Assessment of anthocyanins in grape (Vitis vinifera L.) berries using a noninvasive chlorophyll fluorescence method

Anthocyanins (Anths) in grape (Vitis vinifera L.) berries harvested at véraison from Pinot Noir and Pinot Meunier cultivars were assessed nondestructively by measuring chlorophyll fluorescence (ChlF) excitation spectra. With increasing Anth content, less excitation light was transmitted to the deeper Chl layers, and thus the ChlF signal decreased proportionally. By applying Beer-Lambert's law, the logarithm of the ratio between the fluorescence excitation spectra (log FER) from a green and a red berry gave the in vivo absorption spectrum of Anths, which peaked at about 540 nm. Absolute quantitative nondestructive determination of Anths for each berry was obtained by the log FER calculated for two excitation wavelengths, 540 and 635 nm (absorbed and not-absorbed by Anths, respectively) of ChlF at 685 nm. Over a range of skin colors going from green to purple, the relationship between the log [ChlF(635)/ChlF(540)] and the Anth concentration of berry extracts was fairly well fitted (r 2 = 0.92) using a power function. Reflectance spectra on the same berry samples were also measured, and Anth reflectance indices, which were originally developed for apples and table grapes, were derived. The log FER Anth index was superior to the reflectance-ratio-based index, but was as good as the color index for red grapes (CIRG) calculated from the whole visible reflectance spectrum. The proposed log FER method, applied by means of suitable portable devices, may represent a new, rapid, and noninvasive tool for the assessment of grape phenolic maturity in vineyards.     

Characterization of odor-active compounds in extracts obtained by simultaneous extraction/distillation from moroccan black olives

"Greek-style" Moroccan black table olives were screened for potent odorants by GC/olfactometry/aroma extract dilution analysis of representative Likens-Nickerson extracts and compared with "Spanish-style" green fruits. ( Z)-3-Hexenal, ( E, E)-2,4-decadienal, ( E, Z)-2,4-decadienal, guaiacol, and methional were found in both green and black olives, but with significant differences in concentration according to the fruit ripening degree (the first was lower and the last two were higher in black fruits). Specific compounds not previously detected in green olives (gamma-deca- and dodecalactones, delta-decalactone, and 2-methyl-3-furanthiol) proved to be, with methional, the strongest odors in black olive extracts. These extracts were also distinguishable from green olive extracts by the presence of new sulfur compounds and fewer terpenes.     

Separation of olive proteins combining a simple extraction method and a selective capillary electrophoresis (CE) approach: application to raw and table olive samples

A simple extraction method was developed to extract proteins from olive samples based on chloroform/methanol extraction followed by a protein precipitation with cold acetone. Then, a capillary electrophoresis (CE) method was carried out using an acid buffer (1 M formic acid at pH 2) to ensure a positive net charge for proteins and a neutral charge for potential interferents as polyphenols. The method developed was applied to raw and table olive samples. Interestingly, raw olive samples showed differences in protein profiles depending upon the botanical variety of olives and their geographical region. Protein profiles obtained for table olives also showed differences according to the sample treatment. Thus, a signal reduction in the electropherograms obtained for black olives was observed in comparison to those achieved for treated green olives. In this work, the use of protein profiles was demonstrated to be a powerful tool for studying variations among olive samples.     

Simultaneous fluorometric determination of chlorophylls a and B and pheophytins a and B in olive oil by partial least-squares calibration

The resolution of quaternary mixtures of chlorophylls a and b and pheophytins a and b has been accomplished by partial least-squares (PLS) multivariate calibration, applied to the fluorescence signals of these pigments. The total luminescence information of the compounds has been used to optimize the spectral data set to perform the calibration. After preliminary studies, a method is described in acetone media, to avoid emulsions with the olive oil samples. Different scanning paths have been selected for each method. For the simultaneous determination of the pigments in olive oil samples, a comparative study of the results found by using excitation, emission, and synchronous spectral data, as analytical signal, was performed. The excitation spectra were selected as the better analytical signals for the determination of the pigments in olive oil samples. The optimum wavelength range to record the excitation spectra (lambda(em) = 662 nm) was selected to minimize the contribution of pheophytin a and to maximize the contribution of the other pigments, which are the minor constituents in olive oil. Determination of these pigments in olive oil samples was effected from the excitation spectra of dissolutions o suitable aliquots in acetone. Recovery values from olive oil, spiked with chlorophylls a and b and pheophytins a and b, were in the ranges of 70-112, 71-111, 76-105, and 82-109%, respectively.     

Discrimination of olives according to fruit quality using Fourier transform Raman spectroscopy and pattern recognition techniques

Fourier transform Raman spectroscopy combined with pattern recognition has been used to discriminate olives of different qualities. They included samples of sound olives, olives with frostbite, olives that have been collected from the ground, fermented olives, and olive samples with diseases. Milled olives were measured in a dedicated sample cup, which was rotated during spectrum acquisition. A preliminary study of the data set structure was performed using hierarchical cluster analysis and principal component analysis. Two supervised pattern recognition techniques, K-nearest neighbors and soft independent modeling of class analogy (SIMCA), were tested using a "leave-a-fourth-out" cross-validation procedure. SIMCA provided the best results, with prediction abilities of 95% for sound, 93% for frostbite, 96% for ground, and 92% for fermented olives. The olive samples with diseases (too few to define a class) were included in the validation and recognized as not belonging to any class. None of the damaged olive samples was wrongly predicted to the class of sound olives. With this approach a selection of sound olives for the production of high-quality virgin olive oil can be achieved.     

The activity of healthy olive microbiota during virgin olive oil extraction influences oil chemical composition

The activity of olive microbiota during the oil extraction process could be a critical point for virgin olive oil quality. With the aim to evaluate the role of microbiological activity during the virgin olive oil extraction process, just before oil extraction freshly collected healthy olive fruits were immersed in contaminated water from an olive mill washing tank. The oils extracted were then compared with control samples from the same batch of hand-picked olives. The presence of lactic and enteric bacteria, fungi and Pseudomonas on the surface of olives was proved to be much higher in washed than in control olives, with increments in cfu/g between 2 and 3 orders of magnitude. The biogenesis of volatile compounds and the extraction of olive polyphenols and pigments were significantly influenced by the microbiological profile of olives even without any previous storage. In most cases the effect of olive microbiota on oil characteristics was greater than the effect exerted by malaxation time and temperature. Oils from microbiologically contaminated olives showed lower amounts of C5 volatiles and higher levels of C6 volatiles from the lipoxygenase pathway and some fermentation products. On the other hand, a decrease of chlorophylls, pheophytins, xanthophylls and the ratio chlorophyll/pheophytin was observed in these oils. Likewise, the microbiological activity during oil extraction led to significantly lower amounts of polyphenols, in particular of oleuropein derivatives. These differences in olive oil chemical composition were reflected in oil sensory characteristics by the decrease of the green and bitter attributes and by the modification of the oil color chromatic ordinates.     

Screening for key odorants in Moroccan green olives by gas chromatography-olfactometry/aroma extract dilution analysis

"Spanish style" Moroccan green table olives were screened for potent odorants by gas chromatography-olfactometry/aroma extraction dilution analysis of a representative Likens-Nickerson extract. (Z)-3-Hexenal [flavor dilution factor (FD) = 256], (E,E)-2,4-decadienal (FD = 128), and (E,Z)-2,4-decadienal (FD = 64) were revealed to confer green and coriander/paraffin oil odors to both fruit and oil extracts, whereas guaiacol (FD = 128) imparted a bad olive, phenolic note. Methional (3-methylthiopropionaldehyde, FD = 128) and several terpenes (FD 相似文献   

Cluster analysis applied to the exploratory analysis of commercial spanish olive oils by means of excitation-emission fluorescence spectroscopy

Olive oil fluorescence is related to oil composition. Here it is shown that the natural clustering of different types of commercial Spanish olive oils depends on their fluorescence excitation-emission matrices (EEMs). Fifty-six commercial samples of olive oil (29 virgin olive oils, 20 pure olive oils, and 7 olive-pomace oils) were used. The clustering method was hierarchical agglomerative clustering using the Euclidean distance as a similarity measure and the average linkage. Two spectral ranges were considered (which either contained the fluorescence peak of the chlorophylls or did not), and various methods for preprocessing the fluorescence spectra were compared. The oils were clearly distinguished using the unfolded EEMs measured between lambda(ex) = 300-400 nm and lambda(em) = 400-600 nm. The optimal preprocessing was normalization of the unfolded spectra followed by column autoscaling. Also shown are the advantages of using second-order data (EEMs) instead of first-order data (a single fluorescence spectrum) for each sample.     

Influence of harvesting method and washing on the presence of pesticide residues in olives and olive oil

The influence of the preliminary steps in olive oil production (harvesting and washing) on pesticide residues in olives and olive oil has been investigated. Analyses were performed by GC-MS/MS and revealed that endosulfan sulfate and two herbicides (diuron and terbuthylazine) were the most frequently found residues in olives and olive oil. The harvesting method has a decisive influence on herbicide concentrations found in olives. Thus, 16 and 48% of the olive samples harvested on the ground after falling from the tree presented concentrations higher than the maximum residue limit (MRL) for diuron and terbuthylazine, respectively. In olives harvested directly from the tree, diuron was not found at concentrations higher than MRL and terbuthylazine was found in only 10% of the samples. The washing step performed routinely in olive mills was effective in removing the superficial contamination by herbicides present in olives harvested on the ground. Nevertheless, even after washing, the olive oil obtained from ground olives showed herbicide residue concentrations higher than those obtained from tree olives.     

Origin of French virgin olive oil registered designation of origins predicted by chemometric analysis of synchronous excitation-emission fluorescence spectra

The authentication of virgin olive oil samples requires usually the use of sophisticated and very expensive analytical techniques, so there is a need for fast and inexpensive analytical techniques for use in a quality control methodology. Virgin olive oils present an intense fluorescence spectra. Synchronous excitation-emission fluorescence spectroscopy (SEEFS) was assessed for origin determination of virgin olive oil samples from five French registered designation of origins (RDOs) (Nyons, Vallée des Baux, Aix-en-Provence, Haute-Provence, and Nice). The spectra present bands between 600 and 700 nm in emission due to chlorophylls a and b and pheophytins a and b. The bands between 275 and 400 nm in emission were attributed to alpha-, beta-, and gamma-tocopherols and to phenolic compounds, which characterize the virgin olive oils compared to other edible oils. The chemometric treatment (PLS1) of synchronous excitation-emission fluorescence spectra allows one to determine the origin of the oils from five French RDOs (Baux, Aix, Haute-Provence, Nice, and Nyons). Results were quite satisfactory, despite the similarity between two denominations of origin (Baux and Aix) that are composed by some common cultivars (Aglandau and Salonenque). The interpretation of the regression coefficients shows that RDOs are correlated to chlorophylls, pheophytins, tocopherols, and phenols compounds, which are different for each origin. SEEFS is part of a global analytic methodology that associates spectroscopic and chromatographic techniques. This approach can be used for traceability and vindicates the RDOs.     

Pigment-lipoprotein complexes in table olives (Cv. Gordal) with green staining alteration

In table olives showing the green staining alteration, extracts of pigment-lipoprotein complexes were obtained from the zone altered and the rest of the fruit. In the altered zone of the olive, the surrounding components of pigments were affected, with the degeneration of proteins and phospholipids forming the pigment-lipoprotein complexes. There was also less interaction between the pigments and the membrane lipids. These results suggested a greater loss of cell integrity in the green-stained zone of the fruit, allowing the migration and local accumulation of Cu-metallochlorophyll complexes, macroscopically visible as the form of green staining alteration.     

Effects of olive fruit quality and oil storage practices on the diacylglycerol content of virgin olive oils

The evolution of 1,3- and 1,2-isomers of diacylglycerols (DGs) in olive oils obtained from healthy olives and the influence of the olive quality was studied. Healthy olive fruits yielded oils containing almost exclusively 1,2-isomers whereas altered olives produced oils with significant amounts of 1,3-isomers. Virgin olive oils obtained from various olive cultivars and stored at different temperatures showed triacylglycerol hydrolysis and diacylglycerol isomerization depending on the acidity and temperature. The results indicated that the relationship between acidity and total diacylglycerol content has scarce utility for detecting mild refined oil in virgin olive oil. On the other hand, the 1,3-/1,2-DG isomers ratio is useful for assessing the genuineness of virgin olive oils with low acidities during the early stages of storage.     

Fluorescence of vegetable oils: olive oils

Fluorescence spectra of undiluted extra virgin olive oil obtained with the traditional setup (right-angle fluorescence) show considerable artifacts and deformations due to self-absorption phenomena, even when the spectra are corrected for inner filter effects. On the other side, front-face fluorescence spectra are much less affected by self-absorption. Front-face fluorescence of native olive oil reveals the presence of different fluorophores and can provide information about their amount. From the intense emission at ca. 315-330 nm, it is possible to detect fluorescent polyphenols and pherols and to evaluate their overall content. Low-intensity emission bands at 350-600 nm are correlated to vitamins and other important molecules. Among them, the fluorescence of the riboflavin fluorophore can be used to evaluate its concentration. The intense emission of chlorophyll derivatives, measured in the 640-800 nm spectral region, can provide information on their concentration.     

Fluorometric determination of nitrite in cured meats.

An indirect fluorometric method for determining sodium nitrite in meat products is presented. The extracted sodium nitrite is consumed in a diazotization reaction with a measured excess of sulfanilic acid. Fluorescamine, which acts selectively with primary amines such as sulfanilic acid, is a fluorogenic reagent for the excess amine. The amine consumed, calculated by difference from the total originally present, is directly related to the sodium nitrite content of the sample. Interferences from amino acids and soluble proteins in the meat extract are eliminated by judicious use of a secondary peak in the fluorescence spectra (436 nm excitation, 495 nm fluorescence) combined with measurement at low pH (3.30). The recoveries of sodium nitrite ranged from 83.2 to 99.6% with an average of 93.4 and a standard deviation of +/- 5.28% for 11 determinations.     

基于特征荧光信号的去囊衣带芯橘瓣分选

为实现去囊衣带芯橘瓣的机器视觉分选,给去囊衣带芯橘瓣的机器识别提供直接、准确的判别信号,采用荧光分光光度计对50颗去囊衣带芯橘瓣的橘芯和砂囊分别进行三维荧光光谱扫描,通过对橘芯、砂囊平均三维荧光光谱分析,确定了橘芯相对砂囊的特征荧光信号,据此对带芯橘瓣荧光图像识别的可行性进行了验证。检测发现,在370~390 nm紫外光激发下,橘芯和砂囊在440 nm处的荧光强度差异较大;橘芯与砂囊在370 nm紫外光激发下,440 nm处荧光强度分布的箱线图表明两者荧光强度分布存在明显差异,且以橘芯在440 nm处荧光强度的下四分位数(Q1)与砂囊在440 nm处荧光强度的上四分位数(Q3)的平均值作为分类标准,去囊衣带芯橘瓣检出准确率可达85%。对(370±2)nm激发下得到的单色(440±5)nm荧光图像进行二值化及形态学处理后,可在以砂囊为背景的橘瓣图形中显现橘芯亮斑。利用橘芯与砂囊的荧光特性差异进行机器视觉成像分析,可作为识别去囊衣带芯橘瓣的一种有效方法。     

Origin of the color of Cv. rhapsody in blue rose and some other so-called "blue" roses

Flowers of the rose cultivar Rhapsody in Blue display unusual colors, changing as they age, from a vivid red-purple to a lighter and duller purple, which are based on tonalities corresponding to hue angles between 340 and 320 degrees in the CIELAB scale. Unexpectedly, the chemical basis of these colors is among the simplest, featuring cyanin (cyanidin 3,5-di-O-glucoside), the most frequent anthocyanin in flowers, as the sole pigment and quercetin kaempferol glycosides as copigments at a relatively low copigment/pigment ratio (about 3/1), which usually produces magenta or red shades in roses. This color shift to bluer shades is coupled with the progressive accumulation of cyanin into vacuolar anthocyanic inclusions (AVIs), the occurrence of which increases as the petals grow older. In addition to the normal lambda(max) of cyanin at approximately 545 nm, the transmission spectra of live petals and of epidermal cells exhibit a second lambda(max) in the 620-625 nm range, the relative importance increasing with the presence of AVIs. In petals of fully opened flowers, the only pigmented structures in the vacuoles of epidermal cells are AVIs; their intense and massive absorption in the 520-640 nm area produces a much darker and bluer color than measured for the vacuolar solution present at the very first opening stage. Cyanin is probably "trapped" into AVIs at higher concentrations than would be possible in a vacuolar solution and in quinonoidal form, appearing purple-blue because of additional absorption in the 580-630 nm area. Quite similar pigmentation features were found in very ancient rose cultivars (cv. L'Evêque or Bleu Magenta), also displaying this type of so-called "blue" color.     

Factors influencing phenolic compounds in table olives (Olea europaea)

The Mediterranean diet appears to be associated with a reduced risk of several chronic diseases including cancer and cardiovascular and Alzheimer's diseases. Olive products (mainly olive oil and table olives) are important components of the Mediterranean diet. Olives contain a range of phenolic compounds; these natural antioxidants may contribute to the prevention of these chronic conditions. Consequently, the consumption of table olives and olive oil continues to increase worldwide by health-conscious consumers. There are numerous factors that can affect the phenolics in table olives including the cultivar, degree of ripening, and, importantly, the methods used for curing and processing table olives. The predominant phenolic compound found in fresh olive is the bitter secoiridoid oleuropein. Table olive processing decreases levels of oleuropein with concomitant increases in the hydrolysis products hydroxytyrosol and tyrosol. Many of the health benefits reported for olives are thought to be associated with the levels of hydroxytyrosol. Herein the pre- and post-harvest factors influencing the phenolics in olives, debittering methods, and health benefits of phenolics in table olives are reviewed.     

Effect of cultivar and processing method on the contents of polyphenols in table olives

Polyphenols were determined by HPLC in the juice and oil of packed table olives. The phenolic compositions of the two phases were very different, hydroxytyrosol and tyrosol being the main polyphenols in olive juice and tyrosol acetate, hydroxtyrosol acetate, hydroxytyrosol, tyrosol, and lignans (1-acetoxypinoresinol and pinoresinol) in oil. The type of processing had a marked influence on the concentration of polyphenols in olive juice and little on the content in oil. The analyses carried out on 48 samples showed that turning color olives in brine had the highest concentration in polyphenols ( approximately 1200 mg/kg), whereas oxidized olives had the lowest ( approximately 200 mg/kg). Among olive cultivars, Manzanilla had a higher concentration than Hojiblanca and Gordal. The type of olive presentation also influenced the concentration of polyphenols in olives, decreasing in the order plain > pitted > stuffed. The results obtained in this work indicate that table olives can be considered a good source of phenolic antioxidants, although their concentration depends on olive cultivar and processing method.     

基于叶绿素荧光光谱的生菜硝酸盐含量检测

为了寻求生菜叶片硝酸盐含量的快速无损检测方法,该文利用叶绿素荧光光谱分析技术对生菜(Lactuca sativa L.)叶片硝酸盐含量进行检测研究。对比及分析500~550、650~715和715~800 nm 3个波段的叶绿素荧光光谱特征参数与生菜叶片硝酸盐含量的关系,得出650~715 nm波段的叶绿素荧光光谱特征参数与生菜叶片硝酸盐含量之间线性关系显著,决定系数R2为0.816,标准误差为0.147,以此建立的回归模型能够很好地反映生菜叶片硝酸盐含量与叶绿素荧光光谱特征参数的关系;将同批进行试验的30个样本作为回归方程的校验集,进行模型验证,预测值与实测值之间决定系数R2为0.752,表明回归模型对生菜叶片硝酸盐含量有良好的预测效果。研究结果为生菜叶片硝酸盐含量的快速无损检测提供参考。     

Hydroxytyrosol 4-beta-D-glucoside,an important phenolic compound in olive fruits and derived products

There is increasing interest in olive polyphenols because of their biological properties as well as their contribution to the color, taste, and shelf life of olive products. However, some of these compounds remain unidentified. It has been shown that hydroxytyrosol 4-beta-D-glucoside (4-beta-D-glucosyl-3-hydroxyphenylethanol) coeluted with hydroxytyrosol [(3,4-dihydroxyphenyl)ethanol] under reversed phase conditions in the phenolic chromatograms of olive pulp, vegetation water, and pomace of olive oil processing. A method to separate this compound from hydroxytyrosol by HPLC has been developed. The concentration of this glucoside increased in olive pulp with maturation and could be the main phenolic compound in mature olives. In contrast, the presence of this compound was not detected in olive oil by using HPLC-MS. The compound must be considered both in table olives and olive oil processing because of its glucose and hydroxytyrosol contribution to these products.