Mass spectrometry in food authentication and origin traceability.

Food authentication and origin traceability are popular research topics, especially as concerns about food quality continue to increase. Mass spectrometry (MS) plays an indispensable role in food authentication and origin traceability. In this review, the applications of MS in food authentication and origin traceability by analyzing the main components and chemical fingerprints or profiles are summarized. In addition, the characteristic markers for food authentication are also reviewed, and the advantages and disadvantages of MS-based techniques for food authentication, as well as the current trends and challenges, are discussed. The fingerprinting and profiling methods, in combination with multivariate statistical analysis, are more suitable for the authentication of high-value foods, while characteristic marker-based methods are more suitable for adulteration detection. Several new techniques have been introduced to the field, such as proton transfer reaction mass spectrometry, ambient ionization mass spectrometry (AIMS), and ion mobility mass spectrometry, for the determination of food adulteration due to their fast and convenient analysis. As an important trend, the miniaturization of MS offers advantages, such as small and portable instrumentation and fast and nondestructive analysis. Moreover, many applications in food authentication are using AIMS, which can help food authentication in food inspection/field analysis. This review provides a reference and guide for food authentication and traceability based on MS.

Systematic analysis and expression of Gossypium ATG8 family reveals the roles of GhATG8f responding to salt stress in cotton.

KEY MESSAGE: Comprehensive analysis of Gossypium ATG8 family indicates that GhATG8f could improve salt tolerance of cotton by increasing SOD, POD and CAT activity and proline accumulation. In plants, autophagy is regulated by several genes that play important roles in initiating and controlling the process. ATG8, functioning as a protein similar to ubiquitin, is involved in crucial tasks throughout the autophagosome formation process. In this research, we conducted an extensive and all-encompassing investigation of 64 ATG8 genes across four varieties of cotton. According to the subcellular localization prediction results, 49 genes were found in the cytoplasm, 6 genes in the chloroplast, 1 gene in the peroxisome, 5 genes in the nucleus, and 3 genes in the extracellular region. Phylogenetic analysis categorized a total of 5 subfamilies containing sixty-four ATG8 genes. The expression of the majority of GhATG8 genes was induced by salt, drought, cold, and heat stresses, as revealed by RNA-seq and real-time PCR. Analysis of cis-elements in the promoters of GhATG8 genes revealed the predominant presence of responsive elements for plant hormones and abiotic stress, suggesting that GhATG8 genes might have significant functions in abiotic stress response. Furthermore, we additionally performed a gene interaction network analysis for the GhATG8 proteins. The salt stress resistance of cotton was reduced due to the downregulation of GhATG8f expression, resulting in decreased activity of CAT, SOD, and POD enzymes, as well as decreased fresh weight and proline accumulation. In summary, our research is the initial exploration of ATG8 gene components in cotton, providing a basis for future investigations into the regulatory mechanisms of ATG8 genes in autophagy and their response to abiotic stress.

Effects of UV-B Radiation Exposure on Transgenerational Plasticity in Grain Morphology and Proanthocyanidin Content in Yuanyang Red Rice.

The effect of UV-B radiation exposure on transgenerational plasticity, the phenomenon whereby the parental environment influences both the parent's and the offspring's phenotype, is poorly understood. To investigate the impact of exposing successive generations of rice plants to UV-B radiation on seed morphology and proanthocyanidin content, the local traditional rice variety 'Baijiaolaojing' was planted on terraces in Yuanyang county and subjected to enhanced UV-B radiation treatments. The radiation intensity that caused the maximum phenotypic plasticity (7.5 kJ·m-2) was selected for further study, and the rice crops were cultivated for four successive generations. The results show that in the same generation, enhanced UV-B radiation resulted in significant decreases in grain length, grain width, spike weight, and thousand-grain weight, as well as significant increases in empty grain percentage and proanthocyanidin content, compared with crops grown under natural light conditions. Proanthocyanidin content increased as the number of generations of rice exposed to radiation increased, but in generation G3, it decreased, along with the empty grain ratio. At the same time, biomass, tiller number, and thousand-grain weight increased, and rice growth returned to control levels. When the offspring's radiation memory and growth environment did not match, rice growth was negatively affected, and seed proanthocyanidin content was increased to maintain seed activity. The correlation analysis results show that phenylalanine ammonialyase (PAL), cinnamate-4-hydroxylase (C4H), dihydroflavonol 4-reductase (DFR), and 4-coumarate:CoA ligase (4CL) enzyme activity positively influenced proanthocyanidin content. Overall, UV-B radiation affected transgenerational plasticity in seed morphology and proanthocyanidin content, showing that rice was able to adapt to this stressor if previous generations had been continuously exposed to treatment.

Effects and Mechanism of Enhanced UV-B Radiation on the Flag Leaf Angle of Rice.

Leaf angle is an influential agricultural trait that influences rice (Oryza sativa L.) plant type and yield, which results from the leaf bending from the vertical axis to the abaxial axis. UV-B radiation affects plant morphology, but the effects of varying UV-B intensities on rice flag leaves and the underlying molecular, cellular, and physiological mechanisms remain unknown. This experiment aims to examine the effect of natural light and field-enhanced UV-B radiation (2.5, 5.0, 7.5 kJ·m-2) on the leaf angle of the traditional rice variety Baijiaolaojing on Yuanyang terraces. In comparison with natural light, the content of brassinolide and gibberellin in rice flag leaves increased by 29.94% and 60.1%, respectively. The auxin content decreased by 17.3%. Compared with the natural light treatment, the cellulose content in the pulvini was reduced by 13.8% and hemicellulose content by 25.7% under 7.5 kJ·m-2 radiation intensity. The thick-walled cell area and vascular bundle area of the leaf pulvini decreased with increasing radiation intensity, and the growth of mechanical tissue in the rice leaf pulvini was inhibited. The flag leaf angle of rice was greatest at 7.5 kJ·m-2 radiation intensity, with an increase of 50.2%. There are two pathways by which the angle of rice flag leaves is controlled under high-intensity UV-B radiation. The leaf angle regulation genes OsBUL1, OsGSR1, and OsARF19 control hormone levels, whereas the ILA1 gene controls fiber levels. Therefore, as cellulose, hemicellulose, sclerenchyma, and vascular bundles weaken the mechanical support of the pulvini, the angle of the flag leaf increases.

Extraction and Determination of Vitamin K1 in Foods by Ultrasound-Assisted Extraction, SPE, and LC-MS/MS.

Vitamin K1 is one of the important hydrophobic vitamins in fat-containing foods. Traditionally, lipase is employed in the determination of vitamin K1 to remove the lipids, which makes the detection complex, time-consuming, and insensitive. In this study, the determination of vitamin K1 in fat-containing foods was developed based on ultrasound-assisted extraction (UAE), solid-phase extraction (SPE) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). The optimal conditions for extraction of vitamin K1 were material-liquid ratio of 1:70 (g/mL), extraction temperature of 50 °C, extraction power of 700 W, extraction time of 50 min, material-wash fluid ratio of 1:60 (g/mL), and 8 mL of hexane/anhydrous ether (97:3, v/v) as the elution solvent. Then, vitamin K1 was analyzed on a ZORBAX SB-C18 column (50 mm × 2.1 mm, 1.8 µm) by gradient elution with water (0.01% formic acid) and methanol (0.01 formic acid + 2.5 mmol/L ammonium formate) as the mobile phase. The limit of detection (LOD) and limit of quantification (LOQ) were 0.05 and 0.16 µg/kg, respectively. Calibration curve was linear over the range of 10-500 ng/mL (R2 > 0.9988). The recoveries at three spiked levels were between 80.9% and 119.1%. The validation and application indicated that the proposed method was simple and sensitive in determination of vitamin K1 in fat-containing foods.

Monitoring Metabolite Production of Aflatoxin Biosynthesis by Orbitrap Fusion Mass Spectrometry and a D-Optimal Mixture Design Method.

Aflatoxins, highly toxic and carcinogenic to humans, are synthesized via multiple intermediates by a complex pathway in several Aspergilli, including Aspergillus flavus. Few analytical methods are available for monitoring the changes in metabolite profiles of the aflatoxin biosynthesis pathway under different growth and environmental conditions. In the present study, we developed by a D-optimal mixture design a solvent system, methanol/dichloromethane/ethyl acetate/formic acid (0.36/0.31/0.32/0.01), that was suitable for extracting the pathway metabolites. The matrix effect from dilution of cell extracts was negligible. To facilitate the identification of these metabolites, we constructed a fragmentation ion library. We further employed liquid chromatography coupled with high-resolution mass spectroscopy (UHPLC-HRMS) for simultaneous quantification of the metabolites. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.002-0.016 and 0.008-0.05 µg/kg, respectively. The spiked recovery rates ranged from 81.3 to 100.3% with intraday and interday precision less than 7.6%. Using the method developed to investigate the time-course aflatoxin biosynthesis, we found that precursors, including several possible toxins (with a carcinogenic group similar to aflatoxin B1), occurred together with aflatoxin, and that production increased rapidly at the early growth stage, peaked on day four, and then decreased substantially. The maximum production of aflatoxin B1 and aflatoxin B2 occurred 1 day later. Moreover, the dominant branch pathway was the one for aflatoxin B1 formation. We revealed that the antiaflatoxigenicity mechanism of Leclercia adecarboxylata WT16 was associated with a factor upstream of the aflatoxin biosynthesis pathway. The design strategies can be applied to characterize or detect other secondary metabolites to provide a snapshot of the dynamic changes during their biosynthesis.

Multispecies Adulteration Detection of Camellia Oil by Chemical Markers.

Adulteration of edible oils has attracted attention from more researchers and consumers in recent years. Complex multispecies adulteration is a commonly used strategy to mask the traditional adulteration detection methods. Most of the researchers were only concerned about single targeted adulterants, however, it was difficult to identify complex multispecies adulteration or untargeted adulterants. To detect adulteration of edible oil, identification of characteristic markers of adulterants was proposed to be an effective method, which could provide a solution for multispecies adulteration detection. In this study, a simple method of multispecies adulteration detection for camellia oil (adulterated with soybean oil, peanut oil, rapeseed oil) was developed by quantifying chemical markers including four isoflavones, trans-resveratrol and sinapic acid, which used liquid chromatography tandem mass spectrometry (LC-MS/MS) combined with solid phase extraction (SPE). In commercial camellia oil, only two of them were detected of daidzin with the average content of 0.06 ng/g while other markers were absent. The developed method was highly sensitive as the limits of detection (LODs) ranged from 0.02 ng/mL to 0.16 ng/mL and the mean recoveries ranged from 79.7% to 113.5%, indicating that this method was reliable to detect potential characteristic markers in edible oils. Six target compounds for pure camellia oils, soybean oils, peanut oils and rapeseed oils had been analyzed to get the results. The validation results indicated that this simple and rapid method was successfully employed to determine multispecies adulteration of camellia oil adulterated with soybean, peanut and rapeseed oils.

Identification of Nutritional Components in Black Sesame Determined by Widely Targeted Metabolomics and Traditional Chinese Medicines.

Chemical composition of secondary metabolites is of great importance for quality control of agricultural products. Black sesame seeds are significantly more expensive than white sesame seeds, because it is thought that black sesame seeds are more beneficial to human health than white sesame seeds. However, the differences in nutrient composition between black sesame seeds and white sesame seeds are still unknown. The current study examined the levels of different metabolites in black and white sesame seeds via the use of a novel metabolomics strategy. Using widely targeted metabolomics data, we obtained the structure and content of 557 metabolites, out of which 217 metabolites were identified, and discovered 30 metabolic pathways activated by the secondary metabolites in both black and white sesame seeds. Our results demonstrated that the main pathways that were differentially activated included: phenylpropanoid biosynthesis, tyrosine metabolism, and riboflavin metabolism. More importantly, the biomarkers that were significantly different between black seeds and white sesame seeds are highly related to the functions recorded in traditional Chinese medicine. The results of this study may serve as a new theoretical reference for breeding experts to promote the genetic improvement of sesame seeds, and therefore the cultivation of higher quality sesame varieties.

Discrimination between the Triglyceride Form and the Ethyl Ester Form of Fish Oil Using Chromatography-Mass Spectrometry.

Although the triglyceride form is the natural form of fish oil found in fish, the ethyl ester form of fish oil, which is used during processing to save costs, is also present on the market. In this study, fatty acids and lipids were determined using gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-linear ion trap mass spectrometry (LC-LIT/MS), respectively, according to developed methods. The identification of fatty acids was based on the mass spectral characteristics and equivalent chain lengths. However, the fatty acid contents of both forms of fish oils are quite similar. The application of the LC-LIT/MS method for the structural characterization of triacylglycerols (TAGs) and the mechanism of LIT/MS fragmentation are also discussed. Neutral losses of CH2=CH2 (m/z 28) and CH3CH2OH (m/z 46), which are LIT/MS characteristics of ethyl ester from fish oil, were found for the first time. The triglyceride form of fish oils was easily and accurately identified using fingerprint chromatography. In conclusion, lipid analysis combined with LC-LIT/MS showed an improved capability to distinguish between types of fish oil.

Single extraction and integrated non-target data acquisition with data mining workflow for analysis of hazardous substances in agricultural plant products.

Identifying contaminants in agricultural plant food products (APFPs) is a major problem. In this study, we developed a single-step extraction and integrated non-target data acquisition (INDA) workflow for increasing hazardous substances coverage. D-optimal experimental designs were applied to optimize filter plate extraction (FPE) for one-single extraction of multipolar hazardous substances. The vDIA mode was used to collect all precursor ion fragments within the range to supplement data loss caused by DDA mode. The underlying principle of vDIA is to increase the utilization rate of MS2 spectra that are likely to identify a maximum number and minimum amounts of hazardous substances. Compared with traditional DDA mode alone, a combination of the two modes increased the rate of identification of hazardous substances by 18.5%. The molecular network of hazardous substance provided by GNPS could enable some metabolites and structure-related products to discover potentially hazardous substance.

Screening for pesticide residues in oil seeds using solid-phase dispersion extraction and comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry.

In this paper, we describe the development of an oil-absorbing matrix solid-phase dispersion extraction with comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry suitable for screening of 68 pesticide residues (PRs) in peanut, soybean, rape seed, sesame, and sunflower seed. The 68 PRs include 27 kinds of organophosphorus, 23 organic chlorines, 11 synthetic pyrethroids, and 7 carbamates. Heptachlor epoxide was used as the internal standard. Aminopropyl silica was chosen as the dispersion sorbent of the oil-absorbing matrix solid-phase dispersion extraction and was applied to capture hydrophobic components from high oil samples. A 35-min orthogonal separation was performed by using comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry with a nonpolar-polar column set. Identification of 68 PRs in the extract was finished by using the time-of-flight mass spectrometry in the assistance of an automated peak-find and spectral deconvolution software. A screening based on control design was introduced and explained. This screening method considerably reduced the cost for the quantitative and confirmatory analyses. The quality of present screening method was evaluated by the Document No. SANCO/10684/2009. The false positive rate and false negative rate provide a useful tool for the evaluation of screening performance.

Identification of glycerophospholipids using self-built recognition software based on positive and negative ion high-resolution mass spectrometric fragmentation experiments.

Glycerophospholipids (GPs) have a wide variety and complex structure, which makes their identification challenging. Our software affords a novel tool for the automated identification of non-target GPs in biological mixtures. Here, we explored the multi-stage fragmentation processes of GPs in positive and negative ion modes, and then constructed multi-stage fragment ion databases. This database includes 8214 simulated GP molecules from a random combination of fatty acids corresponding to 42,439 self-built predicted multi-stage fragment ions in positive ion mode and 31,487 self-built predicted multi-stage fragment ions in negative ion mode (MS ≤ 3). The automatic GP identification (AGPI) software can screen out GP candidates utilizing the MS1 accurate mass. The isomers of fatty acid chains and the phosphoryl head group can be distinguished using the MS2 and MS3 fragment spectra in positive-ion and negative-ion modes. All of the selected 45 GP standards were putatively identified using AGPI software; however, there were false positives because the software cannot distinguish positional isomers of fatty acids. Therefore, the AGPI software could be applied to identify GPs in samples, such as cancer cells; we successfully identified 41 GPs in cancer cells.

Quantitative analysis of metabolites in the aflatoxin biosynthesis pathway for early warning of aflatoxin contamination by UHPLC-HRMS combined with QAMS.

Aflatoxins seriously threaten human health and food safety, and early warning benefits the reasonable use of control measures to reduce aflatoxin contamination. In this study, a novel method for quantifying aflatoxins and their precursors in the aflatoxin biosynthesis pathway was developed by combining ultra-high performance liquid chromatography high-resolution mass spectrometry (UHPLC-HRMS) with quantitative analysis of multi-components by a single marker (QAMS). The stability of the relative correction factor (RCF) of QAMS was then systematically evaluated. The validation results showed that the relative deviation (RD) between QAMS and the external standard method (ESM) was less than 11.7%, indicating that the established QAMS method could replace ESM without the use of reference standards. This method was successfully employed to compare the time-course changes of metabolites in the aflatoxin biosynthesis pathway of Aspergillus flavus and Aspergillus parasitica. As a result, the precursors of (1'S,5'R)-5'-Hydroxyaverantin (HAVN) and Versicolorin B (VerB) could be used as potential markers for the early warning of aflatoxin contamination. This study provided a quantitative method of aflatoxins and their precursors in the biosynthesis pathway, and may serve as a reference for the extension of quantitative studies on other metabolic pathways.

Fungi population metabolomics and molecular network study reveal novel biomarkers for early detection of aflatoxigenic Aspergillus species.

Mycotoxins threaten global food safety, public health and cause huge socioeconomic losses. Early detection is an effective preventive strategy, yet efficient biomarkers for early detection of aflatoxigenic Aspergillus species are lacking. Here, we proposed to use untargeted metabolomics and machine learning to mine biomarkers of aflatoxigenic Aspergillus species. We systematically delineated metabolic differences across 568 extensive field sampling A. flavus and performed biomarker analysis. Versicolorin B, 11-hydroxy-O-methylsterigmatocystin et.al metabolites shown a high correlation (from 0.71 to 0.95) with strains aflatoxin-producing capacity. Molecular networking analysis deciphered the connection of aflatoxins and biomarkers as well as potential emerging mycotoxins. We then developed a model using the biomarkers as variables to discern aflatoxigenic Aspergillus species with 97.8% accuracy. A validation dataset and metabolome from other 16 fungal isolates confirmed the robustness and specificity of these biomarkers. We further demonstrated the solution feasibility in agricultural products by early detection of biomarkers, which predicted aflatoxin contamination risk 35-47 days in advance. A developed operable decision rule by the XGBoost algorithm help regulators to intuitively assess the risk prioritization with 87.2% accuracy. Our research provides novel insights into global food safety risk assessment which will be crucial for early prevention and control of mycotoxins.

Adulteration detection of essence in sesame oil based on headspace gas chromatography-ion mobility spectrometry.

Sesame oil is a traditional and delicious edible oil in China and Southeast Asia with a high price. However, sesame oil essence was often illegally added to cheaper edible oils to counterfeit sesame oil. In this study, a rapid and accurate headspace gas chromatography-ion mobility spectrometry (HS-GC-IMS) method was proposed to detect the counterfeit sesame oil where the other cheap oils were adulterated with essence. Combined with chemometric methods including principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA) and random forest (RF), authentic and counterfeit sesame oils adulterated with sesame essence (0.5%, w/w) were easily separated into two groups. More importantly, 2-methylbutanoic acid, 2-furfurylthiol, methylpyrazine, methional, and 2,5-dimethylpyrazine were found to be markers of sesame essence, which were used to directly identify the sesame essence. The determination of volatile compounds based on HS-GC-IMS was proven to be an effective method for adulteration detection of essence in sesame oil.

Simultaneous determination of 13 phytohormones in oilseed rape tissues by liquid chromatography-electrospray tandem mass spectrometry and the evaluation of the matrix effect.

In the experiment, a high-performance liquid chromatography and electrospray ionization-tandem mass spectrometry with selected reaction monitoring was used to simultaneously determine various classes of phytohormones, including indole-3-acetic acid, α-naphthaleneacetic acid, 2-chlorobenzoic acid, 4-chlorobenzoic acid, indole-3-butyric acid, gibberellic acid, 2,4-dichlorophenoxyacetic acid, 2-naphthoxyacetic acid, abscisic acid, 2,3,5-triiodobenzoic acid, uniconazole, paclobutrazol and 2,4-epibassinolide in rape tissues. The analyses were separated by an HPLC equipped with a reversed-phase column using a binary solvent system composed of methanol and water, both containing 0.1% of formic acid. The matrix effect was also considered and determined. The technology was applied to analyze rape tissues, including roots, stems, leaves, flowers, immature pods and rape seeds. The rape tissues were subjected to ultrasound-assisted extraction and purified by dispersive solid-phase extraction, and then transferred into the liquid chromatography system. The detection limit for each plant hormone was defined by the ratio of signal/background noise (S/N) of 3. The results showed perfect linearity (R(2) values of 0.9987-1.0000) and reproducibility of elution times (relative standard deviations, RSDs,<1%) and peak areas (RSDs,<7%) for all target compounds.

[Coinfection effects of porcine circovirus type 2 and porcine parvovirus in vivo on phagocytosis and interferon mRNA expression of porcine alveolar macrophages].

OBJECTIVE: Our study is to analyse the coinfection effects of porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV) in vivo on phagocytosis and interferon mRNA expression level of porcine alveolar macrophages (PAM). METHODS: Forty-eight 5-week-old healthy piglets were divided randomly into 4 groups of 12 (PCV2, PPV, PCV2/PPV and the control groups). The piglets in PCV2 group were inoculated oronasally with 3 mL of porcine circovirus type2 (PCV2, 10(5.61) TCID50/0.1 mL), PPV group with 3 mL of porcine parvovirus (PPV, 10(6.69) TCID50/0.1 mL), PCV2/PPV group with 3 mL of PCV2 and 3 mL of PPV and the control group with 3 mL of cell culture medium, respectively. Three piglets from each group were sacrificed randomly on 3, 7, 14 and 35 day post infection (dpi) and porcine alveolar macrophages (PAM) were collected to detect viability, phagocytotic capabilities and alpha1- and gamma-interferon (IFN-alpha1 and IFN-gamma) mRNA levels of PAM. RESULTS: The viabilities of PAM from PCV2 group and PCV2/PPV group became weaker than that of control group during the period of 3 - 14 dpi but they were similar to that of control group on 35 dpi; there was no significant difference between the viability of PCV2/PPV group and that of PCV2 group (P > 0.05). The phagocytotic capabilities of PAM from three virus infection groups were lower than that of control group (P < 0.01), among which that of PCV2/PPV group descended more drastically. IFN-alpha1 and IFN-gamma mRNA levels in PAM from PCV2/PPV group were significantly lower than those of PCV2, PPV and control groups (P < 0.01). CONCLUSION: PCV2/PPV co-infection did not cause further decline of PAM viability but strongly weakened phagocytosis and constantly lowered IFN (IFN-alpha1 and IFN-gamma) mRNA expression levels of PAM.

UHPLC-HRMS-Based Untargeted Lipidomics Reveal Mechanism of Antifungal Activity of Carvacrol against Aspergillus flavus.

Aspergillus flavus is a common contaminant in grain, oil and their products. Its metabolite aflatoxin B1 (AFB1) has been proved to be highly carcinogenic. Therefore, it is of great importance to find possible antifungal substances to inhibit the growth and toxin production of Aspergillus flavus. Carvacrol (CV) was reported as a potent antifungal monoterpene derived from plants. In this paper, the antifungal effects and mechanism of CV on Aspergillus flavus were investigated. CV was shown good inhibition on the growth of Aspergillus flavus and the production of AFB1. CV used in concentrations ranging from 0, 50, 100 and 200 µg/mL inhibited the germination of spores, mycelia growth and AFB1 production dose-dependently. To explore the antifungal mechanism of CV on Aspergillus flavus, we also detected the ergosterol content of Aspergillus flavus mycelia, employed Scanning Electron Microscopy (SEM) to observe mycelia morphology and utilized Ultra-High-Performance Liquid Chromatography-High-Resolution Mass Spectrometry (UHPLC-HRMS) to explore the lipidome profiles of Aspergillus flavus. The results showed that the production of ergosterol of mycelia was reduced as the CV treatment concentration increased. SEM photographs demonstrated a rough surface and a reduction in the thickness of hyphae in Aspergillus flavus treated with CV (200 µg/mL). In positive ion mode, 21 lipids of Aspergillus flavus mycelium were downregulated, and 11 lipids were upregulated after treatment with 200-µg/mL CV. In negative ion mode, nine lipids of Aspergillus flavus mycelium were downregulated, and seven lipids upregulated after treatment with 200-µg/mL CV. In addition, the analysis of different lipid metabolic pathways between the control and 200-µg/mL CV-treated groups demonstrated that glycerophospholipid metabolism was the most enriched pathway related to CV treatment.

Identification and Validation of Metabolic Markers for Adulteration Detection of Edible Oils Using Metabolic Networks.

Food adulteration is a challenge faced by consumers and researchers. Due to DNA fragmentation during oil processing, it is necessary to discover metabolic markers alternative to DNA for adulteration detection of edible oils. However, the contents of metabolic markers vary in response to various factors, such as plant species, varieties, geographical origin, climate, and cultivation measures. Thus, it is difficult to identify a universal marker for all adulterants that may be present in some authentic samples. Currently, the specificity and selectivity of metabolic biomarkers are difficult to validate. Therefore, this study developed a screening strategy based on plant metabolic networks by developing a targeted analytical method for 56 metabolites in a metabolic network, using liquid/liquid extraction-liquid chromatography-tandem mass spectrometry (LC-MS/MS). We identified a chain of 11 metabolites that were related to isoflavonoid biosynthesis, which were detected in soybean oils but not rapeseed oils. Through multiple-marker mutual validation, these metabolites can be used as species-specific universal markers to differentiate soybean oil from rapeseed oil. Moreover, this method provides a model for screening characteristic markers of other edible vegetable oils and foods.

Determination of pyrethroids in porcine tissues by matrix solid-phase dispersion extraction and high-performance liquid chromatography.

An effective matrix solid-phase dispersion (MSPD) extraction for determination of two pyrethroids (cypermethrin and deltamethrin) in porcine tissues (liver, muscle, heart and kidney) is described. A neutral alumina-based MPSD column was used for extraction of analytes. The high-performance liquid chromatography and ultraviolet detector was applied using a reverse-phase C(18) column and acetonitrile-water (85:15, v/v) was used as mobile phase. The good linear fit curve ranging from 0.05 to 50µgmL(-1) for cypermethrin (CM) and deltamethrin (DM) was obtained with a regression coefficient (r) of 0.999. Recoveries at 0.2 and 0.5µgg(-1) levels were between 83.5% and 109%. The limits of detection and quantification were: 0.01 and 0.026µgg(-1) for CM, 0.017 and 0.056µgg(-1) for DM, respectively. The proposed method was successfully applied to the determination of the pyrethroids in porcine tissues.