A fast and accurate method for controlling the correct labeling of products containing buffalo meat using High Resolution Melting (HRM) analysis

The substitution of high priced meat with low cost ones and the fraudulent labeling of meat products make the identification and traceability of meat species and their processed products in the food chain important. A polymerase chain reaction followed by a High Resolution Melting (HRM) analysis was developed for species specific detection of buffalo; it was applied in six commercial meat products. A pair of specific 12S and universal 18S rRNA primers were employed and yielded DNA fragments of 220 bp and 77 bp, respectively. All tested products were found to contain buffalo meat and presented melting curves with at least two visible inflection points derived from the amplicons of the 12S specific and 18S universal primers. The presence of buffalo meat in meat products and the adulteration of buffalo products with unknown species were established down to a level of 0.1%. HRM was proven to be a fast and accurate technique for authentication testing of meat products.     

Barcode High Resolution Melting (Bar-HRM) analysis for detection and quantification of PDO “Fava Santorinis” (Lathyrus clymenum) adulterants

Legumes considered as one of the most important crops worldwide. Due to high price as a PDO product, commercial products of “Fava Santorinis” are often subjected to adulterations from other legume products coming from other Lathyrus or Vicia and Pisum species. Using plant DNA barcoding regions (trnL and rpoC) coupled with High Resolution Melting (Bar-HRM) we have developed a method allowing us to detect and authenticate PDO “Fava Santorinis”. Bar-HRM proved to be a very sensitive tool able to genotype Lathyrus and its closed relative species and to detect admixtures, being sensitive enough to as low as 1:100 of non-“Fava Santorinis” in “Fava Santorinis” commercial products. In conclusion, Bar-HRM analysis can be a faster, with higher resolution and cost effectiveness alternative method to authenticate PDO “Fava Santorinis” and to quantitatively detect adulterations in “Fava Santorinis” with other relative commercial “Fava” food products.     

Essential oil composition of commercial black tea (Camellia sinensis)

The essential oil components of different tea brands were investigated by gas chromatography. The oil yields of dried tea samples were ranged from 0.09% to 0.63%. Twenty-five compounds from Supreme and Lipton Yellow Label tea brands representing 98.0% and 88.0% of the Camellia sinensis oil were identified, respectively. The main ones were β-pinene (51.2%) and α-pinene (30.2%). Nineteen components from Tapal tea brand representing 76.7% of the C. sinensis oil were determined with high contents of muurol-5-en-4-a-ol (10.5%) and muurol-5-en-4-b-ol (31.3%). Fifteen components from Deer and Diana tea brands were identified, accounting for 83.3% and 78.2% of the oil containing α-cadinol and β-pinene. Seventeen components from non-branded teas were determined with high contents of muurol-5-en-4-a-ol and muurol-5-en-4-b-ol. Twenty-one compounds from non-branded Bangladeshi Shezan and Indian teas were also identified. All oils consisted of monoterpenic hydrocarbons, oxygenated monoterpenes and sesquiterpenes.     

Adulterations in Basmati rice detected quantitatively by combined use of microsatellite and fragrance typing with High Resolution Melting (HRM) analysis

The aim of this work was to setup a DNA based method coupled with High Resolution Melting (HRM) analysis for rice products traceability using five different microsatellite markers to genotyping Basmati and non-Basmati varieties. We also exploit the obtained information to detect the presence of Basmati varieties in commercial rice products. Additionally we used the 8 bp deletion in badh2 gene in combination with HRM to both DNA-typing of the Basmati and non-Basmati varieties and to quantitate accurately adulteration of Basmati rice products with non-Basmati rice products. HRM proved to be a very sensitive tool to genotype rice varieties and detect admixtures as well as able to detect a ratio of 1:100 of non-fragrance in fragrance rice. In conclusion HRM analysis can be a higher resolution, cost effective, alternative method compared to other techniques that could be extended to quantify adulterations in rice varieties and commercial rice food products.     

Uptake of fluoride by tea plant (Camellia sinensis L) and the impact of aluminium

Tea plant (Camellia sinensis L) accumulates large amounts of fluoride in mature leaves from soils of normal fluoride availabilities, but the properties of fluoride absorption by this plant species are not well understood. The present study examined the characteristic of fluoride uptake by tea plants and the impact of Al with solution and soil experiments. The results showed that the fluoride taken up by tea plants was largely and readily transported, in particular to the leaves. The fluoride in leaves increased linearly with fluoride concentrations in the uptake solution (R² = 0.75, P < 0.001) or in the soil (R² = 0.88, P < 0.001), whereas those in root and stem were only marginally influenced. Including Al in solution or adding Al to the soil apparently increased the uptake and translocation of fluoride to the leaves. The concentrations of fluoride in the leaves were significantly (P < 0.05) increased by 19.1% or 37.7% when 18.5 µmol l^?1 Al or 74.1 µmol l^?1 Al respectively was included in the uptake solution, compared with the control without Al during an uptake period of 22 h. Similarly, Al application (100 mg kg^?1) to soil led to significantly higher fluoride concentrations in mature leaves and new shoots (one bud with three leaves). By contrast, the concentrations of Al in leaves in solution and soil experiments were not affected by fluoride and Al treatments. Nevertheless, higher Al concentrations after Al and fluoride additions were observed in the new shoots in soil experiments. Copyright © 2003 Society of Chemical Industry     

Microsatellite genotyping with HRM (High Resolution Melting) analysis for identification of the PGI common bean variety Plake Megalosperma Prespon

Legumes and common bean (Phaseolus vulgaris L.), in particular, are important crops worldwide, consumed either as dried seeds or fresh fruits. Correct identification of common bean varieties is important, in order to ensure food quality, safety and authenticity for consumers. Recently, DNA based methods, including molecular markers like microsatellites (SSR), have been developed for plant species or variety identification genotyping and for identification of their ingredients in the final food products. Here, we have applied High Resolution Melting (HRM) analysis coupled with four microsatellite markers to facilitate the identification of protected geographic indication (PGI) common bean variety ‘Plake Megalosperma Prespon’ (‘PMP’). The four microsatellite loci used were informative and were used to generate a unique melting curve profile of microsatellites for each variety tested. These microsatellite markers enabled the distinction and identification of the PGI (common bean variety ‘PMP’). Hence, this assay provided a flexible, cost-effective and closed-tube microsatellite genotyping method, well suited to varietal identification and authentication analysis in common beans.     

单核苷酸多态性在茶树中应用的研究进展

茶树原产于中国西南地区,是一种重要的经济作物。茶叶作为中国的传统饮品,是世界上三大无酒精饮料之一。长期的自然演化及自交不亲和使得茶树的种质资源极其丰富。单核苷酸多态性(single nucleotide polymorphism,SNP)是基因组水平上单核苷酸突变引起的基因序列多样性,是继限制性酶切片段多态性(restriction fragment length polymorphism,RFLP)和简单序列重复(simple sequence repeat,SSR)的第3代分子标记,广泛存在于生物基因组中。SNP分子标记作为最重要的分子标记技术对遗传学研究有极其重要的作用。本文介绍了SNP分子标记的特征,综述了SNP在茶树中的开发检测方法,研究应用于分子标记、基因定位、关联分析等,旨在为理解和研究SNP分子标记在茶树中的研究应用提供参考。     

Understanding the biosyntheses and stress response mechanisms of aroma compounds in tea (Camellia sinensis) to safely and effectively improve tea aroma

Abstract

Metabolite formation is a biochemical and physiological feature of plants developed as an environmental response during the evolutionary process. These metabolites help defend plants against environmental stresses, but are also important quality components in crops. Utilizing the stress response to improve natural quality components in plants has attracted increasing research interest. Tea, which is processed by the tender shoots or leaves of tea plant (Camellia sinensis (L.) O. Kuntze), is the second most popular beverage worldwide after water. Aroma is an important factor affecting tea character and quality. The defense responses of tea leaves against various stresses during preharvest (tea growth process) and postharvest (tea manufacturing) processing can result in aroma formation. Herein, we summarize recent investigations into the biosyntheses of several characteristic aroma compounds prevalent in teas and derived from volatile fatty acid derivatives, terpenes, and phenylpropanoids/benzenoids. Several key aroma synthetic genes from tea leaves have been isolated, cloned, sequenced, and functionally characterized. Biotic stress (such as tea green leafhopper attack) and abiotic stress (such as light, temperature, and wounding) could enhance the expression of aroma synthetic genes, resulting in the abundant accumulation of characteristic aroma compounds in tea leaves. Understanding the specific relationships between characteristic aroma compounds and stresses is key to improving tea quality safely and effectively.

     

Leaching and bioavailability of aluminium, copper and manganese from tea (Camellia sinensis)

To assess human exposure to Al, Cu and Mn from tea drinking, eight tea samples were analysed for their ‘total’ and soluble elemental content. From this data, the percentage of ‘available’ element to the human system was calculated. Results for the samples showed that, compared to background levels for plants, concentrations of Al were highly elevated. The solubility of Al, Cu and Mn in infusions at 2, 5 and 10 min with boiling water showed that the solubility in the first infusion was the highest followed by the second and the third infusions in decreasing order. Calculation of percentage ‘available’ Al, Cu and Mn to the human system showed that tea is a rich source of dietary Mn, contributes towards our dietary copper intake, and is of no cause for concern in terms of Al toxicity in healthy individuals.     

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     

Multielement analysis of Chinese tea (Camellia sinensis) by total-reflection X-ray fluorescence

 Total-reflection X–ray fluorescence (TXRF) was used for the simultaneous determination of 15 elements in tea samples which were produced either by acid digestion or acidified infusion of tea leaves (Camellia sinensis). The accuracy and precision of the method were checked by its application to a certified reference material (GBW 08505 : tea). A variety of 39 tea samples of different kinds and/or qualities produced in different regions of China were analysed. The range and mean of the concentrations of elements in the tea leaves (0.1–30.000 μg g^–1) and their solubility in infusions (0.5–85%) were determined and the influence of the origin, type and quality of the tea samples was studied. In some tea leaves produced in a Se-rich region, the content of Se was found to be very high (up to 7.5 μg g^–1), in contrast to a concentration of only about 0.1 μg g^–1 Se in most of the tea leaves examined. Received: 27 January 1998     

高分辨率熔解曲线分析法检测食源性副溶血性弧菌

为建立一种快速鉴定副溶血性弧茵的HRM（高分辨率熔解曲线）real-timePCR法,以toxR为靶基因,结合特异性引物,通过优化反应体系及条件,进行特异性、敏感性及重复性评价,并初步应用于90份送检的鲜活海产品样本的检测。特异性试睑表明,该方法能选择性检测副溶血弧菌,Tm值为76．64±0．57℃;而与创伤弧菌、霍乱弧茵、金黄色葡萄球菌等多种海产品中常见的食源性病原菌没有交叉反应。灵敏度试验表明,该方法最少可检测toxR重组质粒的浓度为3．50×102copies／mL。重复性试验表明,同一样品于试验内及试验问的平均Tm值分别为76．53±0．35℃和76．74±0．52℃,变异系数分别为0．56±0．42％和1．11±0．73％。对90份鲜活海产品样本的检测证实该法可使阳性检出率从国标法的14．44%高至18．89％。本研究所建立的副溶血性弧菌HRMreal-ILrnePCR法具有特异性好、灵敏度高、重复性好的特点,能应用于食品样本的检测,具有很好的研究价值和应用前景。     

Rapid detection and quantification of milk adulteration using infrared microspectroscopy and chemometrics analysis

The application of attenuated total reflectance mid-infrared microspectroscopy (MIR-microspectroscopy) was evaluated as a rapid method for detection and quantification of milk adulteration. Milk samples were purchased from local grocery stores (Columbus, OH, USA) and spiked at different concentrations of whey, hydrogen peroxide, synthetic urine, urea and synthetic milk. Samples were place on a 192-well microarray slide, air-dried and spectra were collected by using MIR-microspectroscopy. Pattern recognition analysis by Soft Independent Modeling of Class Analogy (SIMCA) showed tight and well-separated clusters allowing discrimination of control samples from adulterated milk. Partial Least Squares Regression (PLSR) showed standard error of prediction (SEP) ∼2.33, 0.06, 0.41, 0.30 and 0.014 g/L for estimation of levels of adulteration with whey, synthetic milk, synthetic urine, urea and hydrogen peroxide, respectively. Results showed that MIR-microspectroscopy can provide an alternative methodology to the dairy industry for screening potential fraudulent practice for economic adulteration of cow’s milk.     

Influence of some bioregulators on quality traits of pruned tea (Camellia sinensis (L) O Kuntze)

During investigation, it was observed that all the applied bioregulators, namely, jibika, IAA, cycocel, thiourea, methanol, succinic acid and sucrose, have a significant effect on quality parameters such as polyphenol oxidase (PPO) activity, caffeine, crude protein, starch, nitrogen, carotenoid and ascorbic acid (vitamin C) content over the control. However, maximum PPO activity was observed with cycocel followed by succinic acid, jibika, thiourea and sucrose during the first phase of spray while at the second phase PPO was maximal in sucrose-treated bushes followed by cycocel, jibika, thiourea and succinic acid. The caffeine content was found to be maximal in methanol followed by cycocel, IAA, thiourea and jibika as compared to the control during the first phase of spray. Similarly, nitrogen content increased due to methanol application at both the phases. Starch and carotenoid content were significantly influenced by jibika treatment. Likewise, ascorbic acid content was highest in sucrose-treated bushes as compared to other treatments. © 1998 SCI.     

食醋中勾兑工业合成醋酸的调查分析

目的了解杭州地区市售食醋勾兑醋酸的现状,为今后质量监督执法和评价提供科学依据。方法对杭州地区流通领域的食醋进行了抽样检测分析,采用相关强制性国家标准进行判定。结果本次对杭州地区流通领域的61个食醋样品进行了勾兑合成醋酸检测分析,其中酿造食醋51个样品,配制食醋10个样品,根据GB1903—2008《食品添加剂冰乙酸(冰醋酸)》标准酿造醋酸比率(天然度)%≥95%判定,酿造食醋合格率为96.08%,配制食醋合格率为70.00%,配制食醋的掺假现象比酿造食醋严重。结论今后应进一步加强食醋勾兑合成醋酸的风险监测力度,确保食醋质量安全。     

Differentiation of eight tea (Camellia sinensis) cultivars in China by elemental fingerprint of their leaves

BACKGROUND: Tea is an infusion made from dried leaves of tea (Camellia sinensis) and can be a good dietary source of essential trace metals for humans. Therefore, it is necessary to consider variations in element content of tea leaves among tea cultivars. Thus, elemental fingerprint techniques, based on elemental contents (Al, B, Ca, Cd, Co, Cr, Cu, Fe, K, Mg, Mn, Na, P, Pb, and Zn) determined by inductively coupled plasma atomic emission spectrometry (ICP‐AES) and multivariate statistical analysis, have been used to differentiate eight tea cultivars. RESULTS: The ranges of element concentrations in leaves of the eight cultivars were in good agreement with those obtained in previous studies and the level of most elements in tea leaves was significantly different among cultivars. The classifications of eight tea cultivars were 100% accurate in total by principal component analysis (PCA), hierarchical cluster analysis (HCA), linear discriminant analysis (LDA), and back‐propagation neural network (BPNN) analysis. CONCLUSION: Each cultivar presented a distinctive element fingerprint and the elements in tea leaves can be significant predictors in differentiating tea cultivars. Copyright © 2009 Society of Chemical Industry