Determination of the diglyceride content in greek virgin olive oils and some commercial olive oils by employing (31)P NMR spectroscopy

In this study, the diglyceride contents of 96 samples of virgin olive oils from the regions of Crete, Lesvos, Messinia, Pilion, Zakynthos, Halkidiki, and Ilia, 15 samples of commercial extra virgin and pure olive oils, and 3 samples each of refined olive oils and pomace oils were determined by a facile method introduced in a previous publication. This method is based on the phosphitylation of the free hydroxyls of the diglycerides with 2-chloro-4,4,5,5-tetramethyldioxaphospholane and the integration of the appropriate peaks in the (31)P NMR spectra. This preliminary study showed interesting trends in the diglyceride content of the virgin olive oils from the various regions of Greece that can be used as simple criteria to assess the olive oil characteristics. Analysis of variance has been carried out for the diglyceride content of each region in an attempt to detect possible differences in the diglyceride levels among the various regions. Finally, the relationship between the ratio of 1,2-diglycerides to the total amount of diglycerides and the total amount of diglycerides has been used to monitor the quality of virgin olive oils, commercial olive oils, refined olive oils, and pomace oils.     

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.     

Characterization of fresh and decomposed dissolved organic matter using excitation-emission matrix fluorescence spectroscopy and multiway analysis

Fresh and decomposed dissolved organic matter (DOM) derived from 13 plant biomass and animal manure sources was characterized using multidimensional fluorescence spectroscopy with parallel factor analysis (PARAFAC), high-performance size-exclusion chromatography, and UV-vis spectroscopy. The PARAFAC analysis modeled seven fluorescence components: tryptophan-like, tyrosine-like, and five humic substance-like components. For most of the plant-derived DOM solutions, decomposition significantly affected the concentration of three humic substance-like-associated components, increasing two and decreasing one. The effect of decomposition upon DOM derived from animal manures was dependent on the manure source. For a majority of the DOM extracts, the ratio of fluorescence intensity to absorptivity at 254 nm increased following decomposition, indicating that fluorescing DOM compounds were generally more resistant to biodegradation than nonfluorescing UV-absorbing compounds. Molar absorptivity, humification index (HIX), and apparent molecular weight (MWAP) increased by 38.0, 38.8, and 370%, respectively, following decomposition. Spearman correlation analysis showed a strong positive relationship between the humic substance-like components and the DOM MWAP, absorptivity, and HIX. The results of this study support the use of multidimensional fluorescence spectroscopy with PARAFAC as a method to monitor the decomposition of carbon-rich soil amendments such as crop residues, green manures, and animal manures.     

FTIR spectroscopy and multivariate analysis can distinguish the geographic origin of extra virgin olive oils

This work investigates whether Fourier transform infrared spectroscopy (FTIR), in combination with multivariate analysis, can distinguish extra virgin olive oils from different producing countries. Duplicate spectra were collected from 60 oils from four European countries. Two approaches to data analysis were used as follows: first, the "whole spectrum" method of partial least squares (PLS) followed by distance-based linear discriminant analysis (LDA) applied to the PLS scores, and second, a genetic algorithm (GA) for variate selection from the raw data, followed by LDA applied to the selected subset. The PLS-LDA approach produced a cross-validation success rate of 96%, whereas the GA-LDA approach achieved a 100% cross-validation success rate, from subsets comprising only eight variates. Neither the selected variate nor the whole spectrum approach was able to offer insight into the origin of the discrimination in biochemical terms. However, FTIR analysis is rapid, and this work shows that it has the required discriminatory power to potentially offer a "black box" method of screening oils to verify their country of origin.     

分提橄榄油中主要脂肪酸成分傅里叶变换红外光谱法测量（英）

为了测量从橄榄油中分提的高、低熔点油脂的脂肪酸成分,在4 000～600 cm-1 的范围测量了31个含有不同脂肪酸成分植物油的傅里叶变换红外光谱,用于建立偏最小二乘（PLS）回归分析校正模型。在油脂的傅里叶变换红外光谱变量和脂肪酸组成变量之间建立了交叉验证的PLS校正模型。为了校正油酸和亚油酸含量,在4?000～600 cm-1 的频率范围,经平滑,二阶导数,规范化处理的红外光谱获得了最好的交叉验证校正模型和最佳的预测结果。PLS校正模型预测结果表明,与高熔点橄榄油（油酸,72.29%,亚油酸,9.98%）相比,低熔点橄榄油含有较高的油酸含量和亚油酸含量（油酸,77.46%,亚油酸,12.51%）,预测的结果与气相色谱测量的结果有很好的一致性。建立的PLS校正模型预测橄榄油的不饱和脂肪酸含量具有较好的相关性。该研究为分提油脂质量的判别评价提供了便捷的方法。     

Quality control of commercially available essential oils by means of Raman spectroscopy

The essential oils of different spices and aroma plants were investigated fast and nondestructively by means of Raman spectroscopy. The main ingredients of these oils, mostly monoterpenes and phenylpropane derivatives, were identified. A hierarchical cluster analysis shows that the oils of the investigated plants cluster chemotaxonomically, which means according to their chemical composition and not according to their botanical degree of relationship. In addition, cross-sections of anise seeds were analyzed applying the Raman mapping technique to localize and investigate the distribution of the oil in situ.     

Detection of the presence of refined hazelnut oil in refined olive oil by fluorescence spectroscopy

The fluorescence spectroscopy technique has been tested as regards its ability to differentiate between refined hazelnut and olive oils. Classification of these oils based on their excitation-emission fluorescence spectra data (spectral range 300-500 nm of the excitation spectra at lambdaem=655 and spectral range 650-900 of the emission spectra at lambdaex=50 nm) was performed using principal component analysis and artificial neural networks. Both methods provided good discrimination between the refined hazelnut and olive oils. The results have also pointed out the possibilities of a spectrofluorimetric method joined to multivariate analysis, to differentiate refined oils, and even to detect the presence of refined hazelnut oils in refined olive oils at percentages higher than 9%.     

Rapid quantitative assessment of the adulteration of virgin olive oils with hazelnut oils using Raman spectroscopy and chemometrics

The authentication of extra virgin olive oil and its adulteration with lower-priced oils are serious problems in the olive oil industry. In addition to the obvious effect on producer profits, adulteration can also cause severe health and safety problems. A number of techniques, including chromatographic and spectroscopic methods, have recently been employed to assess the purity of olive oils. In this study Raman spectroscopy together with multivariate and evolutionary computational-based methods have been employed to assess the ability of Raman spectroscopy to discriminate between chemically very closely related oils. Additionally, the levels of hazelnut oils used to adulterate extra virgin olive oil were successfully quantified using partial least squares and genetic programming.     

Dynamics of water extractable organic carbon at a subtropical catchment using fluorescence excitation-emission matrix spectroscopy coupled with parallel factor analysis

The vertical and horizontal movement of water extractable organic carbon (WEOC) has a significant influence on soil organic carbon (SOC) storage and water quality. However, how the amount, components and chemical qualities of WEOC change at horizontal and vertical levels, as well as desorption in the cascade model in subtropical red soils, are not fully understood. We sampled soils from four types of land (Elaeocarpus, Masson Pine, Crape-Myrtle and a deposition site) in a subtropical catchment as replicated cores (n = 96) and investigated changes in the molecular and structural constituents of WEOC using excitation-emission matrix fluorescence and UV–visible absorbance spectroscopy. Our results showed that WEOC concentration changed remarkably in all four plots, and that the concentration coupled with the relative abundance of aromatic compounds decreased with increasing soil depth, whereas low-molecular-weight materials increased with increasing soil depth. Five fluorescent components were identified by parallel factor analysis (i.e., tryptophan-like, tyrosine-like and three humic-substance-like components). The relative abundance of these components suggested that WEOC composition shifted along vertical soil layers. Dynamic variations in fluorescent indices indicated that WEOC from the subsoil has strong biological characteristics. Additionally, there was no significant correlation between WEOC and SOC in subsoil, indicating that much of the WEOC within mineral soils is of local microbial origin rather than from local soil organic matter. Sorption processes alone or stepwise exchange processes of WEOC on the soil minerals do not readily occur, as the red soil in a subtropical area is typically unsaturated and potentially has a large sorption capacity. Once soil carbon is adsorbed, it is difficult to release it into soil water by microbial processing. Data from the deposition site suggest that components of hydrophilic WEOC with low molecular weight are preferentially transported.     

Synchronous fluorescence spectroscopy of edible vegetable oils. Quantification of tocopherols

The study demonstrates the application of front face and right angle synchronous fluorescence spectroscopy for the characterization of edible oils. The method enables monitoring of tocopherols, pheophytins, and other fluorescent components in edible oils. Principal component analysis of synchronous fluorescence spectra revealed sample clustering according to the type of oil. Partial least-squares regression was utilized to develop calibration models between fluorescence spectra and total tocopherol content determined by HPLC. The regression models showed a good ability to predict tocopherol content. The best fitting results were obtained for 1% v/v diluted oils and for bulk samples using the entire spectrum, yielding the regression coefficient, r, of 0.991, and root mean square error of cross-validation of approximately 8%. The results presented confirm the capabilities of the fluorescence techniques as a tool for the analysis of edible oils.     

Excitation-emission fluorescence spectroscopy combined with three-way methods of analysis as a complementary technique for olive oil characterization

This paper shows the potential of excitation-emission fluorescence spectroscopy (EEFS) and three-way methods of analysis [parallel factor analysis (PARAFAC) and multiway partial least-squares (N-PLS) regression] as a complementary technique for olive oil characterization. The fluorescence excitation-emission matrices of a set of Spanish extra virgin, virgin, pure, and olive pomace oils were measured, and the relationship between them and some of the quality parameters of olive oils (peroxide value, K232, and K270) was studied. N-PLS was found to be more suitable than PARAFAC combined with multiple linear regression for correlating fluorescence and quality parameters, yielding better fits and lower prediction errors. The best results were obtained for predicting K270. EEFS allowed detection of extra virgin olive oils highly degraded at early stages (with high peroxide value) and little oxidized pure olive oils (with low K270). The proposed methodology may be used as an aid to analyze doubtful samples.     

Kinetics of diglyceride formation and isomerization in virgin olive oils by employing 31P NMR spectroscopy. Formulation of a quantitative measure to assess olive oil storage history

Diacylglycerol isomers and free acidity were determined for five extra virgin olive oils of different initial acidities by employing a facile (31)P NMR methodology as a function of storage time and storage conditions. The kinetic treatment of the hydrolysis of triacylglycerols (TGs) and the isomerization of 1,2-diacylglycerols (1,2-DGs) to 1,3-diacylglycerols (1,3-DGs) during storage of 18 months at ambient temperature in the dark and light and at 5 degrees C in the dark showed that the isomerization is strongly dependent on the rate of the TGs hydrolysis, the initial free acidity (H(0)) of the virgin olive oil samples, and storage conditions. Although the time-evolution of the diacylglycerols (DGs) depends on the TGs hydrolysis, the ratio D of the concentration of 1,2-DGs to the total amount of DGs was found to be independent of this factor. From the kinetic expression of the ratio D, a quantitative measure was formulated that allows the estimation of the storage time or age of virgin olive oils. Application of this quantitative measure to several olive oil samples of known and unknown storage history resulted in a very good agreement with respect to the actual storage time for up to 10-12 months of storage. For a longer storage period, where the isomerization of DGs is close to its equilibrium state, the calculated age index is only indicative.     

Application of stable isotope ratio analysis to the characterization of the geographical origin of olive oils.

To gain information about the geographical origin of oil samples, measurements of delta(13)C and delta(18)O of the whole oil and some of its fractions have been performed on samples coming from fruits of Olea europaea L. produced in Greece, Morocco, Spain, Italy, Tunisia, and Turkey. The results obtained by applying statistical procedures have given pieces of evidence that oil samples have shown the trend to cluster according to the different climatic areas of growing environment of fruits. Some confusion has been observed for samples coming from neighboring countries having similar climates.     

Nondestructive evaluation of anthocyanins in olive (Olea europaea) fruits by in situ chlorophyll fluorescence spectroscopy

Anthocyanins (Anths) in olive (Olea europaea L.) fruits at different degrees of pigmentation were assessed nondestructively by measuring chlorophyll fluorescence (ChlF). The method is based on the comparison of the ChlF excitation spectra from olives with different pigmentation from green to green-red, reddish-purple, and purple. The logarithm of the ratio between the fluorescence excitation spectra (logFER) from two different colored zones gave the difference in the absorption spectrum between them. The absorbance spectrum derived from the logFER between a red olive and the same olive devoid of the skin showed the typical Anth green band (at 550 nm). It matched that recorded by microspectrophotometry on a single pulp cell and the in vitro absorbance spectrum of the olive skin extract. As expected, the in vivo Anths absorption maximum increased in intensity going from less to more mature olives and was higher in the sun-exposed olive side with respect to the sun-shaded side. Absolute quantitative nondestructive determination of Anths for each olive sample was obtained by the logFER calculated for two excitation wavelengths, 550 and 625 nm, of ChlF at 740 nm. Going from green to purple skin colors, the Log[ChlF(625)/ChlF(550)] was fairly well-correlated to the extract Anths concentration. Finally, the relationship between the Anths and the other main phenolics present in the olives analyzed by high-performance liquid chromatography was evaluated. The main result was a net increase of verbascoside with increasing Anths content. On the basis of our results, the development of a new rapid and noninvasive method for the monitoring of olive development and ripening can be envisaged.     

Determination of geographical origin of olive oils using 13C nuclear magnetic resonance spectroscopy. I - Classification of olive oils of the Puglia region with denomination of protected origin

13C nuclear magnetic resonance spectroscopy was used to classify olive oils from the three production areas of the Puglia region labeled with the "denomination of protected origin" (DPO) Terra di Bari, Colline di Brindisi, and Dauno. High resolution (13)C spectra of 173 olive oil samples were measured, and the intensity data of triacylglycerol resonances were processed by using linear discriminant analysis, which was carried out stepwise for variable selection. The olive oil samples from the DPOs Colline di Brindisi and Terra di Bari were 90% correctly classified, whereas only 74% of "Dauno" DPO oils were classified in the true group. The performance of the discriminant model was verified by applying the cross-validation procedure based on the "leave one out" formalism. The discriminant model was evaluated against a blind test set of olive oils from the three DPO areas. All the oils used for the purpose were correctly assigned to their respective groups, with the exception of the Dauno oil samples based on the Coratina cv. They were misclassified as Terra di Bari oils because of a common monovarietal composition.     

Detecting and quantifying sunflower oil adulteration in extra virgin olive oils from the eastern mediterranean by visible and near-infrared spectroscopy

One hundred and thirty-eight oil samples have been analyzed by visible and near-infrared transflectance spectroscopy. These comprised 46 pure extra virgin olive oils and the same oils adulterated with 1% (w/w) and 5% (w/w) sunflower oil. A number of multivariate mathematical approaches were investigated to detect and quantify the sunflower oil adulterant. These included hierarchical cluster analysis, soft independent modeling of class analogy (SIMCA method), and partial least squares regression (PLS). A number of wavelength ranges and data pretreatments were explored. The accuracy of these mathematical models was compared, and the most successful models were identified. Complete classification accuracy was achieved using 1st derivative spectral data in the 400-2498 nm range. Prediction of adulterant content was possible with a standard error equal to 0.8% using 1st derivative data between 1100 and 2498 nm. Spectral features and chemical literature were studied to isolate the structural basis for these models.     

Classification of edible oils by employing 31P and 1H NMR spectroscopy in combination with multivariate statistical analysis. A proposal for the detection of seed oil adulteration in virgin olive oils

A combination of (1)H NMR and (31)P NMR spectroscopy and multivariate statistical analysis was used to classify 192 samples from 13 types of vegetable oils, namely, hazelnut, sunflower, corn, soybean, sesame, walnut, rapeseed, almond, palm, groundnut, safflower, coconut, and virgin olive oils from various regions of Greece. 1,2-Diglycerides, 1,3-diglycerides, the ratio of 1,2-diglycerides to total diglycerides, acidity, iodine value, and fatty acid composition determined upon analysis of the respective (1)H NMR and (31)P NMR spectra were selected as variables to establish a classification/prediction model by employing discriminant analysis. This model, obtained from the training set of 128 samples, resulted in a significant discrimination among the different classes of oils, whereas 100% of correct validated assignments for 64 samples were obtained. Different artificial mixtures of olive-hazelnut, olive-corn, olive-sunflower, and olive-soybean oils were prepared and analyzed by (1)H NMR and (31)P NMR spectroscopy. Subsequent discriminant analysis of the data allowed detection of adulteration as low as 5% w/w, provided that fresh virgin olive oil samples were used, as reflected by their high 1,2-diglycerides to total diglycerides ratio (D > or = 0.90).     

Geographic classification of spanish and Australian tempranillo red wines by visible and near-infrared spectroscopy combined with multivariate analysis

Visible (vis) and near-infrared (NIR) spectroscopy combined with multivariate analysis was used to classify the geographical origin of commercial Tempranillo wines from Australia and Spain. Wines (n = 63) were scanned in the vis and NIR regions (400-2500 nm) in a monochromator instrument in transmission. Principal component analysis (PCA), discriminant partial least-squares discriminant analysis (PLS-DA) and linear discriminant analysis (LDA) based on PCA scores were used to classify Tempranillo wines according to their geographical origin. Full cross-validation (leave-one-out) was used as validation method when PCA and LDA classification models were developed. PLS-DA models correctly classified 100% and 84.7% of the Australian and Spanish Tempranillo wine samples, respectively. LDA calibration models correctly classified 72% of the Australian wines and 85% of the Spanish wines. These results demonstrate the potential use of vis and NIR spectroscopy, combined with chemometrics as a rapid method to classify Tempranillo wines accordingly to their geographical origin.     

Characterization of commercial kava-kava herbal drug and herbal drug preparations by means of nuclear magnetic resonance spectroscopy

The efficiency of one- and two-dimensional NMR experiments in characterizing the content of the constituents of both herbal drugs and herbal drug preparations is demonstrated for kava-kava. These experiments directly detect active constituents represented by kavalactones in both a finely powdered herbal drug and a commercial extract. In addition, NMR spectroscopy can detect all other compounds present in the extract. As previously evidenced, NMR experiments can represent a generally applicable technique for rapid screening and are a complement to the classical analytical techniques such as high-performance thin-layer chromatography, high-performance liquid chromatography, capillary gas chromatography, and electrophoresis. These experiments can be considered a very simple and fast analytical method to obtain a fingerprint of the herbal drugs and their preparations, and to quantify the content of the active principles of the extract.