Quantitative analysis of flavan-3-ols in Spanish foodstuffs and beverages

An HPLC method, using detection after postcolumn derivatization with p-dimethylaminocynnamaldehyde (DMACA), was developed for the quantitative analysis of individual flavanols in food. This method was applied to flavanol determination in 56 different kinds of Spanish food products, including fruit, vegetables, legumes, beverages (cider, coffee, beer, tea, and wine), and chocolate. The determined compounds corresponded to the catechins and proanthocyanidin dimers and trimers usually present in food and, therefore, they were representative of the flavanols of low degree of polymerization consumed with the diet. The data generated could be used for calculation of the dietary intake of either individual or total flavanols, which would allow the further establishment of epidemiological correlations with the incidence of chronic diseases. Similar flavanol profiles were found in the different samples of a similar type of product, even though important variations could exist in the concentrations of total and individual flavanols among them. This was attributed to factors such as sample origin, stage of ripeness, post-harvesting conservation, and processing. Total flavanol contents varied from nondetectable in most of the vegetables to 184 mg/100 g found in a sample of broad bean. Substantial amounts were also found in some fruits, such as plum and apple, as well as in tea and red wine. Epicatechin was the most abundant flavanol, followed by catechin and procyanidin B2. In general, catechins were found in all the flavanol-containing products, but the presence of gallocatechins was only relevant in pomegranate, broad bean, lentil, grape, wine, beer, and tea, and most of the berries. Galloyled flavanols were only detected in strawberry, medlar, grape, and tea.     

Total phenolic content, antioxidant activity, and cross-cultural consumer rejection threshold in white and red wines functionally enhanced with catechin-rich extracts

White and red wines spiked with catechin-rich green tea extract and grape seed extract were assessed for phenolic content, antioxidant activity, and cross-cultural consumer rejection thresholds in relation to wine as a functional food. Health functionality is an important factor in functional foods, and spiking pure compounds or plant extracts is an effective method to increase or control functionality. The total phenolic content and antioxidant activity were measured in wines spiked to different extract concentrations, namely, control and 50, 100, 200, 400, and 800 mg/L, to confirm the dose-response curves in both white and red wines. Consumer rejection thresholds (CRTs) were established for spiked wines in a Korean and in an Australian population. Our results showed that the green tea extract and grape seed extract increased the antioxidant activity dose dependently, and the CRTs varied considerably between the Korean and the Australian groups, with Koreans preferring wines spiked with green tea extract and Australians showing a preference for wines spiked with grape seed extract. These results have implications for producing wine products that are enhanced in phenolic compounds and targeted to different cultural groups.     

不同种类茶叶冲泡前后中红外光谱变化探析

为建立一种快速无损评价茶叶/茶汤化学品质和色泽的方法,本试验选取7种代表性茶叶样本为材料,利用傅里叶变换中红外光谱(FTIR)技术,以开水冲泡方式制备茶汤冻干粉,对茶叶与茶汤共14个样本进行FTIR光谱采集分析。通过聚类分析、光谱曲线拟合、定性分析,并结合常规化学法检测茶多酚、茶多糖、可溶性糖、游离氨基酸、主要儿茶素类和生物碱含量。结果表明,不同茶叶光谱明显不同,由于茶叶加工工艺造成的脂类、儿茶素类、咖啡碱、游离氨基酸及叶绿素含量差异,利用1 800~1 170 cm^-1狭窄区域的光谱特征,可实现茶叶分类。茶叶与对应茶汤的光谱也存在明显差异,体现了茶叶在冲泡过程中的物质变化。主要特征峰区(位)为:2 925/2 855 cm^-1代表脂类饱和烃链-CH₂振动,1 700 cm^-1峰位对应儿茶素类和咖啡碱,1 534 cm^-1对应着叶绿素脱镁过程,1 551/1 556 cm^-1和1 500/1 497 cm^-1为叶绿素中亚甲基桥的伸缩振动,1 516/1 517 cm^-1为木质素中的苯环C=C伸缩振动。本研究为市场中茶叶、新型速溶茶粉样品化学品质检测提供了一种有效、快速的无损分析方法。     

Simultaneous determination of seven bioactive components in Oolong tea Camellia sinensis: quality control by chemical composition and HPLC fingerprints

A simple and reliable method of high-performance liquid chromatography (HPLC) was developed for the quality control of oolong tea (the dry leaves of Camellia sinensis ): the quality control included the HPLC fingerprint and the quantitative determination of seven bioactive compounds chemicals, namely, (-)-gallocatechin, (-)-epigallocatechin, (-)-epigallocatechin gallate, caffeine, (-)-epicatechin, gallocatechin gallate, and (-)-epicatechin gallate. The developed analyses of the chemicals excelled in quantifying the chemicals in oolong tea. The chemical fingerprint of oolong tea was established using the raw materials of three main production sites in China, that is, Fujian (southern and northern parts), Taiwan, and Guangdong. The fingerprints from different cultivated sources were analyzed by hierarchical cluster analysis, similarity analysis, principal component analysis (PCA), analysis of variance (ANOVA), and discriminant analysis. The results indicated that the combination of chromatographic fingerprint and quantification analysEs could be used for the quality assessment of oolong tea and its derived products.     

Validation of a mass spectrometry method to quantify oak ellagitannins in wine samples

Detection and individual quantification of oak wood ellagitannins in oak barrel aged red wine samples are difficult mainly due to their low levels and the similarity between their structures. In this work, a quantification method using mass spectrometry has been developed and validated to quantify wine ellagitannins after sample fractionation with a previously reported method. The use of an internal standard is a requirement to correct mass signal variability. (-)-Gallocatechin, among the different tested compounds, was the only one that proved to be a suitable internal standard making possible the accurate and individual quantification of the main oak wood ellagitannins. The developed methodology has been used to detect and quantify these ellagitannins in different Spanish commercial wines, proving its usefulness.     

Quantitative analysis of red wine tannins using Fourier-transform mid-infrared spectrometry

Tannin content and composition are critical quality components of red wines. No spectroscopic method assessing these phenols in wine has been described so far. We report here a new method using Fourier transform mid-infrared (FT-MIR) spectroscopy and chemometric techniques for the quantitative analysis of red wine tannins. Calibration models were developed using protein precipitation and phloroglucinolysis as analytical reference methods. After spectra preprocessing, six different predictive partial least-squares (PLS) models were evaluated, including the use of interval selection procedures such as iPLS and CSMWPLS. PLS regression with full-range (650-4000 cm(-1)), second derivative of the spectra and phloroglucinolysis as the reference method gave the most accurate determination for tannin concentration (RMSEC = 2.6%, RMSEP = 9.4%, r = 0.995). The prediction of the mean degree of polymerization (mDP) of the tannins also gave a reasonable prediction (RMSEC = 6.7%, RMSEP = 10.3%, r = 0.958). These results represent the first step in the development of a spectroscopic methodology for the quantification of several phenolic compounds that are critical for wine quality.     

Analysis of benzothiazole in Italian wines using headspace solid-phase microextraction and gas chromatography-mass spectrometry

Benzothiazoles are a part of the molecular structure of a large number of natural products, biocides, drugs, food flavors, and industrial chemicals. They also appear in the environment mainly as a result of their production and use as rubber vulcanization accelerators. A new headspace solid-phase microextraction (HS-SPME) method for analysis of benzothiazole (BTH) in wine is described. This method is fast, inexpensive, and does not require solvents. The detection limit of BTH in wine was 45 ppt with linearity up to 100 ppb. The quantification of BTH is performed by the standard additions method and does not require the use of an internal standard. We have analyzed 12 wines from different grape varieties grown in several regions, using SPME extraction and gas chromatography-mass spectrometry (GC-MS) detection. Under these experimental conditions, benzothiazole was found in all wines analyzed. Concentration levels in samples varied from 0.24 microg/L (Vermentino) to 1.09 microg/L (Franciacorta).     

Dispersive cleanup of acetonitrile extracts of tea samples by mixed multiwalled carbon nanotubes, primary secondary amine, and graphitized carbon black sorbents

A method for analysis of 37 pesticide residues in tea samples was developed and validated and was based on reversed-dispersive solid-phase extraction (r-DSPE) cleanup in acetonitrile solution, followed by liquid chromatography-electrospray tandem mass spectrometry determination. Green tea, oolong tea, and puer tea were selected as matrixes and represent the majority of tea types. Acetonitrile was used as the extraction solvent, with sodium chloride and magnesium sulfate enhancing partitioning of analytes into the organic phase. The extract was then cleaned up by r-DSPE using a mixture of multiwalled carbon nanotubes, primary secondary amine, and graphitized carbon black as sorbents to absorb interferences. Further optimization of sample preparation and determination allowed recoveries of between 70% and 111% for all 37 pesticides with relative standard deviations lower than 14% at two concentration levels of 10 and 100 μg kg(-1). Limits of quantification ranged from 5 to 20 μg kg(-1) for all pesticides. The developed method was successfully applied to the determination of pesticide residues in market tea samples.     

Rapid differentiation of tea products by surface desorption atmospheric pressure chemical ionization mass spectrometry

Protonated water molecules generated by an ambient corona discharge were directed to impact tea leaves for desorption/ionization at atmospheric pressure. Thus, a novel method based on surface desorption chemical ionization mass spectrometry (DAPCI-MS) has been developed for rapid analysis of tea products without any sample pretreatment. Under the optimized experimental conditions, DAPCI MS spectra of various tea samples are recorded rapidly, and the resulting mass spectra are chemical fingerprints that characterize the tea samples. On the basis of the mass spectral fingerprints, 40 tea samples including green tea, oolong tea, and jasmine tea were successfully differentiated by principal component analysis (PCA) of the mass spectral raw data. The PCA results were also validated with cluster analysis and supervised PCA analysis. The alteration of signal intensity caused by rough surfaces of tea leaves did not cause failure in the separation of the tea products. The experimental findings show that DAPCI-MS creates ions of both volatile and nonvolatile compounds in tea products at atmospheric pressure, providing a practical and convenient tool for high-throughput differentiation of tea products.     

Simultaneous analysis of free and conjugated estrogens, sulfonamides, and tetracyclines in runoff water and soils using solid-phase extraction and liquid chromatography-tandem mass spectrometry

The ability to monitor multiple analytes from various classes of compounds in a single analysis can increase throughput and reduce cost when compared to traditional methods of analyses. This method for analyzing free (parent estrogen) and conjugated estrogens (metabolites) along with sulfonamides and tetracyclines utilizes a high pH (10.4) mobile phase with an ammonium hydroxide buffer for both positive- and negative-mode electrospray ionization. A single-step sample preparation by solid-phase extraction (SPE) was used to isolate and concentrate all analytes simultaneously. The analytical method was developed and validated for recoveries at 3 concentration levels for water and soil and produced recoveries of 42-123% and 21-105% respectively. Method detection limits ranged from 0.3 to 1.0 ng/L for water samples and 0.01 to 0.1 ng/g for soils. The method quantification limit ranged from 0.9 to 3.3 ng/L for water samples and 0.06 to 0.7 ng/g for soils. The single-point standard addition calibration procedure was validated across a linear range of MQL to 100 ng/L with ≥82% accuracy against a matrix matched standard curve. Furthermore, sorption of tetracyclines onto glassware was investigated and minimized by 10% using nitric acid-rinsed glassware, while separation parameters were further optimized based on retention time and signal responses. This method has been used for the quantification of estrogens, tetracyclines, and sulfonamides in soil and runoff waters with multiple compounds detected simultaneously in a single analysis.     

Direct HPLC analysis of quercetin and trans-resveratrol in red wine,grape, and winemaking byproducts

A simple and fast reversed-phase HPLC method using diode array detection was developed and validated for the simultaneous determination of trans-resveratrol and quercetin in Sicilian red wine from the Nero d'Avola red grape variety. Investigation was also extended to the quantitative determination of resveratrol and quercetin in grape skins and winemaking byproducts obtained from the same cultivar. Samples were eluted using a C18 narrow-bore column under isocratic conditions in less than 20 min. Quantification of trans-resveratrol and quercetin in red wine was performed without any sample pretreatment, whereas the determination of these phenolic compounds in grape skins and wine pomage required a solvent extraction procedure. Linearity was demonstrated over the 0.39-12.5 and 0.45-57.6 microg/mL range for trans-resveratrol and quercetin, respectively. Detection limits in real samples were in the low ppm level (0.07 mg/L for trans-resveratrol and 0.12 mg/L for quercetin). The HPLC-UV/DAD method was applied for the routine analyses of red wine and grape skin and winemaking byproduct extracts to evaluate their trans-resveratrol and quercetin content. In particular, a very high content of quercetin was found in wine pomace, suggesting the use of this wine byproduct as a potential source of this health-promoting phenolic compound.     

Quantitative colorimetric assay for total protein applied to the red wine Pinot noir

A standard method for assaying protein in red wine is currently lacking. The method described here is based on protein precipitation followed by dye binding quantification. Improvements over existing approaches include minimal sample processing prior to protein precipitation with cold trichloroacetic acid/acetone and quantification based on absorbance relative to a commercially available standard representative of proteins likely to be found in wine, the yeast mannoprotein invertase. The precipitation method shortened preparation time relative to currently published methods and the mannoprotein standard yielded values comparable to those obtained by micro-Kjeldahl analysis. The assay was used to measure protein in 48 Pinot noir wines from 6 to 32 years old. The protein content of these wines was found to range from 50 to 102 mg/L with a mean value of 70 mg/L. The availability of a simple and relatively rapid procedure for assaying protein provides a practical tool to quantify a wine component that has been overlooked in routine analyses of red wines.     

Influence of prefermentative treatments to the major volatile compounds of Assyrtiko wines

A study of the volatile fraction of Assyrtiko wines, using gas chromatography coupled with olfactometry, was realized. Twenty-seven volatile compounds were identified as potent odorants, most of them originating from the fermentation process. Quantification of the major volatile compounds was realized developing a rapid analytical method based on fractionation of a 50 mL wine aliquot using C 18-reversed phase adsorbent. After elution of the volatile compounds with pentane-diethyl ether and concentration under nitrogen, the final wine extract was injected in a gas chromatography-flame ionization detection system. The method allows satisfactory determination of more than 15 volatile compounds of wine. The linearity of the method gave a typical r (2) between 0.990 and 0.999, while reproducibility ranged from 5.1 to 12.2% (as relative standard deviation) with 9.5% as the average. The method was applied to wines produced by Assyrtiko grapes (AOC Santorini), for two consecutive years, to compare the effect of skin contact prior to fermentation and the must clarification process. Direct press and skin contact wines were differentiated analytically; however, highly significant differences were not. Inversely, the differences found between direct press/clarified and nonclarified wines were significant. Wines produced by direct press and clarified must presented significantly higher levels of ethylic esters and fusel alcohol acetates but lower fusel alcohol levels, leading probably to more fruity wines. This difference, between clarified and nonclarified grape musts, was not significant in the case of the wines produced by skin contact of Assyrtiko berries. These findings were validated by preference sensory analysis tests.     

不同浸渍工艺对贵人香干白葡萄酒香气品质的影响

为提升贵人香干白葡萄酒的香气品质,以澄清汁发酵工艺(CQ)为对照,利用顶空固相微萃取结合气相色谱质谱联用技术(HS-SPME/GC-MS)检测浸渍工艺(JZ)和浸渍澄清工艺(JC)处理发酵酒样中的挥发性香气化合物。结果表明,浸渍处理可以明显增加葡萄酒中香气物质的含量。与CQ相比,JZ和JC对发酵酒样中品种香气物质的含量影响显著(P<0.05),萜烯类化合物含量分别增加1.86、1.01倍,且JC酒样中醇类、酯类香气物质含量最高,分别为4 799.38 μg·L^-1和5 013.28 μg·L^-1。主成分分析结合感官评价结果表明,JZ和JC酒样的花香特征相似,且JC酒样的热带水果香气以及醇香特征更为明显;与CQ相比,JC发酵酒样香气优雅、纯正,口感平衡。综合分析,浸渍澄清工艺(JC)可有效提升甘肃河西产区贵人香干白葡萄酒的香气品质。本研究结果可为干白葡萄酒增香酿造提供技术支持。     

Direct liquid chromatography method for the simultaneous quantification of hydroxytyrosol and tyrosol in red wines

A validated HPLC method with fluorescence detection for the simultaneous quantification of hydroxytyrosol and tyrosol in red wines is described. Detection conditions for both compounds were optimized (excitation at 279 and 278 and emission at 631 and 598 nm for hydroxytyrosol and tyrosol, respectively). The validation of the analytical method was based on selectivity, linearity, robustness, detection and quantification limits, repeatability, and recovery. The detection and quantification limits in red wines were set at 0.023 and 0.076 mg L(-1) for hydroxytyrosol and at 0.007 and 0.024 mg L(-1) for tyrosol determination, respectively. Precision values, both within-day and between-day (n = 5), remained below 3% for both compounds. In addition, a fractional factorial experimental design was developed to analyze the influence of six different conditions on analysis. The final optimized HPLC-fluorescence method allowed the analysis of 30 nonpretreated Spanish red wines to evaluate their hydroxytyrosol and tyrosol contents.     

Rapid isolation of red wine polymeric polyphenols by solid-phase extraction

A rapid technique for the isolation of polymeric polyphenols from red wine has been developed and validated. A copolymer reversed-phase SPE cartridge was utilized in conjunction with predominantly organic eluents to provide three phenolic fractions from red wine without the need for sample pretreatment. The first fraction contained the bulk of the monomeric and oligomeric phenolic material, while the second and third fractions contained the polymeric polyphenolic compounds, as determined by HPLC analysis. The two polymeric polyphenolic fractions differed in their solubility and extent of pigmentation, and the differences appeared to be related to wine age. This method contrasted with other available fractionation techniques because the interfering, nonpolymeric material can be removed in a single wash fraction, while the polymeric material is separated into two distinct fractions based on their diverse physicochemical properties. It is anticipated that the rapid access to discrete polymeric fractions afforded by this method will be of benefit in furthering the understanding of red wine polymeric polyphenols.     

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.     

1H NMR based metabolic profiling in the evaluation of Japanese green tea quality

Classification of tea quality is now mainly performed according to the sensory results by professional tea tasters. However, this evaluation method is inconsistent in differentiating their qualities. A combination of a (1)H NMR technique and a multivariate analysis was introduced to the quality evaluation of green tea by means of a metabolomic technique. A broad range of metabolites were detected by (1)H NMR spectrometry. The principal component analysis (PCA) was used to reduce the complexity of the (1)H NMR spectra data set and provided the quality discrimination result. It offered an extensive clue for classification and quality assessment without any prepurification method. A set of green teas from a Japanese tea contest were analyzed by (1)H NMR to classify the quality with respect to that judged by tea tasters and to conceive a quality prediction model. Metabolic profiling and fingerprinting of (1)H NMR spectra of green teas with different quality were studied. PCA showed a separation between the high- and the low-quality green teas. The taste marker compounds contributing to the discrimination of tea quality were identified. Reliable prediction models were obtained by the partial least-squares projection to latent structure (PLS) analysis together with a preprocessing filter of both orthogonal signal correction (OSC) and a combination between OSC and wavelet transform algorithms.     

Survey on the presence of butyltin compounds in chinese alcoholic beverages,determined by using headspace solid-phase microextraction coupled with gas chromatography-flame photometric detection

The use of butyltin compounds in some food packaging leads to the contamination of liquid food and may result in subsequent adverse effects on people's health through the food chain. A survey of butyltin compounds in Chinese alcoholic beverages purchased from retail markets was carried out by using solid-phase microextraction (SPME) followed by gas chromatography coupled with flame photometric detection. Forty-four samples including wine, liquor, and champagne were studied. The levels of monobutyltin and dibutyltin ranged from <0.016 to 5.687 and from <0.0022 to 33.257 microg of Sn/L, respectively. Low levels of tributyltin were detected. The presence of dibutyltin in wine samples was further confirmed by GC-MS. The result indicated that dry wines generally contained more dibutyltin than sweet wines. The concentrations of butyltin compounds in liquor samples were lower than those in wine samples.     

Plant foods and herbal sources of resveratrol

Stilbenes, in particular trans-resveratrol and its glucoside, are widely reported to be beneficial to health, having been shown to possess antioxidative, anticarcinogenic, and antitumor properties. Major dietary sources include grapes, wine, peanuts, and soy; however, they can also be introduced into the diet through Itadori tea, which has long been used in Japan and China as a traditional herbal remedy for heart disease and strokes. Analysis of grapes, peanuts, and Itadori tea shows that they contain mainly trans-resveratrol glucoside. In contrast, red wines are primarily a source of the aglycones cis- and trans-resveratrol. While peanuts and grapes contain low levels of the stilbenes, Itadori tea and red wine both supply relatively high concentrations of resveratrol. For people who do not consume alcohol, Itadori tea may be a suitable substitute for red wine. However, further study on the potential biological effects of other endogenous compounds in Itadori tea is required and there is also a need for more information on the absorption and in vivo biomedical actions of free and conjugated resveratrol.