Assessment of the Adverse Health Effects of Aflatoxin Exposure from Unpackaged Peanut Oil in Guangdong, China.

Aflatoxins are liver carcinogens and are common contaminants in unpackaged peanut (UPP) oil. However, the health risks associated with consuming aflatoxins in UPP oil remain unclear. In this study, aflatoxin contamination in 143 UPP oil samples from Guangdong Province were assessed via liquid chromatography-tandem mass spectrometry (LC-MS). We also recruited 168 human subjects, who consumed this oil, to measure their liver functions and lipid metabolism status. Aflatoxin B1 (AFB1) was detected in 79.72% of the UPP oil samples, with levels ranging from 0.02 to 174.13 µg/kg. The average daily human intake of AFB1 from UPP oil was 3.14 ng/kg·bw/day; therefore, the incidence of liver cancer, caused by intake of 1 ng/kg·bw/day AFB1, was estimated to be 5.32 cases out of every 100,000 persons per year. Meanwhile, Hepatitis B virus (HBV) infection and AFB1 exposure exerted a synergistic effect to cause liver dysfunction. In addition, the triglycerides (TG) abnormal rate was statistically significant when using AFB1 to estimate daily intake (EDI) quartile spacing grouping (p = 0.011). In conclusion, high aflatoxin exposure may exacerbate the harmful effects of HBV infection on liver function. Contamination of UPP oil with aflatoxins in Guangdong urgently requires more attention, and public health management of the consumer population is urgently required.

Global systematic review and meta-analysis on prevalence and concentration of aflatoxins in peanuts oil and probabilistic risk assessment.

Exposure to mycotoxins in food is largely unavoidable, and concerns about their health effects are growing. Consumption of vegetable oils such as peanuts oil has increased, hence several studies have been conducted on concentration of aflatoxins (AFs) in peanuts oil. Search was performed in Scopus and PubMed databases on prevalence and concentration of AFs in peanuts oil from 1 January 2005 to 15 April 29, 2022. Prevalence and concentration of AFs in peanuts oil was meta-analyzed based on country and type of AFs subgroups. In addition, health risk was calculated using monte carlo simulation method. Pooled prevalence of AFB1 in peanuts oil was 47.9%; AFB2, 46.45%; AFG1, 46.92% and AFG2, 54.01%. The Overall prevalence of AFTs was 49.30%, 95%CI (35.80-62.84%). Pooled concentration of AFB1 in peanuts oil was 2.30 µg/kg; AFB2, 0.77 µg/kg; AFG1, 0.07 µg/kg; AFG1, 0.28 µg/kg. The sort of country based on mean of MOEs in the adults consumers was Japan (47,059) > China (17,670) > Ethiopia (7,398) > Sudan (6,974) > USA (1,012) and sort of country based on mean of MOEs in the children was Japan (120,994) > China (46,991) > Ethiopia (19,251) > Sudan (18,200) > USA (2,620). Therefore, adults consumers were in considerable health risk in Ethiopia, Sudan and USA and for children in USA (MOE < 10,000).

Occurrence and exposure assessment of aflatoxins in Zhejiang province, China.

The purpose of this study was to assess the risk of aflatoxins due to multiple food consumption among the Zhejiang population. Ultra-high-performance liquid chromatography coupled with tandem mass spectrometry method was used to determine aflatoxins in 792 samples. Aflatoxins were detected in 27.1% of the samples at levels between 0.07 and 262.63 µg kg-1, and aflatoxins B1 was the most frequently detected among different types of samples. 0.8% of peanut oil, 3.39% of nut products as well as 1.1% of condiments contaminated with aflatoxins B1 exceeded China national tolerance limits. Peanut oil had the highest incidence of aflatoxin, with a range from 0.17 to 22.50 µg kg-1. Using bags conferred limited advantages in reducing aflatoxin contents. Moreover, peanut and rice were the main contributors to dietary exposure to aflatoxins among Zhejiang residents. Finally, the margin of exposure values obtained by rice consumption were far from the safe margin of 10,000, indicating a potential risk to public health. The results pointed out the need for further prioritization of aflatoxins B1 risk-management actions in Zhejiang.

Effect of Drying Methods on Peanut Quality during Storage.

Storage is an important step after peanut harvest and drying. Many factors could affect the peanut quality during storage. The quality change differences of peanut after being dried by solar radiation and at 35°C, 40°C, 45°C, 50°C during later storage were investigated, including moisture content (MC) and germination percentage (GP) of peanut kernels, acid value (AV), peroxide value (PV), iodine value (IV), vitamin E (VE) content and fatty acid composition (FAC) of extracted peanut oil. And the impact of four storage conditions, air-room temperature (A-RT), air-low temperature (A-LT), vacuum-room temperature (V-RT) and nitrogen-room temperature (N-RT) on peanut quality after 10 months' storage were also studied in this paper. The results revealed that drying conditions had only a little influence on peanut quality during later storage. Peanut dried by solar radiation was more easily oxidized than that dried under other drying conditions. The effects of storage time were much greater. The GP, AV, PV, VE content and FAC, showed significantly changes along with storage. GP and VE content decreased, AV and PV increased, and some linoleic acid was oxidized to oleic acid after 10 months' storage. In addition, A-LT exhibited best performance in keeping peanut quality than A-RT, V-RT and N-RT, which demonstrated that low temperature was more advantageous for peanut storage than controlled atmosphere. These results above would provide useful information and reference for the peanut storage to apply in food industry.

Peanut selenium distribution, concentration, speciation, and effects on proteins after exogenous selenium biofortification.

Fortification of Se is vital importance for both nutritional demand and prevention of Se-deficiency-related diseases. To better understand t selenium distribution, concentration, speciation, its effects on proteins, and cytotoxic activity, the biofortification of exogenous Se in peanut was conducted in this study. Our data have shown that foliar spraying of Se-riched fertilizer was more efficient for biotransformation of inorganic Se to organic Se by peanut plant. Besides, the Se content in peanut was increased in a dose-dependent manner. Our present study also confirmed that SeCys2, MeSeCys, and SeMet were the main Se speciation within peanut proteins. Moreover, the secondary structure and thermostability of peanut protein were altered as a result of the Se treatments, and these alterations could be attributed to the replacements of cysteine and methionine by selenocysteine and selenomethionine, respectively. The Se-enriched peanut protein could significantly inhibit the growth of Caco-2 and HepG2 in a concentration-dependent manner.

Effects of Pretreatment on the Yield of Peanut Oil and Protein Extracted by Aqueous Enzymatic Extraction and the Characteristics of the Emulsion.

Effects of comminution on peanut particle size and yield of peanut oil and protein were analyzed. Additionally, the emulsion properties (surface protein concentration, particle size, and ξ-potential) were compared. Moreover, different demulsification methods were used to investigate the emulsion stability. Results showed that the yield of peanut oil and protein was highest (87.23% and 82.05%, respectively) after dry comminution for 72 s. Upon wet comminution for 120 s, the yields of peanut oil and protein were 89.91% and 84.70%, respectively, which were both significantly higher than that obtained after dry comminution (p < 0.05). The surface protein concentration and ξ-potential of emulsion made by dry comminution (DCE) were 7.02 mg/m2 and 12.08 mV, respectively, and those of emulsion made by wet comminution (WCE) were 10.71 mg/m2 and 15.25 mV, respectively, which were significantly higher than that of DCE (p < 0.05). The volume average particle size of DCE was 3.41 µm, which was significantly higher than that of WCE (3.18 µm, p < 0.05). Collectively, these results indicated that WCE was more stable than DCE. Further, the demulsification rate of DCE was significantly higher than that of WCE when treated by freeze-thawing, pH, papain, and phospholipase A2 (p < 0.05). Demulsification effect of Alcalase 2.4L was the best among these five demulsification methods treated, and the demulsification rate of DCE reached 92.77%, which was slightly higher than that of WCE (92.67%), further illustrating the higher stability of WCE.

Fate of phospholipids during aqueous extraction processing of peanut and effect of demulsification treatments on oil-phosphorus-content.

PC (phosphatidylcholine), PE (phosphatidylethanolamine), PI (phosphatidylinositol), and PA (phosphatidic acid) in 9 peanut matrices obtained during the AEP (aqueous extraction processing) of peanut were quantified employing HPLC-ELSD analysis in this study. Phosphorus contents of crude oils obtained from different demulsification treatments were also investigated. Decantation had a larger effect than grinding in terms of phospholipids loss due to alkaline-hydrolysis, indicating this processing step was vital for the manipulation of phospholipids levels remained in oil. Over 80% of initial phospholipids were lost during AEP and only 19.8% of initial phospholipids ended up in cream, skim and sediment phase. 52.55% of the remained phospholipids trapped in cream phase. Just 22.16-32.61 mg/kg phosphorus content could be detected in crude oils, which indicated the separation of phospholipids from the cream phase into aqueous medium. Degumming was not essential in AEP of peanut and the waste generated after demulsification could be a source of phospholipids.

Dissection of the genetic basis of oil content in Chinese peanut cultivars through association mapping.

BACKGROUND: Peanut is one of the primary sources for vegetable oil worldwide, and enhancing oil content is the main objective in several peanut breeding programs of the world. Tightly linked markers are required for faster development of high oil content peanut varieties through genomics-assisted breeding (GAB), and association mapping is one of the promising approaches for discovery of such associated markers. RESULTS: An association mapping panel consisting of 292 peanut varieties extensively distributed in China was phenotyped for oil content and genotyped with 583 polymorphic SSR markers. These markers amplified 3663 alleles with an average of 6.28 alleles per locus. The structure, phylogenetic relationship, and principal component analysis (PCA) indicated two subgroups majorly differentiating based on geographic regions. Genome-wide association analysis identified 12 associated markers including one (AGGS1014_2) highly stable association controlling up to 9.94% phenotypic variance explained (PVE) across multiple environments. Interestingly, the frequency of the favorable alleles for 12 associated markers showed a geographic difference. Two associated markers (AGGS1014_2 and AHGS0798) with 6.90-9.94% PVE were verified to enhance oil content in an independent RIL population and also indicated selection during the breeding program. CONCLUSION: This study provided insights into the genetic basis of oil content in peanut and verified highly associated two SSR markers to facilitate marker-assisted selection for developing high-oil content breeding peanut varieties.

Combination of Alcalase 2.4 L and CaCl2 for aqueous extraction of peanut oil.

The combined application of CaCl2 and Alcalase 2.4 L to the aqueous extraction process of peanuts was evaluated as a method to destabilize the oil body (OB) emulsion and improve the oil yield. After adding 5 mM CaCl2 , the oil yield was reached to 92.0% which was similar with that obtained using Alcalase 2.4 L alone, and the required enzyme loading was decreased by approximately 60 times. In addition, the demulsification mechanism during aqueous extraction process was also investigated. Particle size and zeta-potential measurements indicated that the stability of the peanut OB emulsion dramatically decreased when CaCl2 was added. Under these conditions, the demulsification of Alcalase 2.4 L performed was more efficiently. SDS-PAGE results showed that adding CaCl2 changed the subunit structure of the peanut OB interface proteins and promoted the cross-linking among the arachin Ara h3 isoforms, resulting in unstable emulsions.

Steady expression of high oleic acid in peanut bred by marker-assisted backcrossing for fatty acid desaturase mutant alleles and its effect on seed germination along with other seedling traits.

Peanut (Arachis hypogaea L.) is an important nutrient-rich food legume and valued for its good quality cooking oil. The fatty acid content is the major determinant of the quality of the edible oil. The oils containing higher monounsaturated fatty acid are preferred for improved shelf life and potential health benefits. Therefore, a high oleic/linoleic fatty acid ratio is the target trait in an advanced breeding program. The two mutant alleles, ahFAD2A (on linkage group a09) and ahFAD2B (on linkage group b09) control fatty acid composition for higher oleic/linoleic ratio in peanut. In the present study, marker-assisted backcrossing was employed for the introgression of two FAD2 mutant alleles from SunOleic95R into the chromosome of ICGV06100, a high oil content peanut breeding line. In the marker-assisted backcrossing-introgression lines, a 97% increase in oleic acid, and a 92% reduction in linoleic acid content was observed in comparison to the recurrent parent. Besides, the oleic/linoleic ratio was increased to 25 with respect to the recurrent parent, which was only 1.2. The most significant outcome was the stable expression of oil-content, oleic acid, linoleic acid, and palmitic acid in the marker-assisted backcrossing-introgression lines over the locations. No significant difference was observed between high oleic and normal oleic in peanuts for seedling traits except germination percentage. In addition, marker-assisted backcrossing-introgression lines exhibited higher yield and resistance to foliar fungal diseases, i.e., late leaf spot and rust.

An Automated and High-Throughput Immunoaffinity Magnetic Bead-Based Sample Clean-Up Platform for the Determination of Aflatoxins in Grains and Oils Using UPLC-FLD.

Sample clean-up remains the most time-consuming and error-prone step in the whole analytical procedure for aflatoxins (AFTs) analysis. Herein, an automated and high-throughput sample clean-up platform was developed with a disposable, cost-effective immunoaffinity magnetic bead-based kit. Under optimized conditions, the automated method takes less than 30 min to simultaneously purify 20 samples without requiring any centrifugation or filtering steps. When coupled to ultra-high performance liquid chromatography with fluorescence detection, this new analysis method displays excellent accuracy and precision as well as outstanding efficiency. Furthermore, an interlaboratory study was performed in six laboratories to validate the novel protocol. Mean recovery, repeatability, reproducibility, and Horwitz ratio values were within 91.9%-107.4%, 2.5%-7.4%, 2.7%-10.6%, and 0.26%-0.90, respectively. Results demonstrate that the developed sample clean-up platform is a reliable alternative to most widely adopted clean-up procedures for AFTs in cereals and oils.

Simultaneous determination of carbendazim and chlorothalonil pesticide residues in peanut oil using excitation-emission matrix fluorescence coupled with three-way calibration method.

A novel fluorescence application for simultaneous determination of two common fungicide pesticides (carbendazim and chlorothalonil) in peanut oil is presented. Using the strategy of combining excitation-emission matrix (EEM) fluorescence with three-way calibration methods, the proposed approach successfully achieved quantitative analysis of carbendazim and chlorothalonil pesticide residues in peanut oil, even with highly overlapped spectra. It needs little preparation, using "mathematical separation" instead of "analytical separation" to achieve concentration prediction of target analytes in complex systems. Each analyte was performed using fluorescence spectroscopy after instrument spectral correction and scatter removal. Then the data were modeled with two three-way calibration algorithms, including alternating trilinear decomposition (ATLD) and alternating penalty trilinear decomposition (APTLD). The results indicated that APTLD performed slightly better than ATLD for this system. The carbendazim and chlorothalonil can be recognized simultaneously with the correlation coefficients all above 0.96 between resolved spectra and actual spectra. Satisfactory results have been achieved with the average recoveries (mean ±â€¯standard deviation) of carbendazim and chlorothalonil being 100.2 ±â€¯6.7% and 99.7 ±â€¯6.7%, respectively. Moreover, as for carbendazim and chlorothalonil, the sensitivity (SENs) are 1.50 × 102 and 3.80 × 102 mL ng-1, the limits of detection (LODs) are 11 ng mL-1 and 4.3 ng mL-1, the limit of quantitation (LOQ) are 33.33 ng mL-1 and 13.03 ng mL-1, respectively. The above results demonstrated that the proposed method is sensitive, fast and accurate for direct quantitative analysis of multiple pesticide residues in complex matrix such as that of peanut oil.

Enzyme-free amplified and ultrafast detection of aflatoxin B1 using dual-terminal proximity aptamer probes.

Aptamer probes provide an opportunity for achieving rapid and on-site detection of food contaminants. Herein, we proposed a general design strategy for aptamer probes enabling enzyme-free amplified, ultrafast and one-test tube homogeneous detection of aflatoxin B1 (AFB1). The key feature of the aptamer probe is designed with dual-terminal proximity structures, allowing the binding of one molecule to light up two fluorophores, leading to enzyme-free amplification and a remarkable improvement of signal to background ratio and sensitivity for AFB1 detection. This aptamer probe could accommodate quick response to AFB1, and the detection process could be finished within 1 min, ranking one of the quickest assays for AFB1. AFB1 detection of broad bean paste and peanut oil conferred satisfactory recoveries ranging from 90.3% to 114.8%. Contributed to the generality and simplicity of the design strategy, this structure-switching probe could potentially act as a general platform of on-site detection for food safety.

Electrochemical aptasensor for aflatoxin B1 based on smart host-guest recognition of ß-cyclodextrin polymer.

Developing a simple and reliable method for the detection of the highly concerning mycotoxin, aflatoxin B1 (AFB1), is of great importance to food safety monitoring. In this study, a simple electrochemical aptasensor was presented for the detection of AFB1 based on the host-guest recognition between ferrocene and ß-cyclodextrin (ß-CD). Fc-labeled aptamer of AFB1 first hybridized with its complementary Fc-cDNA. Two ferrocene molecules were brought closely together and couldn't enter into the cavity of ß-CD modified on the electrode. Negligible signal could be observed. Once AFB1 captured the aptamer from the AFB1-sensitive dsDNA, Fc-cDNA was released and subsequently entered into the cavity of ß-CD to form inclusion complexes, giving rise to an distinct increase of Ret and peak current because of the molecular recognition of ß-CD. AC impedance method is more sensitive than DPV method. The electrochemical aptasensor displayed a sensitive response to AFB1 in a wide linear range of 0.1 pg/mL to 10 ng/mL, with a low detection limit of 0.049 pg/mL (0.147 pmol/mL) by AC impedance detection, which is 10-100 lower than previously reported methods. The aptasensor has good selectivity and reliability, which has been successfully applied to the determination of AFB1 in real peanut oil samples with recoveries ranging from 94.5% to 106.7% and inter-assay RSD lower than 11.51%.

Rapid magnetic solid-phase extraction based on alendronate sodium grafted mesoporous magnetic nanoparticle for the determination of trans-resveratrol in peanut oils.

In the present study, a rapid and effective method based on alendronate sodium grafted mesoporous magnetic nanoparticle (Fe3O4@ANDS) extraction for the determination of trans-resveratrol (TRA) in peanut oils was developed by coupling with HPLC-UV detection. The Fe3O4@ANDS was prepared via Lewis acid/base interaction which was simply carried out in mild aqueous condition without the using of organic solvent. The resultant Fe3O4@ANDS encompassed amino group on its surface, and it was employed as magnetic solid-phase extraction adsorbent for purification and enrichment of TRA from peanut oils through hydrogen bond interaction. Under the optimized conditions, the whole pretreatment process could be accomplished within 10 min without time-consuming concentrated and reconstituted process. The linearity range of the proposed method was 1-10,000 ng/g with satisfactory correlation coefficient (R2) of 0.9992. The recoveries in spiked oil samples were in the range of 78.6-118.9% with the RSDs less than 3.3% (intra-day) and 15.2% (inter-day). The limit of detection for TRA in peanut oils was 0.3 ng/g which was comparative to the reported methods by using LC-MS/MS detection. Finally, the established method was successfully applied to the analysis of TRA in several peanut oils with different brands from local market as well as other kinds of vegetable oils.

Dual-Terminal Stemmed Aptamer Beacon for Label-Free Detection of Aflatoxin B1 in Broad Bean Paste and Peanut Oil via Aggregation-Induced Emission.

Aflatoxin B1 (AFB1) contamination ranks as one of the most critical food safety issues, and assays for its on-site monitoring are highly demanded. Herein, we propose a label-free, one-tube, homogeneous, and cheap AFB1 assay based on a finely tunable dual-terminal stemmed aptamer beacon (DS aptamer beacon) and aggregation-induced emission (AIE) effects. The DS aptamer beacon structure could provide terminal protection of the aptamer probe against exonuclease I and confer specific and quick response to target AFB1. In comparison to the conventional molecule beacon structure, the stability of the DS aptamer beacon could be finely tuned by adjusting its two terminal stems, allowing for elaborately optimizing probe affinity and selectivity. By the utilization of an AIE-active fluorophore, which would be lighted up by aggregating to negatively charged DNA, AFB1 could be determined in a label-free manner. The proposed method could quantify AFB1 in one test tube using two unlabeled DNA strands. It has been successfully applied for analyzing AFB1 in peanut oil and broad bean sauce, with total recoveries ranging from 92.75 to 118.70%. Thus, the DS aptamer beacon-based assay could potentially facilitate real-time monitoring and controlling of AFB1 pollution.

A novel method for detection of camellia oil adulteration based on time-resolved emission fluorescence.

In this study, time-resolved emission fluorescence (TRES) combined with chemometrics was developed and employed for adulteration analysis of camellia oil. TRES was first decomposed by parallel factors analysis (PARAFAC). Next, an artificial neural network (ANN) model was built for the adulteration analysis. A linear range of 5-50%, a limit of detection (LOD) of 3% and root mean square error of prediction (RMSEP) values lower than 3% were achieved. Compared with the steady-state measurement, easy access to the information from fluorophores of low concentration was shown to be an intrinsic advantage of the time-resolved measurement; this advantageous characteristic was helpful for optimizing adulteration analysis. It was demonstrated that TRES combined with chemometrics was a simple, rapid and non-intrusive method for adulteration analysis of vegetable oil.

Fatty Acid Profiles of Various Vegetable Oils and the Association between the Use of Palm Oil vs. Peanut Oil and Risk Factors for Non-Communicable Diseases in Yangon Region, Myanmar.

The majority of vegetable oils used in food preparation in Myanmar are imported and sold non-branded. Little is known about their fatty acid (FA) content. We aimed to investigate the FA composition of commonly used vegetable oils in the Yangon region, and the association between the use of palm oil vs. peanut oil and risk factors for non-communicable disease (NCD). A multistage cluster survey was conducted in 2016, and 128 oil samples from 114 households were collected. Data on NCD risk factors were obtained from a household-based survey in the same region, between 2013 and 2014. The oils most commonly sampled were non-branded peanut oil (43%) and non-branded palm oil (19%). Non-branded palm oil had a significantly higher content of saturated fatty acids (36.1 g/100 g) and a lower content of polyunsaturated fatty acids (9.3 g/100 g) than branded palm oil. No significant differences were observed regarding peanut oil. Among men, palm oil users had significantly lower mean fasting plasma glucose levels and mean BMI than peanut oil users. Among women, palm oil users had significantly higher mean diastolic blood pressure, and higher mean levels of total cholesterol and triglycerides, than peanut oil users. Regulation of the marketing of non-branded oils should be encouraged.

Hyphenation of supercritical fluid chromatography with tandem mass spectrometry for fast determination of four aflatoxins in edible oil.

RATIONALE: Aflatoxins (AFTs) are of great concern all over the world. Supercritical fluid chromatography (SFC) has the advantage of fast, high resolution and excellent compatibility with a broad range of organic solvents and samples, thus hyphenating SFC with tandem mass spectrometry (MS/MS) can be used for the easy and fast determination of AFTs in edible oils. METHODS: Edible oil was spiked with isotope-labeled aflatoxin standards, diluted with hexane and extracted with acetonitrile. The extraction was directly loaded to an SFC apparatus and separated on a UPC(2) 2-EP column with CO2 -methanol gradient elution. A post-column make-up flow was introduced to facilitate mass spectrometry performance, and the mixture was analyzed by MS/MS with an electrospray ionization (ESI) source. RESULTS: The SFC conditions including separation column, modifier and sample solvent were optimized, and the four target aflatoxins were baseline separated. The ESI interface parameters were also investigated, implicating the make-up flow as a critical factor for sensitive determination by SFC-MS/MS. The LOQs for the AFTs were 0.05-0.12 µg L(-1) , while the RSDs were lower than 8.5%. CONCLUSIONS: Supercritical fluid chromatography was successfully coupled to tandem mass spectrometry to establish a simple, fast and sensitive method for the analysis of four aflatoxins in edible oil. This shows the combination of SFC-MS/MS has great potential in determination of trace contaminants in food. Copyright © 2016 John Wiley & Sons, Ltd.