主要因素。重组试验结果表明,儿茶素和没食子酸是不发酵茶(信阳毛尖)提取物的主要抗氧化组分,但并不是红碎茶、普洱熟茶等发酵茶中的主要抗氧化成分。     

Isolation and identification of spermidine derivatives in tea (Camellia sinensis) flowers and their distribution in floral organs

BACKGROUND: Recently, tea (Camellia sinensis) flowers have attracted increasing interest because of their content of bioactive compounds such as catechins. The aim of this study was to investigate the occurrence of some characteristic compounds in tea flowers. RESULTS: A principal component analysis of metabolites using ultra‐performance liquid chromatography/time‐of‐flight mass spectrometry showed differences in metabolite profile between flowers and leaves of C. sinensis var. Yabukita. Four spermidine derivatives were isolated from tea flowers. One of them was determined as N¹,N⁵,N¹⁰‐tricoumaroyl spermidine based on NMR, MS and UV data. The other three were identified as feruoyl dicoumaroyl spermidine, coumaroyl diferuoyl spermidine and triferuoyl spermidine based on MSⁿ data. Tricoumaroyl spermidine as the major spermidine conjugate was not detected in tea leaves. Furthermore, it decreased during floral development and mainly occurred in anthers. CONCLUSION: This study has provided the first evidence that spermidine‐phenolic acid conjugates occur in tea flowers in considerable amounts. Their presence should prompt a reconsideration of the ecological role of tea flowers. From an economic point of view, tea flowers might be suitable as a raw material in the healthcare food and pharmaceutical industries. Copyright © 2012 Society of Chemical Industry     

Characterizing Soy Sauce Moromi Manufactured by High‐Salt Dilute‐State and Low‐Salt Solid‐State Fermentation Using Multiphase Analyzing Methods

Present study was to characterize the physiochemical properties, free amino acids (FAAs), volatiles and microbial communities of various moromi, respectively sampled from different stages of high‐salt dilute‐state (HSDS) and low‐salt solid‐state (LSSS) fermentation, using multiphase analyzing methods. Phospholipid fatty acids (PLFA) analysis indicated that Gram‐positive bacteria were dominant bacteria and fungi were principal microbes. For DGGE analysis, dominant microbes in moromi were mainly fell into Weissella, Tetragenococcus, Candida, Pichia, and Zygosaccharomyces. During fermentation, the dominant microbes shifted from nonhalophilic and less acid‐tolerant species to halophilic and acid‐tolerant species. Total of 15 FAAs and 44 volatiles were identified in moromi, mainly Glu, Asp, Tyr, and acids, alcohols, esters, aldehydes, respectively. Odor activity values analysis suggested that the final moromi of LSSS fermentation had more complicated odors than that of HSDS fermentation. Conclusively, technological parameters, microbial communities, raw materials and fermentation process may result in the discrepancy of HSDS and LSSS moromi.     

Bacterial and fungal diversity in the traditional Chinese strong flavour liquor Daqu

In this study, the diversity of bacterial and fungal communities in the strong flavour Daqu production process was investigated using polymerase chain reaction denatured gradient gel electrophoresis (PCR‐DGGE). The band number, dominance, diversity and similarity of the 16S rDNA differed in the DGGE patterns, reflecting the wide diversity of the microbial communities present in the different phases. The band abundance and dominance of the eukaryotic microbial community varied in the different phases. To provide insight into factors contributing to this diversity, the physicochemical properties of Daqu were explored. The core temperature of Daqu gradually increased to a maximum during the Dinghuo phase followed by a decline. The moisture and starch content decreased throughout fermentation. The total acid, reducing sugar, amino nitrogen, α‐amylase activity, glucoamylase activity, protease activity, bacterial and mould number increased gradually to a maximum value at the end of the Peijun phase before decreasing in the Dinghuo phase increasing slightly in the Huanluo phase and then remaining almost unchanged. The esterase activity exhibited a similar change to that of other enzymes and then slightly increased after the Peijun stage. For yeasts, the highest number was obtained at the Peijun stage and then decreased. Correlation analysis revealed that the diversity of both bacteria and fungi was negatively correlated with moisture and starch and positively correlated with other indices. The reducing sugar, amino nitrogen, α‐amylase and protease activities showed significant positive correlation with bacteria, and glucoamylase and esterase showed significant correlation with fungi. © 2019 The Institute of Brewing & Distilling     

Molecular Identification of Microbial Community in Surface and Undersurface Douchi During Postfermentation

To find the reason for fermentation failure of surface Douchi during postfermentation, the microbial communities in undersurface and surface samples were investigated using cell counting method and denaturing gradient gel electrophoresis (DGGE). The results showed that the microbial biomass in surface Douchi was obviously different from that in undersurface Douchi even sampled from the same fermentation tanks, and a 10‐ to 100‐fold reduction of microbial cell counts in undersurface had been observed. The bacterial DGGE profile and principal component analysis (PCA) results indicated that only Lactococcus lacts subsp. lactis and Bacillus thermoamylovorans were detected from surface Douchi, while Lactococcus lacts subsp. lactis, Staphylococcus lentus and 2 uncultured strains occupied the dominant positions in undersurface Douchi; when amplified using Bacillus‐specific primers, Bacillus thermoamylovorans, Bacillus subtilis, and Enterobacter sp. were found in undersurface Douchi, while only Bacillus thermoamylovorans were detected from surface Douchi; compared to the bacteria and Bacillus, the DGGE profiles and PCA plot of fungi indicated that the fungal community between surface and undersurface Douchi was similar and mainly composed by yeasts. In this study, we detected the microbial biomass and species in postfermentation stage of Douchi, and the various microbial diversity in undersurface and surface samples might be the cause of the fermentation failure in surface fermentation tanks.     

The Occurrence of Beer Spoilage Lactic Acid Bacteria in Craft Beer Production

Beer is one of the world's most ancient and widely consumed fermented alcoholic beverages produced with water, malted cereal grains (generally barley and wheat), hops, and yeast. Beer is considered an unfavorable substrate of growth for many microorganisms, however, there are a limited number of bacteria and yeasts, which are capable of growth and may spoil beer especially if it is not pasteurized or sterile‐filtered as craft beer. The aim of this research study was to track beer spoilage lactic acid bacteria (LAB) inside a brewery and during the craft beer production process. To that end, indoor air and work surface samples, collected in the brewery under study, together with commercial active dry yeasts, exhausted yeasts, yeast pellet (obtained after mature beer centrifugation), and spoiled beers were analyzed through culture‐dependent methods and PCR‐DGGE in order to identify the contaminant LAB species and the source of contamination. Lactobacillus brevis was detected in a spoiled beer and in a commercial active dry yeast. Other LAB species and bacteria ascribed to Staphylococcus sp., Enterobaceriaceae, and Acetobacter sp. were found in the brewery. In conclusion, the PCR‐DGGE technique coupled with the culture‐dependent method was found to be a useful tool for identifying the beer spoilage bacteria and the source of contamination. The analyses carried out on raw materials, by‐products, final products, and the brewery were useful for implementing a sanitization plan to be adopted in the production plant.     

Bacterial Community Dynamics of Salted and Fermented Shrimp based on Denaturing Gradient Gel Electrophoresis

The Korean traditional seafood jeotgal is consumed directly or as an additive in other foods to improve flavor or fermentation efficiency. Saeujot, made from salted and fermented tiny shrimp (SFS; Acetes japonicus), is the best‐selling jeotgal in Korea. In this study, we reveal the microbial diversity and dynamics in naturally fermented shrimp by denaturing gradient gel electrophoresis (DGGE). The population fingerprints of the predominant microbiota and its succession were generated by DGGE analysis of universal V3 16S rDNA polymerase chain reaction (PCR) amplicons. Overall, 17 strains were identified from sequencing of 30 DGGE bands. The DGGE profiles showed diverse bacterial populations in the sample, throughout the fermentation of SFS. Staphylococcus equorum, Halanaerobium saccharolyticum, Salimicrobium luteum, and Halomonas jeotgali were the dominant bacteria, and their levels steadily increased during the fermentation process. Certain other bacteria, such as Psychrobacter jeotgali and Halomonas alimentaria appeared during the early‐fermentation process, while Alkalibacterium putridalgicola, Tetragenococcus muriaticus, and Salinicoccus jeotgali appeared during the late‐fermentation process. The members of the order Bacillales were found to be predominant during the fermentation of SFS. Furthermore, S. equorum was identified as the dominant bacterial isolate by the traditional method of culturing under aerobic and facultative anaerobic conditions. We expect that this information will facilitate the design of autochthonous starter cultures for the production of SFS with desired characteristic sensory profiles and shorter ripening times.     

Layer‐by‐Layer Thin Film of Iron Phthalocyanine as a Simple and Fast Sensor for Polyphenol Determination in Tea Samples

Polyphenols have attracted attention due to their antioxidant capacity and beneficial effects to health. Therefore, fast, inexpensive, and efficient methods to discriminate and to quantify polyphenols are of interest for food industry. In this paper, Layer‐by‐Layer films of poly(allylamine hydrochloride) and iron tetrasulfonated phthalocyanine were employed as sensor for determination of polyphenols in green tea (camellia sinensis), and green and roasted mate teas (ilex paraguariensis). The polyphenol sensor was tested in catechol standard solution by differential pulse voltammetry (DPV), reaching a limit of detection of 1.76 × 10^?7 mol/L. The determination of polyphenols in the tea samples was obtained by analytical curve and catechol standard addition using electrochemical techniques. Projection techniques (information visualization) were applied to the DPV results of the tea samples and a pattern of separation following the phenolic content was obtained. The results support the application of the sensor in fast classification of beverages according to their polyphenol content.     

普洱茶贮藏过程中“金花”菌的鉴定及其对茶叶品质的影响

由于仓储环境的差异,普洱茶在贮藏过程中偶尔会产生"金花"。以金花普洱茶(熟茶)为实验材料,并以同批次普通普洱茶(熟茶)为参照,对水浸出物、茶多酚、游离氨基酸、茶色素(茶黄素、茶红素、茶褐素)、咖啡碱、水溶性糖、儿茶素类等化学成分含量进行测定;采用平板划线法对"金花"菌进行分离和菌落数量测定,并结合形态观察和分子生物学方法对"金花"菌进行鉴定。结果表明:金花普洱茶(熟茶)和普通普洱茶(熟茶)在水浸出物、茶多酚、儿茶素总量、EC、EGC、茶黄素、游离氨基酸、咖啡碱含量上无显著性差异(P>0.05)。金花普洱茶(熟茶)中茶红素、水溶性总糖、没食子酸含量显著(P<0.05)高于普通普洱茶(熟茶),EGCG、茶褐素含量略低于普通普洱茶(熟茶);在普洱茶贮藏过程中分离出3株"金花"菌,分别鉴定为Aspergillus pallidofulvus(相似度99.9%)、Aspergillus sesamicola(相似度99.8%)、Penicillium manginii(相似度99.6%)。其中Aspergillus sesamicola为优势"金花"菌种,占"金花"菌群数量总量的66%。首次对普洱茶贮藏中的"金花"菌进行了鉴定,初步分析了"金花"菌对普洱茶(熟茶)品质的影响,为"金花"菌在普洱茶中的应用提供了一定的理论基础与参考价值。     

Analysis of the Fungal Community in Zaopei During the Production of Chinese Luzhou‐flavour Liquor

Denaturing gradient gel electrophoresis (DGGE) and 18S rRNA gene analysis were carried out to analyze the fungal community in Zaopei during the production of Chinese Luzhou‐flavour liquor. Two pairs of primers (EF4/Fung5 and EF4/NS2‐GC) were used for the DGGE. Polymerase chain reaction (PCR) for clone analysis was preformed with the primers (EF4/EF3). The results of the DGGE analysis showed that the fungal 18S rRNA genes taken from the Zaopei sample of the pit had high diversity. Gene clone libraries containing 120 clonal sequences of 18S rRNA were constructed. The fungal diversity shift showed that the fungal genera changed with increasing fermentation time and four genera of fungi (Issatchenkia, Talaromyces, Aspergillus and Eurotium) were the main dominant communities during the fermentation of Chinese Luzhou‐flavour liquor. The clonal sequences of the genera Rhizopus and Saccharomyces were found with difficulty from the clone libraries of Zaopei, suggesting that the genera Rhizopus and Saccharomyces perhaps are not necessary for saccharifying and fermenting grains during the production of Chinese Luzhou‐flavour liquor.     

Structural characterization and antioxidant activities of 2 water-soluble polysaccharide fractions purified from tea (Camellia sinensis) flower

Abstract: The water‐soluble crude polysaccharide tea flower polysaccharide (TFP), obtained from tea (Camellia sinensis) flower by boiling‐water extraction and ethanol precipitation, was fractionated by Sephadex G‐100 column chromatography, giving 2 polysaccharide fractions termed TFP‐1 and TFP‐2. The structural features of TFP‐1 and TFP‐2 were investigated by high‐performance liquid chromatography (HPLC), gel‐permeation chromatography (GPC), rheometer, infrared (IR) spectra, nuclear magnetic resonance (NMR) spectroscopy, atomic force microscope (AFM), and scanning electron microscope (SEM). Results indicated that TFP‐1 was composed of glucose: xylose: rhamnose: galactose = 1.0:1.2:0.81:0.98 with a molecular weight of 167.5 KDa, while TFP‐2 comprised glucose: xylose: rhamnose: arabinose = 1.0:0.76:2.3:2.3 with a molecular weight of 10.1 KDa. The ¹H NMR revealed that TFP‐1 contained α‐L‐Rhap, α‐D‐Galp, α‐D‐GalpNAc, α‐D‐Xylp, α‐D‐Glcp, and β‐D‐Glcp residues, while TFP‐2 was illustrated to have α‐L‐Rhap, α‐L‐Arap, α‐D‐Xylp, α‐D‐Glcp, and α‐D‐GlcpNAc residues. Antioxidant activities of these fractions were investigated using various in vitro assay systems compared with ascorbic acid. In conclusion, TFP‐2 exhibited the higher antioxidant activities and could be explored as a novel potential antioxidant. Practical Application: At present, commonly low‐grade tea is preferred to extract the tea polysaccharide, to take full advantage of tea flower resource to extract polysaccharides can greatly reduce the cost of tea products. Thus, the search for plant‐derived biomaterials from this study could generate natural value‐added products from underutilized tea plant waste and used as a medicinal agent against chronic health problems, such as cancers, aging, and atherosclerosis caused by reactive free radicals that produced from oxidation.     

Effect of withering on the polyphenol oxidase level in the tea leaf

The level of polyphenol oxidase in tea leaf (Camellia sinensis) decreased on partial desiccation of the leaf during the withering process of black tea manufacture. However, sufficient activity remained, even after withering, to oxidise the available substrate in the leaf. The loss in activity due to desiccation was restored on rehydration.     

Microbiological characterisation of Robiola di Roccaverano cheese using PCR-DGGE

The aim of this study was the microbiological characterisation of a typical Italian cheese “Robiola di Roccaverano” with Protected Designation of Origin (PDO). Fresh and ripened robiola samples were collected from four artisanal and one industrial producer. Artisanal producers used raw goat's milk and natural fermentation, whilst the industrial producer used mixed cow–goat's milk and selected starters. The microbial communities were monitored during different seasons and ripening times with PCR–DGGE and culture-dependent methods.The cluster analysis showed that the DGGE bacterial patterns were related to the different manufacturing and climatic conditions, revealing the occurrence of species associated to Robiola di Roccaverano. The DGGE profiles of yeasts were affected by ripening in summer season.Moreover, the results obtained allowed the identification of microbial species such as Lactococcus lactis subsp. lactis, Geotricum spp. and Kluyveromyces lactis that are related to the production of this typical cheese.     

Determination of the survival kinetics of Salmonella spp. on the surface of ripened raw milk cheese during storage at different temperatures

The aim of this study was to determine the survival kinetics of Salmonella enterica subsp. enterica in ripened raw milk cheese. Cheese samples inoculated with S. enterica subsp. enterica were stored at 5, 15 and 25 °C and analysed in terms of physico-chemical and microbiological characteristics. Three primary models were used to estimate the kinetic parameters of S. enterica subsp. enterica. The secondary Arrhenius model was used to establish the relationship between temperature and parameter α of the Weibull model. Additionally, prediction of S. enterica subsp. enterica survival as a function of storage temperature was made. S. enterica subsp. enterica growth was inhibited during storage, and bacteria survived for an extensive period of time at high number (60 day at 5 °C, 26 day at 25 °C). The storage temperature significantly influenced the inactivation rate of Salmonella in raw milk ripened cheese and proceeded faster at 25 °C compared to remaining storage temperature. Obtained results suggest that contamination by Salmonella in raw milk cheese might result in residual risk.     

不同储存时间对普洱茶有机溶剂萃取物清除自由基活性的影响

本文以半数抑制浓度IC50的大小研究了十种茶叶（不同年份的生普和熟普）有机溶剂萃取物的体外抗氧化活性。结果表明,生茶有机溶剂萃取层清除DPPH自由基、羟基自由基、超氧自由基的IC50最小分别可达0.339 mg/mL、0.074 mg/mL、1.206 mg/mL,熟茶清除DPPH自由基、羟基自由基、超氧自由基的IC50最小分别可达0.554 mg/mL、0.164 mg/mL、1.927 mg/mL,生茶的抗氧化能力明显优于熟茶;各种普洱茶对自由基的清除能力主要取决于各萃取液中的多酚和黄酮含量。生茶乙酸乙酯层的茶多酚萃取量最多,在7.33-9.57%之间,正丁醇层的黄酮类物质的萃取量最多在7.08-10.99%之间,熟茶正丁醇层的茶多酚含量和黄酮类物质的含量在个有机溶剂层中均是最高的,分别在3.62-5.73%之间和4.75-5.83%之间,乙酸乙酯层和正丁醇层的萃取化合物具有较多的羟基,具有较强的清除自由基的能力;茶叶的储存时间对各萃取液的抗氧化能力无显著影响。