Comparative study on phenolic content and antioxidant activity during maturation of the olive cultivar Chemlali from Tunisia

For the first time the identification and quantification of phenolic compounds of the Olea europaea L. cv. Chemlali olive were carried out to examine their profile during maturation. The phenolic composition was studied by using reverse-phase high-performance liquid chromatography during all steps of fruit development. Oleuropein is the abundant phenolic compound in Chemlali olive, and its concentration increases during maturation. An indirect relationship between oleuropein content in olive fruit and hydroxytyrosol was observed. Weak changes in the amounts of the other phenolic monomers and flavonoids were also observed. The total phenolic content varied from 6 to 16 g/kg expressed as pyrogallol equivalents. Its highest level was found at the last maturation period. The antioxidant capacity of olive extracts was evaluated by measuring the radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl. The IC(50) values of the olive extract ranged from 3.2 to 1.5 microg/mL. There was a correlation between antioxidant activity and total phenolic content of samples. The antioxidant activity increased with maturation. This could be attributed to the increase of the tolal phenol level with fruit development.     

Comparative study of virgin olive oil quality from single varieties cultivated in Chile and Spain

Olive tree varieties that were cultivated only in the Mediterranean basin a few decades ago are now planted in the Southern Hemisphere as well. The chemical composition of the oils produced in countries as far distant as Spain and Chile are affected by differences in latitude and climate. In this work, seven monovarietal virgin olive oils from Chile (Arbequina, Barnea, Frantoio, Koroneiki, Leccino, Manzanilla and Picual) have been characterized by the chemical compounds responsible for taste (phenols) and aroma (volatiles). The oils were produced in five regions of Chile, and the concentration values of some chemical compounds were related to the geographical location of the olive tree orchards. Virgin olive oils from the major cultivars, Arbequina and Picual, were characterized in comparison with the same monovarietal oils produced in Spain. The concentration values of fourteen volatile compounds showed significant differences (p < 0.05) between the oils produced in Spain and Chile. Concerning the phenol composition, main differences were found on the secoiridoids derivatives of oleuropein and ligstroside, apigenin and luteolin.     

Comparative study on the triglyceride composition of almond kernel oil. A new basis for cultivar chemometric characterization.

Nine triglycerides (LLL, OLL, PLL, OLO, PLO, PLP, OOO, POO, and SOO; triglycerides are abbreviated using L, O, P, and S for linoleoyl, oleoyl, palmitoyl, and stearoyl fat acid radicals, respectively) in the almond kernel of 19 different cultivars have been determined by high-performance liquid chromatography. Multivariate techniques have been applied to the data from 114 chromatographic determinations. Principal component analysis efficiently reduces the number of variables so that the first two principal components explain 84.4% of the total variance. The classification obtained by the application of cluster analysis to triglyceride composition differentiates the American cultivar Texas from the rest of the cultivars. The Italian cultivars are grouped. The cultivars Achaak, Del Cid, Malague?a, Desmayo Largueta, and Chellaston form another group, and the largest group includes most of the Spanish cultivars. Discriminant analysis provides convenient functions to describe the four groups previously established by cluster analysis. The calculated classification functions correctly assign samples from the testing set to their respective groups.     

Non-concordance between genetic profiles of olive oil and fruit: a cautionary note to the use of DNA markers for provenance testing

To investigate the contribution of paternal alleles to the DNA content of olive oil, genetic analyses of olive DNA samples from fruits, leaves, and oil derived from the same tree (cv. Leccino) were carried out. DNA extracted from maternal tissues--leaves and flesh--from different fruits showed identical genetic profiles using a set of DNA markers. Additional simple sequence repeat (SSR) alleles, not found in the maternal samples, were amplified in the embryos (stone), and they were also detected in DNA extracted from the paste obtained by crushing whole fruits and from the oil pressed from this material. These results demonstrate that the DNA profile obtained from olive oil is likely to represent a composite profile of the maternal alleles juxtaposed with alleles contributed by various pollen donors. Therefore, care needs to be taken in the interpretation of DNA profiles obtained from DNA extracted from oil for resolving provenance and authenticity issues.     

Comparative analysis of soil DNA extraction methods to study various ammonium-oxidizing bacteria and archaea by PCR-DGGE

A comparative analysis of five methods of extraction and purification of soil DNA, including a modification of the authors, was performed for the further molecular investigation of various ammonium-oxidizing bacteria and archaea in soils. Experiments using soil samples from natural ecosystems and agroecosystems of the European area of Russia established that the amount of DNA extracted by different methods depended significantly on the type of soil. The subsequent molecular analysis (PCR-DGGE) of ribosomal (16S rRNA) and functional (amoA) genes demonstrated significant differences in the community structure of ammonium oxidizers depending on the method of DNA extraction. The best results were obtained for acidic soil (soddy-podzolic and gray forest soils) when using the method of Griffiths et al. [4] with our own modification. On the other hand, application of commercial DNA extraction kits was most efficient for soils with a high content of humus (black and chestnut soils). According to the results obtained, molecular analysis of soil microbe communities required selection of optimum conditions for DNA extraction, especially for soils with high contents of organic compounds and clay minerals at different pH levels.     

Phenolic compositions of grapes and wines from cultivar cabernet sauvignon produced in chile and their relationship to commercial value

The phenolic composition of wine depends on, among other factors, the grapes used to make it. In this sense, knowledge of the chemical composition of grapes and its association with the resulting wines is an important tool to determine if there is a relationship between the phenolic composition of grapes and the price that these wines obtain in the market. For this purpose, grape skins and seeds from the cultivar Cabernet Sauvignon from the central region of Chile, in 2009 and 2010 vintages from two ripening points, were subjected to chemical and phenolic analyses, as were the wines made from these grapes. Grapes and the corresponding wines from three retail price wine categories, U.S. $6-8, U.S. $28-30, and U.S. $150-160, were evaluated. No differences were found across the price categories in the chemical analysis of grapes. Berry skins and wines from the higher price categories presented a higher concentration only of total tannins, and the differences in their concentrations were only among the different fractions of proanthocyanidins in the skins, seeds, and wines; there were no differences in their proportions. A seasonal effect influenced the concentrations of certain compounds in grapes and led to a decrease in the concentration of total phenols, total tannins, and total anthocyanins between sampling dates as harvesting moved toward the common commercial grape harvest in Chilean viticulture.     

Novel preparation method of template DNAs from wine for PCR to differentiate grape (Vitis vinifera L.) cultivar

Template DNAs were extracted from wine and purified for use as samples for PCR to differentiate grape cultivars. It has been pointed out that the authentication of grape material by PCR using wine as a material is very difficult. The problems are (1) decomposition of DNAs during fermentation; (2) contamination of DNAs from microorganisms such as yeast; (3) interference of DNA extraction by polysaccharides and polypeptides in the beverages; and (4) coexistence of PCR inhibitors, such as polyphenols. For this study was developed a novel preparation method of template DNA from wine to differentiate grape cultivars using PCR by (1) lyophilizing and pulverizing the fermented beverage to concentrate the DNAs; (2) decomposition of polysaccharides and proteins so as not to inhibit DNA extraction using heat-resistant amylase and proteinase K without DNA damage by endogenous DNase; and (3) separation of the template DNAs for PCR from PCR inhibitors, such as polyphenols, by purification using 70% EtOH extraction and isopropyl alcohol precipitation. To prevent the amplification of microorganisms' DNAs during PCR, suitable PCR primers closely related to the specific plant DNAs, such as chloroplast DNA and mitochondrial DNA, were selected. The sequences of the amplified DNAs by PCR were ascertained to be the same as those of grape materials.     

Comparative study of two methods for extraction of aflatoxin from peanut meal and peanut butter

The difference between the CB and Best Foods methods in extracting aflatoxins from peanut products has been studied. The CB method yields 60, 121, 35, and 22% higher results for aflatoxins B1, B2, G1, and G2, respectively for 4 samples of peanut meal and 6 samples of peanut butter studied. Both reverse phase liquid chromatography and thin layer chromatography were used to quantitate the extracted aflatoxins.     

Solid-phase microextraction in the analysis of virgin olive oil volatile fraction: characterization of virgin olive oils from two distinct geographical areas of northern Italy

SPME was employed to characterize the volatile profile of virgin olive oils produced in two geographical areas of northern Italy: the region of the Gulf of Trieste and the area near Lake Garda. There are as yet no data on the headspace composition of virgin olive oils from these regions, characterized by particular conditions of growth for Olea europaea. Using the SPME technique coupled to GC-MS and GC-FID, the volatile components of 42 industrially produced virgin olive oil samples were identified and the principal compounds quantitatively analyzed. Significant differences in the proportion of volatile constituents from oils of different varieties and geographical origins were detected. The results suggest that besides the genetic factor, environmental conditions influence the volatile formation.     

Comparative study of three different methods for the determination of aflatoxins in tahini

Aflatoxins spiked at three different levels (6.5, 13.0, and 19.5 microg/kg) in tahini, a sesame butter, were analyzed by using three different methods: high-performance liquid chromatography (HPLC), fluorometry, and enzyme-linked immunosorbent assay (ELISA). An immunoaffinity column was used for cleanup and purification of extracts prior to detection by HPLC and fluorometry. All methods were statistically evaluated for accuracy, precision, and simple correlations. Additionally, 14 tahini samples randomly obtained from Turkish retail markets were analyzed using an immunoaffinity column cleanup procedure coupled with the HPLC detection method. The fluorometric determination method involving an immunoaffinity column cleanup step was found to be highly correlated with the HPLC method (r = 0.978). Both methods were found to be effective due to their high recoveries and low variance for the prediction of total aflatoxin contamination in tahini samples. The ELISA method, due to its high variation in replicates, was found to be applicable only as a screening method. The survey study demonstrated the need for control of aflatoxin contamination of foodstuffs involving sesame seeds as an ingredient.     

Application of gas chromatography coupled to triple quadrupole mass spectrometry for the multiresidue analysis of pesticides in olive oil

A new multiresidue method has been developed and validated for the simultaneous determination of 100 pesticide residues in olive oil. The determination of pesticide residues was carried out in only 19 min by gas chromatography coupled to tandem mass spectrometry using a triple quadrupole mass analyzer. The mass spectrometer was operated in electron ionization and the selection reaction monitoring mode was used, acquiring two or three fragmentation reactions per compound. Two extraction processes were studied, and an evaluation of the stability and sensitivity of the chromatographic system has been performed for the tested extraction procedures. The final proposed methodology was based on a liquid-liquid partition with an n-hexane/acetonitrile mixture followed by a gel permeation chromatography cleanup step. An adequate lineal relation was obtained in the studied concentration range (10-200 microg kg (-1)); the recovery values were in the range 70-110% for the two levels of concentration studied: 12 and 50 microg kg (-1). Precision values, expressed as relative standard deviation, were lower than 18% at the aforementioned spiking levels; detection limits, confirmation limits, and quantitation limits were below or equal to 1.9, 2.6, and 3.6 microg kg (-1), respectively. The developed methodology was applied to the analysis of pesticide residues in real samples of olive oil from the south of Spain.     

Comparative study of methods for the extraction of eleven organophosphorus pesticide residues in rice.

Several extraction methods are compared for the simultaneous analysis of organophosphorus pesticides in unpolished rice. Four stationary phases were used for the subsequent gas-liquid chromatographic (GLC) determination of the selected pesticides. Using 3 different GLC columns, 11 pesticides were completely separated and identified. The efficiency of the cleanup and the sensitivity of the analytical method were evaluated by using powdered unpolished rice samples fortified with the pesticides and also wheat and dried bean samples. Average recoveries ranged from 74.7% for disulfoton to 97.4% for malathion in unpolished rice and from 68.1% for disulfoton to 108.3% for malathion in other crops. The method described is applicable to the analysis of selected organophosphorus pesticide residues in unpolished rice, wheat, buckwheat, and dried beans.     

Identification of 9(E),11(E)-18:2 fatty acid methyl ester at trace level in thermal stressed olive oils by GC coupled to acetonitrile CI-MS and CI-MS/MS, a possible marker for adulteration by addition of deodorized olive oil

The olive oil market is suffering from sophisticated illegal treatments. One common adulteration process consists of the addition to virgin olive oil of lower quality oils, such as "lampante" oil, an inexpensive oil and with some organoleptic defects, which is then submitted to thermal deodorization under vacuum processes for removal of the undesired flavor components. Such a blending may not have a huge influence on the chemical composition and may not significantly affect the parameters usually checked as quality indicators, although the organoleptic properties may change. As a consequence, a major effort is being devoted to find reliable markers able to unmask such adulterations. We report here the complete characterization of a compound, detected at trace levels exclusively in thermal stressed oils, which could be a candidate marker for adulteration. The investigation, carried out by GC-MS and GC-MS/MS, provided its complete structure, including the stereochemistry, shown to be a 9(E),11(E)-18:2 fatty acid methyl ester. Experimental data also confirmed the influence of both temperature and heating time on formation and concentration of this compound.     

延河流域三种土壤可蚀性K值估算方法比较

土壤可蚀性因子K是表征土壤侵蚀作用的敏感指标。采用3种土壤可蚀性K值估算方法(Torri.D模型、EPIC模型、Shirazi公式法)对延河流域土壤可蚀性进行对比研究,以实测K值为依据,采用相关性分析和模型估算筛选出符合该区侵蚀特点的土壤可蚀性估算方法。结果表明:延河流域土壤中有机碳、黏粒、粉粒含量随植被覆盖度的变化由北向南逐渐增加,平均质量直径(D_(MW))表现为森林森林草原草原,K_(EPIC)和Kshirazi与D_(MW)呈正相关,而Torri.D模型估算K值(K_(Torri.D))与D_(MW)呈相反的变化趋势,即从草原到森林草原再到森林,土壤团聚体稳定性和抗侵蚀性逐渐增加。K_(Torri.D)的变化范围为0.068~0.147 5,高于真实K值(0.031 2~0.079 6),相比于其他两种方法,Torri.D模型平均绝对误差(MAE)、平均相对误差(MRE)和均方根误差(RMSE)更接近于0,而精度因子(Af)更接近1,具有更高的可信度,更加适用于延河流域土壤侵蚀敏感性评价和土壤流失量预测。     

Real-time TaqMan polymerase chain reaction detection and quantification of cow DNA in pure water buffalo mozzarella cheese: method validation and its application on commercial samples

Mozzarella cheese obtained from buffalo (Bubalus bubalis) milk is a typical Italian product certificated by means of the European Protected Designation of Origin (PDO). Mozzarella cheese can also be obtained from bovine milk or bovine/buffalo milk mixtures, but in this case, it cannot be sold as PDO product, and its label must report the actual ingredients. However, bovine milk in PDO products was frequently detected in the past, suggesting fraudulent addition or accidental contamination. Several methods based on end-point polymerase chain reaction (PCR) have been profitably applied in a large number of tests to detect the presence of undeclared ingredients, also in dairy products. In the present study we report a real-time PCR method able to quantify bovine milk addition to pure buffalo cheese products. We validated a normalized procedure based on two targets: bovine mitochondrial cytochrome b (cyt b) to detect and quantify the bovine DNA and nuclear growth hormone (GH) gene used as a universal reference marker. With the use of this real-time PCR assay, 64 commercial mozzarella di bufala cheese samples purchased at local supermarkets, dairy shops, or directly from cheese manufacturers were analyzed. The results obtained demonstrate that most of the commercial samples were contaminated with bovine milk. Therefore, this assay could be conveniently employed to carry out routine and accurate controls aimed not only to discourage any fraudulent behavior but also to reduce risks for consumer health.     

Comparative study of colorimetric DNA hybridization method and conventional culture procedure for detection of Salmonella in foods

A second generation nucleic acid hybridization assay has been developed and evaluated against the conventional culture method for detection of salmonellae in foods. The assay involves a liquid hybridization with Salmonella-specific oligonucleotide probes, capture of probe:target hybrids onto a solid support (plastic dipstick), and a colorimetric end point detection. The assay can be completed in 2.5 h, following approximately 44 h of culture enrichment. One thousand samples representing 20 food types were analyzed in parallel by both methods. Samples included uninoculated test product, and product inoculated with Salmonella at 2 levels. Eighteen Salmonella serotypes were used as inocula. The data demonstrate that the colorimetric hybridization method and the conventional culture method are equivalent in their ability to detect Salmonella contamination of foods.     

Rapid methods for differentiating reactivated from residual phosphatase in milk and cream: collaborative study.

Three methods for differentiating reactivated from residual phosphatase in milk and cream were collaboratively tested using both magnesium acetate and magnesium chloride for reactivating phosphatase. The methods evaluated were the modified Scharer rapid test, the rapid colorimetric test, and the Rutgers method. Nine collaborators tested 6 unknown milk samples containing reactivated and/or residual phosphatase, and 16 collaborators tested 6 unknown cream samples containing reactivated and/or residual phosphatase. Results indicated that use of magnesium acetate in place of magnesium chloride for reactivating phosphatase improved test results. Visual tests (modified Scharer rapid and Rutgers) predicted correct results when the samples contained high levels of reactivated or residual phosphatase. In borderline cases where the reactivated phosphatase contents of the undiluted control sample and the diluted sample containing Mg were very close, the test results of the visual methods were significantly different from 100% correct results at the alpha=0.05 level. Use of a photoelectric colorimeter or its equivalent for measuring the absorbance in conjunction with the modified Scharer rapid test improved results considerably. The modified Scharer rapid test was adopted official first action.     

Methods of sample preparation for detecting alkaline phosphatase in casein: collaborative study

A collaborative study was conducted in which 2 different sample preparation techniques were used to determine alkaline phosphatase in casein by the rapid colorimetric test. Seven collaborators tested 10 unknown casein products containing different amounts of residual phosphatase. Results indicated that the phosphatase contents of casein prepared by the 2 methods were not significantly different. The collaborators correctly analyzed 100% of the test samples that were ground and 98% of the test samples that were unground. The alternative rapid sample preparation method has been adopted official first action.