Phytosterol Determination and Method Validation for Selected Nuts and Seeds

A method involving alkali and/or acid hydrolysis of phytosterols followed by trimethylsilyl ether derivatization coupled with GC-FID analysis was validated and applied in the analysis of major phytosterols (campesterol, stigmasterol, β-sitosterol, and Δ⁵-avenasterol) in nuts (n = 7), seeds (n = 9), legumes (n = 2), and grain (n = 1). The acid-labile Δ⁵-avenasterol was extracted with alkaline hydrolysis only before derivatization. Quantification of all phytosterols was done using the computed relative response factor of 5α-cholestane (internal standard). Analyses of internal and external phytosterol standards showed good linearity for all phytosterols (R ² of 0.999); LOD and LOQ of phytosterols were determined to be 0.01–0.12 and 0.04–0.40 mg/100 g, respectively. Repeatability and reproducibility precision analyses showed acceptable coefficient of variation of less than 3 and 4%, respectively, and satisfactory Horwitz ratio values of <1.0. Excellent accuracy was proved by the high recovery values of 91.4–106.0% for campesterol, β-sitosterol, and stigmasterol. Δ⁵-Avenasterol, the most oxidation-susceptible sterol, showed a recovery of about 60%. The total phytosterol (sum of major phytosterols quantified) contents in the 19 samples varied from 38.8 mg/100 g (white quinoa seed) to 246.2 mg/100 g (sunflower seed). β-Sitosterol was the predominant phytosterol (54–86.0% of total) among all samples except fennel seed in which stigmasterol was predominant. Analytical quality control chart maintained during the study period showed that assays were performed under control. Method validation indicated that the analytical method can be applied for accurate determination of campesterol, β-sitosterol, and stigmasterol in selected food samples.     

High-Performance Liquid Chromatography with Fluorescence Detection for Simultaneous Analysis of Phytosterols (Stigmasterol, β-Sitosterol,Campesterol, Ergosterol,and Fucosterol) and Cholesterol in Plant Foods

A method based on high-performance liquid chromatography (HPLC) with fluorescence (FL) detection for the simultaneous analysis of phytosterols (stigmasterol, β-sitosterol, campesterol, ergosterol, and fucosterol) and cholesterol was developed. To fluoresceinate the sterols, they were derivatized by 1-anthroyl cyanide to the hydroxyl group at carbon 3 of each sterol skeleton. This HPLC-FL method consists of a C-30 column, an isocratic solution using acetone/acetonitril/hexane/water (71:20:4:5, v/v) as the mobile phase at 1.0 mL min^?1 and fluorescence detection at an excitation of 370 nm and an emission of 470 nm. The separation of five phytosterols, cholesterol, and 1-hexacosanol as an internal standard was achieved with sufficient reproducibility and quantitative ability. Our method could evaluate the sterols of land plants such as wood ear fungus, soybean, and parsley, as well as marine algae such as Hiziki (Phaeophyta), Ogonori (Rhodophyta), and Heraiwazuta (Chlorophyta). As a result of the analysis of land plants, wood ear contained a large amount of ergosterol as a precursor of vitamin D₂. Soybean contained a large amount of stigmasterol, campesterol, and β-sitosterol. Parsley contained small amounts of these sterols compared with wood ear and soybean. Among the marine algae, Hiziki, Ogonori, and Heraiwazuta contained large amounts of fucosterol, cholesterol, and β-sitosterol, respectively. The compositions of marine algae differed from those of land plants.     

Development of a Method for the Analysis of Sterols in Sterol-Enriched Deli-Style Turkey with GC-FID

Plant sterols have been recognised by the European Food Safety Authority for their cholesterol-lowering properties and are currently added to several food formulations. The objective of this study was to develop a method based on formation of sterol trimethylsilyl ether derivatives and separation and quantification by GC for the determination of three phytosterols (β-sitosterol, stigmasterol and campesterol) in sterol-enriched deli-style turkey. The assay was linear (concentration range 4.3–172.1 μg/ml, R ²?≥?0.9868) and accuracy and precision were within the acceptance criteria of the U.S. Food and Drug Administration (USFDA) guidelines for method validation, set at <20 % RSD at the lower limit of quantification (LLOQ) and <15 % RSD for all other standards. Accuracy measured by relative response factor (RRF) also met the USFDA validation criteria. No matrix effects were observed. The response factors (RFs) of the three sterols differed significantly to that of the internal standard (ISTD) used, leading to RRF dissimilar to 1 (campesterol?=?1.0167, stigmasterol?=?1.4458, β-sitosterol?=?0.9029). This method is suitable for quantification in meat matrix, and it has been successfully applied to the determination of sterols in sterol-enriched deli-style turkey (21 mg sterols/0.5 g sample).     

Enterohemorrhagic <Emphasis Type="Italic">Escherichia coli</Emphasis> (EHEC) in water from karst springs: detection with real-time PCR and isolation of strains

Within 2 months, two water sources in a karst area in Switzerland were sampled 9 times each, and analyzed by real-time PCR for 6 EHEC O-types, Shiga-like-toxin (stx1 and stx2) and intimin (eae) genes. With the exception of O111, 5 O-types were recorded regularly and at high frequencies (O26: 33.3 %; O157: 33.3 %; O104: 66.6 %; O103: 72.2 %; O145: 94.4 %). Genes for Shiga-like-toxins and intimin were almost omnipresent (stx1: 77.8 %; stx2: 83.3 %; eae: 77.8 %). Strain isolation was undertaken for O-groups 26, 103, 104, 145 and 157. Sample selection for strain isolation was based on Cq-values for the O-groups and stx1, stx2 and eae. From selected samples, frozen enrichment cultures were cultivated on EHLY-agar and 50 typical colonies screened for the O-type and genes encoding for stx1, stx2 and eae. With this approach, only one virulent EHEC-strain could be isolated (Escherichia coli O103, stx1 +; stx2 ?; eae +). We carried out one extensive testing with 800 colonies of O-group O145, and no virulent strain was isolated. Our findings showed that PCR-results are not sufficient to formulate epidemiological conclusions and that the isolation of strains is necessary. However, as the detection procedure of EHEC in foods is cumbersome and expensive, the appropriateness of such an approach in official food control is a matter of debate.     

Development and characterization of reconstituted hydrogel from <Emphasis Type="Italic">Aloe vera</Emphasis> (<Emphasis Type="Italic">Aloe barbadensis</Emphasis> Miller) powder

Structural and rheological characterization of reconstituted hydrogels developed from A. vera non-fibrous alcohol insoluble residue (NFAIR) powder using different methods [viz., shaking (S), heating-shaking (HS), and heating (H)] and concentrations (viz., 0.2–1.6 %, w/v) was carried out. Functional group distribution by FTIR spectroscopy and Congo red (CR) method revealed the presence of acetylated acemannan in A. vera powder. Dynamic oscillation studies of A. vera (NFAIR) fluids at all concentrations of 0.2–1.6 %, w/v, showed gel strength in the order of H > HS > S method. However, in H method, increase in concentration from 0.2 to 1.6 %, w/v showed the conformational transition from semi-diluted solution to weak gel nature. Rheological models described the effect of heating temperatures (HT); 30–90 °C, and times (Ht); 15–60 min on viscoelastic behavior in reconstituted A. vera fluids. The reconstituted A. vera hydrogel prepared with a concentration of 1.6 %, w/v using 50 °C (HT) and 30 min (Ht) condition showed a good agreement with the Power law (storage modulus, G′) and Weak gel model (complex modulus, G*) fitted data (R² > 0.94) resulting higher viscoelastic moduli intercepts; G′₀ (71.5 Pa s ^n′), G″₀ (33.5 Pa s ^n″), lower slopes; n′ (0.22), n″ (0.06), higher network strength (A _F , 121.3 Pa s^1/z ) and number of network (z, 5.3) values. The obtained results suggested that heating at 50 °C/30 min can develop aqueous weak gel networks of A. vera with enhanced gel strength which may be utilized as a novel gelling agent for wide variety of targeted applications in food and pharmaceutical sectors.     

Smart NMR Method of Measurement of Moisture Content of Vegetables During Microwave Vacuum Drying

Microwave drying is usually combined with vacuum environment in conjunction with hot air flow to draw the moisture rapidly. The moisture content of the vegetables undergoing drying is hard to measure online. This research designed a microwave vacuum drying (MVD)-low-field nuclear magnetic resonance (NMR) smart device and investigated the feasibility of NMR method for online measurement of state of moisture during MVD. The relation between the signal amplitude (A ₂) and the true moisture content (M ₁) of six kinds of vegetables (mushroom, carrot, potato, lotus, edamame, vegetable corn) was fitted to estimate if NMR can measure the M ₁ of vegetables directly. Results showed that A ₂ and M ₁ of different fresh vegetables had no single empirical mathematical model to fit. However, for each kind of these vegetables, the A ₂ and corresponding M ₁ in different MVD stages showed a significant linear relationship. The predicted moisture content (M ₂) of mushroom: M ₂ = 5.25351 × 10^?4 A ₂ ? 0.34042, R = 0.996; carrot: M ₂ = 5.78756 × 10^?4 A ₂ ? 0.14108, R = 0.998; potato: M ₂ = 3.10019 × 10^?4 A ₂ ? 0.10612, R = 0.991; lotus: M ₂ = 2.32415 × 10^?4 A ₂ ? 0.01573, R = 0.998; edamame: M ₂ = 3.13310 × 10^?4 A ₂ ? 0.4198, R = 0.996; vegetable corn: M ₂ = 1.69461 × 10^?4 A ₂ ? 0.09063, R = 0.995. The linear models between M ₂ and A ₂ were able to estimate the end point (M ₁ < 8%) of MVD with a high accuracy (P > 0.950).     

Development of a Simple,Rapid Multiplex PCR Method Using the 16S rRNA Gene to Determine the Country of Origin of Brackish Water Bivalve <Emphasis Type="Italic">Corbicula japonica</Emphasis>

In this study, a multiplex PCR detection method was developed to identify the country of origin of Corbicula japonica (clams), a commercially important bivalve in Asia. Specific primer sets that have a single nucleotide mismatch at the 3′ terminus were designed after sequencing the mitochondrial 16S rRNA gene of clams identified as C. japonica originating from Korea, China, and Japan. Using this method, each origin was clearly identified based on the PCR products: three bands for Korean C. japonica (100, 283, and 384 bp), one band for Chinese C. japonica (384 bp), and two bands for Japanese C. japonica (384 and 100 bp). These results indicate that the 16S rRNA gene, which is usually used to identify species, can distinguish the country of origin within C. japonica. Our multiplex PCR assay should be a useful tool for the fair trade of the species.     

Detection of Viable <Emphasis Type="Italic">Vibrio cholerae</Emphasis> Cells in Seafood Using a Real-Time Visual Loop-Mediated Isothermal Amplification Combined with Propidium Monoazide

Vibrio cholerae is an important foodborne pathogen causing severe intestinal infectious diseases that have high incidence and mortality. Almost all of rapid testing methods including immunological and molecular assays for V. cholerae are incapable of distinguishing live cells from dead ones, which may overestimate the number of bacteria and result in many false positive results. To address the problems, live cell-specific dye such as propidium monoazide (PMA) is employed. The loop-mediated isothermal amplification (LAMP) assay is a nucleic acid amplification method that is fast, specific, and sensitive. In this study, we developed a real-time visual LAMP assay using PMA dye to detect thyA gene, thereby identifying viable V. cholerae cells. The results showed that only V. cholarae strains could be detected, and there was no cross-reaction with non-V. cholarae strains. Besides, the sensitivity of the PMA-LAMP assay was 1.1 × 10² CFU/mL and the entire reaction could be accomplished within 1 h. The sensitivity was on par with that of the PMA-qPCR assay. The detection limit in different artificially inoculated samples was 5 CFU/25 g materials for the tested pathogens. In the practical test, the PMA-LAMP assay performed well in comparison with PMA-qPCR and the culture method. Hence, PMA-LAMP assay can provide a highly effective and rapid approach for detecting viable V. cholerae.     

Effects of organic acid pretreatment on microstructure,functional and thermal properties of unripe banana flour

Starch availability has been implicated in unripe matured banana (Musa species), which when processed yields flour suitable for application in low gluten and composite wheat formulations. Unripe Musa species: Williams, Luvhele, Mabonde and Muomva-red obtained from fruit bunch were pretreated with ascorbic, citric and lactic acids, processed into 50 g of flour and characterised for their functional and thermal properties. Scanning electron microscope of unripe banana flour (UBF) showed varying micrographs of flour, with polygonal for Luvhele, oval for Mabonde, elongated for Muomva-red and between polygonal and spherical for Williams. The bulk density of UBF samples was within the range of 0.66–0.84 g/mL for all organic acid pretreatment while citric acid pretreated UBF had the least browning index. Significant difference (p < 0.05) was recorded in swelling power with no significant difference in water solubility index except for Mabonde UBF. Thermal properties showed single endothermic transition for all UBF samples at various pretreatment concentration. The onset temperature (To) of UBF ranges from 49.82 to 65.59 °C, peak temperature (Tp) from 60.11 to 76.71 °C, conclusion temperature (Tc) from 70.36 to 94.16 °C and enthalpy of gelatinization (ΔH) from 2.61 to 32.24 J/g. Short amylopectin chains present in starch of UBF was attributed to low To, Tp, Tc and ΔH values recorded for Mabonde cultivar, while the contribution of heat-moisture treatment rather than organic acid pretreatment of UBF samples was attributed to different gelatinization and transition temperatures recorded for all cultivars examined.     

Determination of Seven <Emphasis Type="Italic">N</Emphasis>-nitrosamines in Agricultural Food Matrices Using GC-PCI-MS/MS

The quantitative analytical methods for seven N-nitrosamines including N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosodiethylamine (NDEA), N-nitrosodibutylamine (NDBA), N-nitrosopiperidine (NPIP), N-nitrosopyrrolidine (NPYR), and N-nitrosomorpholine (NMOR) were established for agricultural food matrices. Four food matrices were used for the method development: rice soup as a fatless solid matrix, apple juice as a fatless liquid matrix, corn oil as a fat-rich liquid matrix, and 20 % alcohol as an alcohol matrix. A combination of solid-supported liquid-liquid extraction (SLLE) using Extrelut NT and a solid phase extraction (SPE) using Florisil was employed for fatless matrices. For an alcohol matrix, only SLLE was used without SPE, and liquid-liquid extraction (LLE) was established for a fat-rich matrix. The extract was analyzed by gas chromatography-positive chemical ionization-tandem mass spectrometry (GC-PCI-MS/MS) using ammonia gas as an ion source. Linearity, recovery, repeatability, inter-day precision, reproducibility, and uncertainty were evaluated for method validation using four matrices. Method detection limits for all of the investigated N-nitrosamines were ranged from 0.10 to 0.18 μg/kg for the rice soup, from 0.10 to 0.19 μg/kg for the apple juice, 0.10 μg/kg for the corn oil, and from 0.10 to 0.25 μg/kg for 20 % alcohol, depending on N-nitrosamines. Established methods were applied to determine seven N-nitrosamines in some agricultural food products.     

Uncertainty of Trypsin Inhibitor Activity Measurement of Legume Crops Using Microtiter Plate Method

Trypsin inhibitors could limit utilization of legumes in human nutrition, but they could also have beneficial health effects. The objective of this study was to measure trypsin inhibitor activity (TIA) of different legumes using microtiter plate method and to identify factors that contribute to uncertainty of TIA measurement. TIA measurements were performed on seeds of faba bean, pea, common vetch, soybean, and common bean cultivars. The significant effect of legume crop on TIA measurement uncertainty was confirmed with P = 0.045. Certain sources of measurement uncertainty were related with the content of trypsin inhibitors (Tis) in legume seeds. In respect to that, significant effect of level of sample dilution (P ? 0.001) was confirmed. Significant influence of the repeated absorbance measurement of sample reaction mixture on uncertainty of TIA measurement was identified (P ? 0.001), and it took 60% of overall TIA measurement uncertainty for soybean cultivars. TIA of soybean cultivars exceeded 90 TUI/mg. Repeated absorbance measurement of positive control reaction mixture took 70% of TIA measurement uncertainty of cultivars with TIA lesser than 4.5 TUI/mg. Graduated cylinder used for preparation of the final sample solutions took the range from 45 to 90% of overall TIA measurement uncertainty of the cultivars whose TIA were in the middle of previously mentioned. The uncertainty of TIA measurement of legume crops was not studied before; thus, this study pointed out that acquiring insight into factors contributing to uncertainty of TIA measurement could give directions for improvement of TIA testing if microtiter plate method is used.     

Quantification of <Emphasis Type="Italic">Penicillium nalgiovense</Emphasis> on Dry-Cured Sausage ‘Salchichón’ Using a SYBR Green-Based Real-Time PCR

To evaluate the effective implantation of a specific protective culture of Penicillium nalgiovense, a real-time quantitative PCR (qPCR) using SYBR Green methodology was developed. Two specific primers were designed on the basis of the published partial sequences of the Internal Transcribe Spacer (ITS)1–5.8S-ITS2 region of various strains of P. nalgiovense. Using the developed method, a PCR product of 51 bp with a T _m value 81.34 °C was detected. T _m values of the amplified product allowed specific differentiation between P. nalgiovense and the remaining mould species tested. The developed qPCR method was tested on inoculated slices of dry-cured sausage (‘salchichón’) showing an efficiency of 97.24 %, a R ² value of 0.99 and a detection limit of P. nalgiovense of 1 log colony-forming units (cfu)/cm². The qPCR method demonstrated that the protective strain of P. nalgiovense grew and competed against an ochratoxin A (OTA)-producing Penicillium verrucosum strain on commercial dry-cured sausage. This qPCR method provides a specific, accurate and sensitive detection and quantification of P. nalgiovense on dry-cured sausage salchichón in order to estimate its colonization during their processing. This assay will improve strategies to prevent and control unwanted mould colonization and OTA risk in dry-cured meat commodities.     

Steaming time effects on the moisture migration and structural properties of Chinese Northern-style steamed bread

The aim of this study was to investigate the steaming time effects on proton transverse relaxation behavior with low field 1H nuclear magnetic resonance and structural properties of Chinese Northern-style steamed bread (CNSB). Three proton populations could be distinguished at the first 4 min: T_2b (0.1–1 ms) corresponded to rigid and exchangeable protons; T₂₂ (9–21 ms) was associated with the water protons in small and large meshes of the dough microstructure; T₂₃ (69–300 ms) was assigned to the water protons on the surface of samples. The starch gelatinization began and the water turned into the integral part of the biopolymer at 6 min, forming T₂₁ (1–3 ms) fraction. The gelatinization effect was strengthened up to 8 min and supplied a more mobile microenvironment, resulting in the increase of T₂₁, A₂₁ and M₂₁. However, the gelatinization process ended at 8 min, bringing about the stabilization of T₂₁, A₂₁ and M₂₁ until 25 min. T₂₂ fraction accounted for the largest proportion during all the steaming process. All variation trends on structural properties of CNSB and T₂ relaxation parameters including T_i, A_i (relative intensity of T_i), and M_i (population abundance of T_i) indicated that 6 and 8 min were the two transitions. The gluten matrix began to be disrupted at 6 min and was quite damaged up to 8 min by scanning electron microscopy. The peaks at 15°, 18°, 20°, and 23° in X-ray diffraction patterns appeared in the first 6 min but were lost up to 8, 10, and 25 min.     

Comparative Study on the Inactivation and Photoreactivation Response of <Emphasis Type="Italic">Listeria monocytogenes</Emphasis> Seafood Isolates and a <Emphasis Type="Italic">Listeria innocua</Emphasis> Surrogate after Pulsed Light Treatment

The inactivation and photoreactivation response of six seafood-isolated Listeria monocytogenes and one Listeria innocua strain after pulsed light (PL) treatment was evaluated. The lower inactivation levels found after exposure of treated samples to daylight during the first 90 min of storage confirmed that both L. innocua and L. monocytogenes have the capability to photorepair PL-induced DNA damage upon appropriate conditions. Photoreactivation levels from 0.2 to 2.1 log CFU cm^?2 were observed depending on treatment intensity (fluence) and Listeria strain. Complete photorepair of PL-caused damage was not found even after treatments inducing low inactivation levels. Photoreactivation increased up to 2.1 log with the applied fluence up to a threshold able to cause between 2.4 and 5.4 log reductions under dark storage. Photorepair was not avoided but lower photoreactivation was observed after higher fluence inducing more than 6 log reductions under dark storage. Both L. innocua and L. monocytogenes serotype 1/2b exhibited the highest photoreactivation levels whereas serotypes 1/2a showed the lowest ones. The overall inactivation and photoreactivation responses of tested Listeria strains were comparable indicating that L. innocua may be a good surrogate for the safe evaluation, optimization and validation of PL technology to control L. monocytogenes in food products and food processing facilities.     

A Rapid and Visual Single Primer Isothermal Amplification-Based Method for the Detection of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> in Raw Pork Products

Staphylococcus aureus (S. aureus) is an important food-borne pathogen which poses a severe threat to public health worldwide. Rapid detection of S. aureus with high sensitivity is of particular importance for food safety. In this study, a novel single primer isothermal amplification (SPIA) method was established to detect S. aureus in food, targeting the accessory gene regulator (agr) gene with a DNA/RNA primer. The developed SPIA method has the advantages of visualization and avoiding tedious electrophoresis. In order to confirm the specificity of this method, 7 S. aureus strains and 26 non-S. aureus strains were detected with their pure cultures. The sensitivity and detection limit of S. aureus with artificially inoculated raw pork products by SPIA were evaluated through fluorescence and turbidity by naked eye and the amplification curve, which were 4.3?×?10⁰ CFU/mL and 5.6?×?10⁰ CFU/g, respectively. Compared with the conventional PCR method, the SPIA has 100-fold higher sensitivity and 100-fold lower detection limit. Therefore, the developed SPIA method is a potentially reliable tool for rapid and visual detection of S. aureus in food.     

Effect of Thermosonic Pretreatment and Microwave Vacuum Drying on the Water State and Glass Transition Temperature in <Emphasis Type="Italic">Agaricus bisporus</Emphasis> Slices

This study aimed to understand the micromechanism of thermosonic pretreatment and microwave vacuum drying on Agaricus bisporus. The water state and glass transition temperature (T _g ) of fresh and thermosonically treated Agaricus bisporus slices during microwave vacuum drying were studied using differential scanning calorimetry (DSC), low-field nuclear magnetic resonance (LF-NMR), and magnetic resonance imaging (MRI). Results showed that four population groups were contained in the initial distribution of transverse relaxation time (T ₂) data of fresh A. bisporus slices: T ₂₁ (0.38–7.05 ms), T ₂₂ (9.33–32.75 ms), T ₂₃₁ (37.65–265.61 ms), and T ₂₃₂ (305.39–811.13 ms). Thermosonic pretreatment significantly decreased the initial free water content of A. bisporus sample but was accompanied by a sharp increase in its immobilized water. “Semi-bound water transfer” appeared during microwave vacuum drying (MVD) at moisture contents (X _w ) of 0.70 and 0.60 g/g (wet basis (w.b.)) for untreated and thermosonically treated samples, respectively. MVD caused dramatic changes in the water state and enhanced the T _g by decreasing the content and mobility of immobilized water in A. bisporus tissues. The mobility of semi-bound water for thermosonically and MVD-treated samples was higher than for MVD-untreated samples, resulting in T _g values decreasing by approximately 2–11.5 °C, but the uniformity of water distribution in thermosonic-treated and MVD-treated samples was better at X _w  ≤ 0.52 g/g (w.b.).     

Application of Bismuth(III)-Entrapped XO Biosensing System for Xanthine Determination in Beverages

A xanthine biosensor was prepared by electrochemical immobilization of xanthine oxidize enzyme onto carbon paste electrode via entrapment of Bi³⁺. After the optimization of experimental parameters, analytical characteristics were investigated. Two linear ranges between 0.02 and 0.06 and 1–7.5 μM with the equation y?=?93.00x?+?0.12 and y?=?1.07x?+?18.03 with the correlation coefficients of R ²?=?0.9951 and R ²?=?0.9931, respectively, were obtained for this biosensing system. RSD value was calculated for 0.04 μM xanthine (n?=?5) and found as 3.84%. LOD and LOQ values were also calculated and revealed as 1.30?×?10^?8 and 4.3?×?10^?8 M, respectively. Then, this biosensor was applied for xanthine detection in real samples. As a sample treatment, only necessary dilutions were made. Four types of beverages including wine, energy drink, peach, and sour cherry juice were used for this purpose. Obtained recovery values demonstrate that this system is applicable for xanthine detection in real samples without needing any laborious sample pretreatment procedures.     

Combined Application of Fluorescence Spectroscopy and Chemometrics Analysis in Oxidative Deterioration of Edible Oils

Combined methods of fluorescence spectrometry with chemometrics were used to monitor oxidation deterioration of edible oil. Synchronous and three dimensional fluorescence spectroscopy techniques were proposed for monitoring palm oil, camellia oil, sunflower oil and perilla oil during oven accelerated oxidation. Principal component analysis plot of fluorescence intensity (λex = 320–700 nm) clearly showed oxidative evolution of oils over heating time. High saturated or monounsaturated oils exhibited high regression coefficients between peroxide values and fluorescence intensity (R ²  = 0.973 for 400 nm in palm oil; R ²  = 0.956 for 370 nm in camellia oil). High diunsaturated oil exhibited high regression coefficient between nonpolar carbonyl compounds and fluorescence intensity (R ²  = 0.970 for 370 nm in sunflower oil). High triunsaturated oil exhibited high regression coefficient between p-anisidine value and fluorescence intensity (R ²  = 0.938 for 665 nm in perilla oil). In conclusion, Fluorescence spectroscopy is a rapid and green nondestructive method for oxidation monitoring. Differences of fatty acid compositions played key rules in formation of oxidation products and evolution of fluorescence spectra.     

Evaluation of <Emphasis Type="Italic">eryC</Emphasis> as a Molecular Marker for the Quantitative Detection of <Emphasis Type="Italic">Brucella</Emphasis> Spp. by Real-Time PCR in Food Samples

We evaluated the capacity of the Brucella sp. eryC gene as a diagnostic marker for brucellosis by quantitative real-time PCR. eryC gene encodes the enzyme d-erythrulose-1-phosphate dehydrogenase that plays an important role in the erythritol metabolism and is related with the Brucella survival in the intracellular environment of the macrophage. The assay includes an internal amplification control (IAC) in order to avoid false negative results. It was 100% specific, with an analytical sensitivity of 1 genome equivalent (GE) in 43% of the reactions, being the quantification highly linear (R ² > 0.9953) and efficient (PCR efficiency >0.8820) over a 6-log dynamic range, down to 10 GE. Finally, the applicability of this assay was evaluated with artificially contaminated biological matrices implicated in the transmission of this bacterium such as sheep raw milk and pig blood. The eryC-IAC real-time PCR assay allowed detection of as few as ten Brucella cells per 25 ml of sheep raw milk or per 1 ml of pig blood. In conclusion, we present an alternative for the detection of Brucella genus and therefore facilitate the establishment of preventive and prophylactic measures in food and farm environments.