Detection of low‐quality extra virgin olive oils by fatty acid alkyl esters evaluation: a preliminary and fast mid‐infrared spectroscopy discrimination by a chemometric approach

A set of eighty‐one extra virgin olive oils (EVOOs) was analysed according to the new quality parameters relative to the total amount of methyl and ethyl esters of fatty acids [Σ (FAMEs + FAEEs)] and the ratio between ethyl and methyl esters [ratio of FAEEs/FAMEs (RFF)]. Acquisition of the mid‐infrared spectra was also performed by Fourier Transform Infrared Spectroscopy (FT‐IR). Chemical and spectroscopy data were chemometrically elaborated, and FT‐IR coupled by Partial Least Square (PLS) methodology was developed. Results were statistically similar to official procedure in terms of analytical performance for Σ (FAMEs + FAEEs) and RFF in EVOOs: a good agreement between predicted and actual values on calibration data sets was found (0.98 and 0.83, respectively) and the limit of quantification was low enough (29.3 mg kg^?1) considering the actual limits for Σ (FAMEs+ FAEEs). This new approach, time‐saving and environmentally friendly, can be considered as a useful tool for screening procedures.     

Low‐concentrated acidic electrolysed water treatment of pork: inactivation of surface microbiota and changes in product quality

The study analysed the effect of low‐concentrated acidic electrolysed water (LCAEW) treatment on meat. Microbiological analysis and colour and sensory quality testing during storage were performed on Longissimus thoracis. FT‐IR and FT‐Raman spectroscopy were used to detect eventual changes in the structure of meat after treatment. Meat samples were sprayed for 120 s with LCAEW (0.001%, 0.01% or 0.1% NaCl solution were electrolysed for 0, 5 or 10 min). The highest reduction in total number of micro‐organisms (3.25 log reduction), yeast and moulds (2.68 log reduction) and psychrotrophs (3.10 log reduction) was observed after spraying the meat samples with 0.1% NaCl electrolysed for 10 min. LCAEW caused a decrease in deoxymyoglobin and metmyoglobin concentration in unstored meat samples. The preliminary sensory studies proved that colour changes are not significant for consumers. The IR and Raman spectra indicate that the structure of compounds of meat tissues are not affected by chlorine and chlorine compounds (LCAEW components). LCAEW has no influence on denaturation of meat protein.     

Application of mid-infrared spectroscopy with multivariate analysis and soft independent modeling of class analogies (SIMCA) for the detection of adulterants in minced beef

Chemometric MID-FTIR methods were developed to detect and quantify the adulteration of mince meat with horse meat, fat beef trimmings, and textured soy protein. Also, a SIMCA (Soft Independent Modeling Class Analogy) method was developed to discriminate between adulterated and unadulterated samples. Pure mince meat and adulterants (horse meat, fat beef trimmings and textured soy protein) were characterized based upon their protein, fat, water and ash content. In order to build the calibration models for each adulterant, mixtures of mince meat and adulterant were prepared in the range 2–90% (w/w). Chemometric analyses were obtained for each adulterant using multivariate analysis. A Partial Least Square (PLS) algorithm was tested to model each system (mince meat + adulterant) and the chemical composition of the mixture. The results showed that the infrared spectra of the samples were sensitive to their chemical composition. Good correlations between absorbance in the MID-FTIR and the percentage of adulteration were obtained in the region 1800–900 cm^− 1. Values of R² greater than 0.99, standard errors of calibration (SEC) in the range to 0.0001–1.278 and standard errors of prediction (SEP estimated) between 0.001 and 1.391 for the adulterant and chemical parameters were obtained. The SIMCA model showed 100% classification of adulterated meat samples from unadulterated ones. Chemometric MID-FTIR models represent an attractive option for meat quality screening without sample pretreatments which can identify the adulterant and quantify the percentage of adulteration and the chemical composition of the sample.     

The physiologic state of Escherichia coli O157:H7 does not affect its detection in two commercial real-time PCR-based tests

Multiplex real-time PCR detection of Escherichia coli O157:H7 is an efficient molecular tool with high sensitivity and specificity for meat safety assurance. The Biocontrol GDS^® and DuPont Qualicon BAX^®-RT rapid detection systems are two commercial tests based on real-time PCR amplification with potential applications for quantification of specific E. coli O157:H7 gene targets in enriched meat samples. However, there are arguments surrounding the use of these tests to predict pre-enrichment concentrations of E. coli O157:H7, as well as arguments pertaining to the influence of non-viable cells causing false positive results. The present study attempts to illustrate the effects of different bacterial physiologic states and the presence of non-viable cells on the ability of these systems to accurately measure contamination levels of E. coli O157:H7 in ground beef. While the PCR threshold cycle (C_T) values of these assays showed a direct correlation with the number of bacteria present in pure cultures, this was not the case for ground beef samples spiked with various levels of injured or healthy cells. Furthermore, comparison of post-enrichment cell densities of bacteria did not correlate with injured or healthy cell numbers inoculated before enrichment process. Ground beef samples spiked with injured or healthy cells at different doses could not be distinguished by C_T values from either assay. In addition, the contribution of nonviable cells in generating positive real-time PCR signals was investigated using both assays on pre-enriched and post-enriched beef samples, but only if inoculated at levels of 10⁶ cells/sample or higher, which are levels not typically seen in ground beef.     

Fourier transform infrared spectroscopy (FTIR) as a tool for discriminating <Emphasis Type="Italic">Salmonella typhimurium</Emphasis> contaminated beef

Detection of beef contamination from harmful pathogens will be helpful in protecting the consumer safety and controlling the outbreaks. In this paper, the potential of Fourier transform infrared spectroscopy (FTIR) was investigated to discriminate the Salmonella contaminated packed beef. A suitable headspace sampling system was designed and used to collect the headspace volatiles from the packed meat to the FTIR gas cell. Spectral signatures of headspace volatiles of meat packages were used to classify the packed meat samples as contaminated or not. FTIR spectrum was divided into several regions in order to reduce the dimensionality as well as to select the regions based on the absorbance properties of various volatiles present in headspace of meat package. Principal component analysis was performed on the entire spectrum (4000–500 cm⁻¹) as well as on the selected sub-regions of entire spectrum. Two statistical classification techniques (linear and quadratic discriminate analysis) were used to develop classification models. The statistical models were validated using bootstrap cross validation technique. The total average classification accuracies were evaluated in terms of coefficient of variance (% CV). Based on the mean of total average classification accuracies and its % CV calculated from five similarly conducted experiments, it was found that the statistical models developed on a part of the spectra (500–850 cm⁻¹) and full spectra (4000–500 cm⁻¹) can be used as potential classification models for non-destructive discrimination of Salmonella contaminated packed beef samples from uncontaminated ones. These results need to be further validated on dataset with larger sample size.     

Wine authentication with Fourier Transform Infrared Spectroscopy: a feasibility study on variety,type of barrel wood and ageing time classification

Wine aging in wooden barrels improves the final products’ sensory profile and increases the price. Among the different types of woods, the International Enological Codex of the International Organisation of Vine and Wine (OIV) approves only the use of Oak and Chestnut, thus producing a need for fast and cost‐effective authentication methods. In this study Fourier Transform (FT)‐mid‐infrared (IR) spectroscopy combined with chemometrics was employed to analyze and discriminate wines aged in barrels made from different type of wood species and in stainless steel tanks, over a period of 12 months. The wines used were made from four indigenous Greek grape varieties, two white and two red. A complete differentiation of the samples was achieved according to grape variety, the container type and the aging time based on two spectral regions (from 1800 to 1500 cm^?1 and from 1300 to 900 cm^?1) of their FT‐IR spectra.     

Determination of 4‐ethylcatechol in a Merlot wine using sensory evaluation and the electronic tongue

The objective of this study was to determine the influence of 4‐ethylcatechol (4‐EC) on the sensory profile of Brettanomyces‐contaminated Merlot wine and evaluate electronic tongue discrimination. Using sensory evaluation panels, the consumer detection threshold (DT) and the consumer rejection threshold (CRT) of 4‐EC were determined in Merlot containing 493, 714, 1035 and 1500 μg L^?1 4‐EC. The DT of 4‐EC in Merlot was 823 μg L^?1, while the CRT was 1323 μg L^?1. The electronic tongue discriminated index (DI = 82%) among the samples, with hierarchical clustering showing a clear distinction between the control sample and the spiked samples. The lowest concentration distinguished by the electronic tongue was 493 μg L^?1, a lower value than the sensory threshold determined. These findings suggest that for the detection of 4‐EC in Merlot, the e‐tongue may be more sensitive than some consumers and the e‐tongue may be a suitable methodology for detection of subthreshold concentrations of chemical compounds in wine.     

Development and evaluation of chitosan and its derivative for the shelf life extension of beef meat under refrigeration storage

The current study was aimed to study the shelf life of beef meat at refrigeration storage using novel chitosan derivative (Ch‐D). Chitosan was modified with antimicrobial monomethyl fumaric acid (MFA). The chemical structure of the resulting material was characterized by FT‐IR and HR‐XRD. The results confirmed the successful synthesis of conjugate sample. For the shelf life study, following lots were used: control (distilled water), chitosan 0.5%, Ch‐D 0.5%. The samples were kept at 4 °C for 16 days and analyzed at 4‐day intervals. Ch‐D treatment significantly reduced the total viable counts, enterobacteriaceae, lactic acid bacteria, psychrotrophic bacteria, and yeasts–moulds when compared with the chitosan and control during refrigeration storage. The peroxide value (PV) for Ch‐D was 0.19 ± 0.04 meq O₂ kg^?1 fat and chitosan was 0.21 ± 0.07 meq O₂ kg^?1 fat and showed no significant difference (P < 0.05) at the end of the storage. Thiobarbituric acid reactive substance (TBARs) value for Ch‐D was 0.48 ± 0.02 mg MDA kg^?1 and chitosan was 0.57 ± 0.09 mg MDA kg^?1, while the carbonyl contents for Ch‐D was 3.16 ± 0.23 nm  mg^?1 protein and chitosan was 3.11 ± 0.16 nm  mg^?1 protein at 16 days of storage. Values for Ch‐D and chitosan treatments were significantly lower (P < 0.05) for all the quality attributes as compared with the control throughout storage. At the end of storage, Ch‐D treatment showed a significant increase (P < 0.05) in lightness (L^* = 41.54 ± 0.09) and yellowness (b* = 2.23 ± 0.04). The redness for control was high (a* = 6.12 ± 0.09) as compared with the treated samples. Based primarily on microbial counts, Ch‐D treatment extended the shelf life of beef meat by about 8 days, maintaining acceptable quality attributes.     

Tolerance evaluation in Salmonella enterica serovar Typhimurium challenged with sublethal amounts of Rosmarinus officinalis L. essential oil or 1,8‐cineole in meat model

The induction of direct bacterial tolerance and cross‐tolerance (NaCl, acid pH, high temperature) in Salmonella Typhimurium ATTC 14028 following the exposure to sublethal amounts of the essential oil from Rosmarinus officinalis L. (ROEO), and its major component 1,8‐cineole (CIN) was evaluated in this study. Direct protection was not induced when cells were exposed to 1/2 MIC and 1/4 MIC of ROEO or CIN in meat broth and in previously irradiated meat ground‐beef. Cells exposed to ROEO or CIN at sublethal amounts did not present cross‐protection to high temperature, lactic acid and NaCl. Likewise, cells progressively subcultured in meat broth containing increasing amounts of ROEO or CIN were able to survive only up to 1/4 MIC for both tested substances. From these results, S. Typhimurium ATCC 14028 was not capable to develop direct or cross‐tolerance when exposed to ROEO or CIN in a meat‐based growth media and was not able to develop direct tolerance in a meat‐based model.     

Effects of meat pH and the initial numbers of spores of Clostridium estertheticum on the development of blown pack spoilage of vacuum‐packaged beef

To understand how the initial numbers of Clostridium estertheticum spores on, and the concentration of glucose in meat affect the development of blown pack spoilage, beef of pH ≤5.6 and of pH ≥5.8 was inoculated with the spores at various numbers, vacuum‐packaged and stored at 2 °C. For beef of pH ≤5.6, the volumes of packs inoculated with ≤10 spores did not change; and packs inoculated with ≥30 spores started swelling after 35 days, and the rate of volume increase increased with increasing number of inoculated spores. For beef of pH ≥5.8, packs inoculated 0, three or ten spores slackened, and packs inoculated with ≥30 spores became swollen at the end of storage, but to a much lesser degree than the corresponding packs of beef of pH ≤5.6. Glucose was reduced by 21 mm and depleted in the rinse fluids from swollen packs of beef of pH ≤5.6 and of pH ≥5.8, respectively.     

Modelling the cross‐contamination of Listeria monocytogenes in pork during bowl chopping

This study has developed a predictive model for the cross‐contamination of pork by Listeria monocytogenes during bowl chopping. The transfer rates of L. monocytogenes were measured in sixteen chopping scenarios based on practical work. Meanwhile, contaminated bowl chopper was cleaned with either a dry rag (DR), warm water (WW) or 70% ethanol + water (EW), respectively. It was showed that significant differences (P < 0.05) were observed among the three cleaning methods on the reduction of L. monocytogenes, the greatest log reduction being achieved by EW. Moreover, the model introduced by a previous study, predicting cross‐contamination of L. monocytogenes during meat slicing, was improved and validated in this study. Verification results showed that the improved model was acceptable for predicting L. monocytogenes cross‐contamination during pork chopping with coefficients of determination (R² > 0.82), accuracy factors (A_f < 1.44), bias factors (B_f < 1.42), and root mean square error (RMSE < 0.99). Furthermore, the modified model might provide an effective tool for assessing the risk of the cross‐contamination of meat products.     

Isolation,characterisation and sulphation of soluble polysaccharides isolated from Cucurbita maxima

Using hot water extraction, a large number of polysaccharides were obtained from Cucurbita maxima. A DEAE‐Sepharose CL‐6B chromatography column was used to isolate the major polysaccharides from C. maxima. Two fractions were obtained (LP2‐1 and LP2‐2). LP2‐1 and LP2‐2 consisted of neutral polysaccharides (MW: 1.02 × 10⁴ and 4.32 × 10⁸ g mol^?1, respectively) comprised mainly of galactose units. Analyses by FT‐IR spectrometry, partial acid hydrolysis, periodate oxidation, Smith degradation and GC‐MS indicated that LP2‐1 consisted of 85.3% (1→4) glycosidic linkages and 1.7% (1→3) or (1→6) glycosidic linkages. The LP2‐1 backbone consisted of (1→4)‐linked galactose units, which occasionally branched at O6 or O3. The branches were composed of (1→4)‐linked galactose and terminated with galactose (13%). Two sulphated derivatives (SLP2‐1 and SLP2‐2) with variable degrees of sulphation (DS) were obtained by the sulphur trioxide–pyridine method, without degradation of the polysaccharide. DS of PL2‐1 and PL2‐2 was 0.19 and 0.20, respectively.     

Nontargeted,Rapid Screening of Extra Virgin Olive Oil Products for Authenticity Using Near‐Infrared Spectroscopy in Combination with Conformity Index and Multivariate Statistical Analyses

A rapid tool for evaluating authenticity was developed and applied to the screening of extra virgin olive oil (EVOO) retail products by using Fourier‐transform near infrared (FT‐NIR) spectroscopy in combination with univariate and multivariate data analysis methods. Using disposable glass tubes, spectra for 62 reference EVOO, 10 edible oil adulterants, 20 blends consisting of EVOO spiked with adulterants, 88 retail EVOO products and other test samples were rapidly measured in the transmission mode without any sample preparation. The univariate conformity index (CI) and the multivariate supervised soft independent modeling of class analogy (SIMCA) classification tool were used to analyze the various olive oil products which were tested for authenticity against a library of reference EVOO. Better discrimination between the authentic EVOO and some commercial EVOO products was observed with SIMCA than with CI analysis. Approximately 61% of all EVOO commercial products were flagged by SIMCA analysis, suggesting that further analysis be performed to identify quality issues and/or potential adulterants. Due to its simplicity and speed, FT‐NIR spectroscopy in combination with multivariate data analysis can be used as a complementary tool to conventional official methods of analysis to rapidly flag EVOO products that may not belong to the class of authentic EVOO.     

Microbiological and physicochemical properties of dry‐cured neck inoculated with probiotic of Bifidobacterium animalis ssp. lactis BB‐12

The effect of inoculum level of Bifidobacterium animalis ssp. lactis BB‐12 probiotic strain and ripening period on the quality of dry‐cured neck was studied. The microbiological parameters (Enterobacteriaceae, LAB and TVC) and physicochemical attributes (pH value, a_w, TBARS index, colour) were determined directly after fermentation at 15 °C for 3 weeks, after 6 and 12 months of ripening at 4 °C. The highest LAB count and a lower pH value were found in the meat inoculated with probiotic strain at 6.6 log cfu g^?1 (B2) followed by inoculation with probiotic strain at 6.3 log cfu g^?1 (B1). Level of inoculation had not had an influence on water activity, TBARS index and total colour parameters. Changes of fat oxidation during half‐year of ripening were limited in probiotic meat samples compared to naturally fermented control meat (C). Based on the results, it can be concluded that the most favourable physicochemical and microbiological parameters of the dry‐cured neck were obtained after 6 months of ripening. At that time, the Bifidobacterium BB‐12 at both levels is a good potential starter for meat fermentation.     

Detection of Salmonella spp., Yersinia enterocolitica,Listeria monocytogenes and Campylobacter spp. by real‐time multiplex PCR using amplicon DNA melting analysis and probe‐based assay

Syto9 and probe‐based multiplex real‐time PCR assays for simultaneous detection of a group of foodborne pathogens (named SYLC group), targeting Salmonella spp. (invA gene), Yersinia enterocolitica (ystA gene), Listeria monocytogenes (hly gene) and Campylobacter spp. (rrna gene), have been developed. The Syto9 assay generates amplicon DNA melting curve with four peaks of 86.5 ± 0.5, 84 ± 0.5, 81.5 ± 0.5 and 90.5 ± 0.5 °C corresponding Salmonella spp., Y. enterocolitica, L. monocytogenes and Campylobacter spp. targets, respectively. The sensitivities of the Syto9 and TaqMan assays in artificially inoculated chicken wing rinses were in a range of 3.2 × 10² to 3.1 × 10⁴ and 9.8 × 10² to 1.9 × 10⁴ colony‐forming units per millilitre, respectively, depending on the pathogen. All tested target strains (n = 100) were correctly detected by the both assays, whereas nontarget strains (n = 100) demonstrated no cross‐reactivity representing 100% specificity. The assays are suitable for application in qualitative and quantitative detection of SYLC group pathogens in food matrices.     

Anti‐inflammatory properties of high pressure‐assisted extracts of Grifola frondosa in lipopolysaccharide‐activated RAW 264.7 macrophages

The aim of this study was to determine the in vitro anti‐inflammatory properties of the shake extract (SE) and the high pressure‐assisted extract (PE) of the mycelia of Grifola frondosa in a lipopolysaccharide‐stimulated RAW 264.7 macrophage model. The content of total polysaccharides and β‐glucans of PE at 600 MPa (PE‐600) was 41.2 and 6.2 mg g^?1 dry weight, respectively, which were significantly higher than SE extracts. The results showed that treatment with 500 μg mL^?1 of PE by 600 MPa (PE‐600) did not reduce RAW 264.7 cell viability but did significantly inhibit the production of LPS‐induced NO, PGE₂ and intracellular ROS. The PE‐600 inhibited the activation of NF‐kB and then reduced the production of LPS‐induced TNF‐α, IL‐6 and IL‐1β in a dose‐dependent manner. Thus, the PE could be used as an alternative extraction method for improving the extraction efficacy of G. frondosa and serve as an alternative source of anti‐inflammatory agents.     

A simple sample preparation for simultaneous determination of chloramphenicol and its succinate esters in food products using high-performance liquid chromatography/high-resolution mass spectrometry

ABSTRACT

A simple method is described for the determination of chloramphenicol and its succinate esters in food products. Examination of food products using high-performance liquid chromatography/high-resolution mass spectrometry showed the presence not only of chloramphenicol but also of its succinate forms. A scheme is proposed for determining chloramphenicol and its succinate esters (calculated as chloramphenicol) in meat (beef, pork, poultry), milk, liver, kidney, eggs, fish and honey. Analytes are extracted from a 1.0 g sample with 5 ml acetonitrile. It was found that using the method of standard addition and diluting the extract with water leads to the elimination of matrix effects and also eliminates errors associated with peak splitting due to the separate elution of the differing forms of the analyte. Validation results were satisfactory, with recoveries from 85% to 111% (meat, milk, liver, kidney, eggs, fish and honey) and a relative standard deviation (RSD) lower than 13% for spiked levels of 0.3, 1.0 and 5 µg kg^–1. The limits of detection and quantification (calculated as chloramphenicol for all forms) were 0.1 and 0.3 µg kg^–1, respectively. The RSD of the results of the analysis was < 10%. The duration of the analysis was less than 1 h.     

Detection of aflatoxins in tea samples based on a class‐specific monoclonal antibody

An enzyme‐linked immunosorbent assay (ELISA) using a monoclonal antibody was established to detect aflatoxin B₁ (AFB₁) in tea. The antibody was prepared from a hybridoma derived by fusing Sp2/0‐Ag14 myeloma cells and immunised spleen B cells. The effects from pH, ionic strength, and organic solvents on immunoassay were optimised and the 50% inhibition (IC₅₀) value was 0.057 ± 0.007 ng mL^?1. Spiked black and green tea samples at 10, 20 and 50 ng g^?1 levels of AFB₁ were detected with this proposed ELISA. The recoveries for black tea samples ranged from 68.5% to 117.7% and 73.5 to 114.3% for green tea samples. This immunoassay showed no cross‐reactions with other mycotoxin family but good recognition with related aflatoxins. These results indicate that the ELISA assay could be used as a screening method for aflatoxin detection in tea samples.     

Thermo‐rheological and functional properties of gamma‐irradiated wholewheat flour

The effects of different doses (0, 0.5, 1, 2.5, 5 and 10 kGy) of gamma irradiation on the thermal, rheological and functional properties of the wholewheat flour were evaluated. Water and oil absorption capacity of the flour increased from 85.92% to 91.44% and 1.10 to 1.91 g g^?1 of flour, respectively, with increase in irradiation dose. The dough development time decreased with dose from 4.0 to 3.0 min. The transition temperatures (T_o, T_p and T_c) decreased as the dose increased; enthalpy of gelatinisation (?H) decreased from 5.18 to 4.27 J g^?1 with dose. The flow behaviour showed a shear‐thinning behaviour, and the hysteresis area decreased with dose. The structural recovery decreased with dose. FTIR did not show formation of any new chemical groups.