The double flower variant of yellowhorn is due to a LINE1 transposon-mediated insertion.

As essential organs of reproduction in angiosperms, flowers, and the genetic mechanisms of their development have been well characterized in many plant species but not in the woody tree yellowhorn (Xanthoceras sorbifolium). Here, we focused on the double flower phenotype in yellowhorn, which has high ornamental value. We found a candidate C-class gene, AGAMOUS1 (XsAG1), through bovine serum albumin sequencing and genetics analysis with a Long Interpersed Nuclear Elements 1 (LINE1) transposable element fragment (Xsag1-LINE1-1) inserted into its second intron that caused a loss-of-C-function and therefore the double flower phenotype. In situ hybridization of XsAG1 and analysis of the expression levels of other ABC genes were used to identify differences between single- and double-flower development processes. These findings enrich our understanding of double flower formation in yellowhorn and provide evidence that transposon insertions into genes can reshape plant traits in forest trees.

A computational and experimental study of cis-trans isomeric pesticides based on collision-induced dissociation of high-resolution mass spectrometry.

RATIONALE: Pesticide isomers are widely available in agricultural production and may vary widely in biological activity, potency, and toxicity. Chromatographic and mass spectrometric analysis of pesticide isomers is challenging due to structural similarities. METHODS: Based on liquid chromatography time-of-flight mass spectrometry, identification of cis-trans isomeric pesticides was achieved through retention time, characteristic fragment ions, and relative abundance ratio. Furthermore, theoretical and basic research has been conducted on the differences in characteristic fragment ions and their relative abundance ratios of cis-trans isomers. On the one hand, the cleavage pathways of six cis-trans isomers were elucidated through collision-induced dissociation to explain different fragment ions of the isomers. On the other hand, for those with the same fragment ions but different abundance ratios, energy-resolved mass spectrometry combined with computational chemical density functional theory in terms of kinetics, thermodynamics, and bond lengths was employed to explain the reasons for the differences in characteristic fragment ions and their abundance ratios. RESULTS: A high-resolution mass spectrometry method was developed for the separation and analysis of cis-trans isomers of pesticides in traditional Chinese medicine Radix Codonopsis, and six pesticide isomers were distinguished by retention time, product ions, and relative abundance ratios. The limits of quantification of the six pesticides were up to 10 µg/kg, and the linear ranges of them were 10-200 µg/kg, with coefficients of determination (R2) > 0.99, which demonstrated the good linearity of the six pesticides. The recoveries of the pesticides at spiked concentrations of 10, 20, and 100 µg/kg reached 70-120% with relative standard deviations ≤20%. CONCLUSIONS: It was demonstrated that the application of the method was well suited for accurate qualitative and quantitative analysis for isomers with different structures, which could avoid false-negative results caused by ignoring other isomers effectively.

One-Pot Controllable Assembly of a Baicalin-Condensed Aptamer Nanodrug for Synergistic Anti-Obesity.

The rapid, simple and low-cost preparation of DNA micro-nano-architectures remain challenging in biosensing and therapy. Polymerase chain reaction (PCR)-driven DNA micro-nano-flowers are used to construct a nanosized baicalin-compressed-aptamer-nanodrug (bcaND) via one-pot assembly for targeted and synergistic anti-obesity. In the design, the tailored Adipo-8 (tAdi-8) overhang in the PCR amplicon displays anti-obesity targeting activity, while the baicalin loaded in the bcaND by embedding the amplicon plays a three-fold role as a lipid-lowering factor, bcaND size compressor, and uncoupling protein-1 (UCP1)-raised thermogenic activator. The ingenious bcaND represents an advanced multifunctional nanomaterial capable of adjusting the morphology at an optimal 400/1 molar ratio of Mg2+ to phosphate groups, compressing the size from 2.699 µm to 214.76 nm using 1 mg/mL baicalin at a temperature of 70 °C, an effective payload with amplicons of up to 98.94%, and a maximum baicalin load of 86.21 g/g DNA. Responsive release in acidic conditions (pH 5.0) occurs within 72 h, accelerating thermogenesis via UCP1 up-regulation by 2.5-fold in 3T3-L1-preadipocytes and 13.7-fold in the white-adipose-tissue (WAT) of mice, targeting adipocytes and visceral white adipose tissue. It plays an efficient synergistic role in obesity therapy in vitro and in vivo, providing a new direction for DNA self-assembly nanotechnology.

Advanced screening and tailoring strategies of pesticide aptamer for constructing biosensor.

The rapid development of aptamers has helped address the challenges presented by the wide existed pesticides contaminations. Screening of aptamers with excellent performance is a prerequisite for successfully constructing biosensors, while further tailoring of aptamers with enhanced activity greatly improved the assay performance. Firstly, this paper reviewed the advanced screening strategies for pesticides aptamers, including immobilization screening that preserves the native structures of targets, non-immobilized screening based on nanomaterials, capillary electrophoresis-systematic evolution of ligands by exponential enrichment (CE-SELEX), virtual screening in silico, high-throughput selection, and rational secondary library generation methods, which contributed significantly to improve the success rate of screening, reduce the screening time, and ensure aptamer binding affinity. Secondly, the precise tailoring strategies for pesticides aptamers were modularly elaborated, containing deletion, splitting, elongation, and fusion, which provided various advantages like cost-efficiency, enhanced binding affinity, and new derived functional motifs. Thirdly, the developed aptamer-based biosensors (aptasensors) for pesticide detection were systematically reviewed according to the different signal output modes. Finally, the challenges and future perspectives of pesticide detection are discussed comprehensively.

Residual levels and dietary exposure risk assessment of banned pesticides in fruits and vegetables from Chinese market based on long-term nontargeted screening by HPLC-Q-TOF/MS.

The negative impact of banned pesticides is of special importance for their high toxicity. In this study, nationwide screening of banned pesticides in 37462 fruit and vegetable samples was carried out from 2012 to 2018 using a self-developed HPLC-Q-TOF/MS technique. The dietary exposure risks associated with the banned pesticides were assessed. The results showed that 66.62 % of the samples were detected at least one pesticide. Among the pesticide-positive samples, a total of 18 banned pesticides were detected in 1798 samples for 1896 times. The risk assessment revealed that 11.71 % of the positive detections exceeded the safety limits and posed an unacceptable risk, while 37.29 % of the positive detections posed acceptable risks. According to the screening and assessment results, two national maps were presented to show the total detection ratios of the banned pesticides and the unacceptable risks of dietary exposure. It should be noted that omethoate had higher residual concentration, unacceptable risk frequency and unacceptable risk proportion. This is the first nationwide comprehensive report on screening and risk assessment banned pesticides.

Risk assessment of carbofuran residues in fruits and vegetables at the Chinese market: A 7-year survey.

The United Nations designated 2021 as the International Year of Fruits and Vegetables (IYFV), with the goal of educating populations regarding the role of such produce in nutrition, food safety, and overall health. Carbofuran is a highly toxic insecticide and nematocide, and its use to treat fruit trees, vegetables, tea, and medicinal herbs is thus prohibited. However, carbofuran residues are still detectable via LC-Q-TOF/MS in fruit and vegetable samples collected from 138 sites in 31 regions. In the present study, carbofuran levels were sampled at 1388 sampling sites in 31 regions (provinces, autonomous regions, and municipalities) not including Hong Kong, Macao, or Taiwan. In total, over 36,000 samples (including 12,547 samples of 41 kinds of fruits and 23,785 samples of 83 kinds of vegetables) were randomly collected from supermarkets and farmer's markets. These data were used to conduct a risk assessment pertaining to dietary carbofuran exposure through the consumption of fruits and vegetables. In total, carbofuran residues were detectable in 2.0% of fruits and 2.3% of vegetables. Risk assessments indicated that the intake of fruits and vegetables harboring carbofuran residues did not pose a chronic health risk. However, peaches, grapes, sweet peppers, celery, Chinese chives, leaf lettuce, spinach, small rape, mustard greens, cucumbers, watermelons, Chinese wolfberry leaves, wax gourds, snap beans, bitter melons, green Chinese vegetables, lettuce, shallot, cowpeas, eggplants, tomatoes, tangerines, summer squash, oranges, lemons, Chinese cabbage, peppers, and strawberries were associated with an unacceptable acute risk to both children and adults. Moreover, crown daisies, nectarines, citrus fruits, pitayas, melons, kale, cabbages, milk Chinese cabbage, carrots, and melons were associated with an unacceptable acute risk to children. Substantial acute risk to children and adults was observed for fruits and vegetables from surveyed regions other than Inner Mongolia, Yunnan, Liaoning, Fujian, Xinjiang, and Hubei. Together, these data provide a foundation for future research aimed at the management of carbofuran residues in fruits and vegetables in an effort to better protect consumer health.

Simultaneous determination and risk assessment of highly toxic pesticides in the market-sold vegetables and fruits in China: A 4-year investigational study.

This study investigated the levels of highly toxic pesticides (HTPs) in 6554 vegetable and fruit samples from 31 regions of China, along with the associated risk of dietary exposure for the population between 2014 and 2017. 18 HTPs were detected in 325 (4.96%) samples, and the levels of HTPs in 103 (1.57%) samples were found to be higher than the maximum residue limits (MRLs) of China. The rate of detection of HTPs in six types of vegetables and fruits, in a decreasing order, was found to be as follows: eggplant (8.84%) >grape (5.58%) >tomato (5.43%) >cucumber (5.43%) >pear (3.12%) >apple (2.30%). The level of contamination of HTPs was found to be higher in vegetables compared with fruits. The vegetable and fruit samples with the highest percentages of HTPs exceeding MRLs were found in eggplants from Guangxi (20%) and grapes from Inner Mongolia (12.5%), respectively. Both, the average target hazard quotient (THQ) of a single highly toxic pesticide (HTP) and the average hazard index (HI) of the mixture of HTPs for adults and children from vegetables and fruits from the 31 regions were found to be less than one. Omethoate, carbofuran, ethoprophos, triazophos, and phorate were identified as the major contributors to the average HI for vegetables, and carbofuran, ethoprophos, omethoate, phorate, and phosphamidon were identified as the primary contributors to the average HI for fruits. The results of this study revealed that HTPs in vegetables and fruits did not cause any significant chronic risk of dietary exposure. The detection of HTPs exceeding MRLs in some of the samples implied that appropriate management guidelines for HTPs should be implemented to protect the health of the consumers.

Discrimination of botanical origins for Chinese honey according to free amino acids content by high-performance liquid chromatography with fluorescence detection with chemometric approaches.

BACKGROUND: The contents of 18 free amino acids in 87 Chinese honey samples from four botanical origins (linden, acacia, vitex and rape) were determined by developing a high-performance liquid chromatography with fluorescence detector (HPLC-FLD) method with an in-loop automated pre-column derivatization. The free amino acid profiles of these samples were used to construct a statistical model to distinguish honeys from various floral origins. RESULTS: The average contents of all free amino acids in linden honey were lower than in the other three types of honey. Phenylalanine was particularly useful in the present study because its average content in vitex honey was far higher than in any other honey samples. There is no doubt that both phenylalanine and tyrosine can be considered as the marker free amino acid in Chinese vitex honey. Principal component analysis (PCA) was conducted based on 15 free amino acids and showed significant differences among the honey samples. The cumulative variance for the first two components was 80.62%, and the four principal components can explain 94.18% of the total variance. In the two first component scores, the honey samples can be separated according to their botanical origins. Cluster analysis of amino acid data also revealed that the botanical origins of honey samples correlated with their amino acid content. Back-propagation artificial neural network (BP-ANN) and naïve Bayes methods were employed to construct the classification models. The results revealed an excellent separation among honey samples according to their botanical origin with 100% accuracy in model training for both BP-ANN and naïve Bayes. CONCLUSION: It indicated that the free amino acid profile determined by HPLC-FLD can provide sufficient information to discriminate honey samples according to their botanical origins. © 2016 Society of Chemical Industry.

Wide-scope screening of pesticides in fruits and vegetables using information-dependent acquisition employing UHPLC-QTOF-MS and automated MS/MS library searching.

This paper presents an application of ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) for simultaneous screening and identification of 427 pesticides in fresh fruit and vegetable samples. Both full MS scan mode for quantification, and an artificial-intelligence-based product ion scan mode information-dependent acquisition (IDA) providing automatic MS to MS/MS switching of product ion spectra for identification, were conducted by one injection. A home-in collision-induced-dissociation all product ions accurate mass spectra library containing more than 1700 spectra was developed prior to actual application. Both qualitative and quantitative validations of the method were carried out. The result showed that 97.4 % of the pesticides had the screening detection limit (SDL) less than 50 µg kg-1 and more than 86.7 % could be confirmed by accurate MS/MS spectra embodied in the home-made library. Meanwhile, calibration curves covering two orders of magnitude were performed, and they were linear over the concentration range studied for the selected matrices (from 5 to 500 µg kg-1 for most of the pesticides). Recoveries between 80 and 110 % in four matrices (apple, orange, tomato, and spinach) at two spiked levels, 10 and 100 µg kg-1, was 88.7 or 86.8 %. Furthermore, the overall relative standard deviation (RSD, n = 12) for 94.3 % of the pesticides in 10 µg kg-1 and 98.1 % of the pesticides in 100 µg kg-1 spiked levels was less than 20 %. In order to validate the suitability for routine analysis, the method was applied to 448 fruit and vegetable samples purchased in different local markets. The results show 83.3 % of the analyzed samples have positive findings (higher than the limits of identification and quantification), and 412 commodity-pesticide combinations are identified in our scope. The approach proved to be a cost-effective, time-saving and powerful strategy for routine large-scope screening of pesticides.

High-Throughput Analytical Techniques for Determination of Residues of 653 Multiclass Pesticides and Chemical Pollutants in Tea--Part V: A Comparative Study of the Influence of Tea Hydration on the Efficiency of Pesticide Multiresidue Determination Using Three Sample Preparation Methods and GC/MS/MS.

This paper describes a comparative study of the influence of three sample preparation techniques (M1: hydration+oscillating extraction+partial extraction solution hexane partitioning cleanup; M2: hydration+oscillating extraction+overall extraction solution SPE cleanup; and M3: pure acetonitrile homogeneous extraction+overall extraction SPE cleanup) on the determination efficiency of 456 pesticide multiresidues in tea. First, it was discovered from the mathematical correlation equation of 329 pesticide recoveries established and log Kow values that the extraction efficiency of hydration method M1 has obvious correlation with pesticide log Kow, making the extraction efficiency of M1 take the shape of an arc trend line with a certain arc hanging down from both ends of polar pesticides and nonpolar pesticides. Second, regarding the M1 method, the interfering matter after co-extraction increased in large quantities, which markedly lowered the S/N of the target pesticides and method sensitivity, leading to an obvious decrease of the method efficiency. The fortification experiment of the uniform limit 0.010 mg/kg proved that with the M1 hydration method there are 23 pesticides with recoveries between 70 and 120% and RSD<20%, accounting for only 5.0%, while with nonhydration method M3 there are 229 pesticides, making up 50%.

High Throughput Analytical Techniques for the Determination and Confirmation of Residues of 653 Multiclass Pesticides and Chemical Pollutants in Tea by GC/MS, GC/MS/MS, and LC/MS/MS: Collaborative Study, First Action 2014.09.

Thirty laboratories from fom North and South America, Europe, and Asia participated in this AOAC collaborative study (15 from China; five from Germany; two each from Italy and the United States; and one each from the Republic of Korea, Canada, Spain, Japan, Belgium, and India). Participants represented government regulatory, commercial testing, university, research institute, and private laboratories. The single-laboratory validated (SLV) tea method was evaluated in the collaborative study to determine the recovery and reproducibility of the method under multilaboratory conditions. Since there were no restrictions regarding the type of analytical instrumentation to use for the analyses, laboratories used a combination of equipment that included GC/MS, GC/MS/MS, and LC/MS/MS instruments from 22 different manufacturers, 21 brands of GC and LC columns, 13 different GC temperature programming profiles, 11 LC gradient elution programs, and six different vendor manufactured SPE cartridges. Even though all the analytical performance parameters for all the 653 compounds had been determined in the SLV study, guidance was obtained from an expert review panel of the AOAC Method-Centric Committee on Pesticide Residues to conduct the multilaboratory collaborative study based on 20 selected compounds that can be analyzed by GC/MS and 20 compounds that can be analyzed by LC/MS/MS. Altogether, 560 samples covering the 40 selected pesticides were analyzed in the study. These samples included green tea and oolong tea samples fortified typically at the European Union maximum residue limit for regulatory guidance and compliance, aged tea samples incurred with 20 pesticides, and green tea and oolong tea samples incurred with five pesticides. The analysis of the 560 samples generated a total of 82 459 test results by the 30 participating laboratories. One laboratory failed to meet the proficiency requirements in the precollaborative study. Therefore, its data submitted for the collaborative study were excluded from further analysis and interpretation. The results presented are therefore the 6638 analytical results obtained from the 29 remaining laboratories, which included 1977 results generated by GC/MS, 1704 results by GC/MS/MS, and 2957 results by LC/MS/MS. It was determined after application of the Grubbs and Dixon tests for outliers to the data sets that there were 65 outlier results from the 1977 GC/MS results (3.3%), 65 outlier results from the 1704 GC/MS/MS results (3.8%), and 57 outlier results out of 2957 LC/MS/MS results (1.9%), representing 0.98, 0.98, and 0.86%, respectively, of the 6638 results generated in the study. Analysis with the AOAC statistical software package also confirmed that the method is rugged, and average recovery, average concentration, RSDr, RSDR, and HorRat values all meet recovery and reproducibility criteria for use in multiple laboratories. The Study Director is recommending this method for adoption as an AOAC First Action Official MethodSM.

High-Throughput Analytical Techniques for Multiresidue, Multiclass Determination of 653 Pesticides and Chemical Pollutants in Tea-Part IV: Evaluation of the Ruggedness of the Method, Error Analysis, and Key Control Points of the Method.

A 3 month study was conducted on the ruggedness of a multiresidue method for accuracy and stability. The results indicate that in terms of Youden pair ratios of 201 pesticide aged tea samples falling approximately within 1.00-1.20 of the ratio of theoretical spraying concentrations, the differences do not exceed 5% for percentages made up by ratios of the fixed values obtained by two kinds of instruments for two teas and those made up by 18 circular determinations. However, regarding two kinds of SPE cartridges, the Cleanert TPT cartridge is higher than the ENVI-CARB+primary secondary amine (PSA) cartridge by 10%. Pertaining to RSD values of "parallel samples" and whether it is green tea or Woolong tea, the percentages of RSD≤15% values of the parallel samples all exceed 88%. Whether it is the first or circular determination for two teas and analytical results from two kinds of instruments, the percentages of RSD≤15% values have a difference of less than 6%, while the TPT cartridge is better than ENVI-CARB+PSA by above 6% for the two cartridges. Concerning RSDs of Youden pair ratios, RSD≤15% values have a proportion exceeding 85% for both green tea and Woolong tea, and the percentage is greater than 87% whether it is for two kinds of SPE cartridges or two kinds of instruments. In terms of Youden pair ratios and the classified statistical analysis of the ruggedness data of parallel samples, the proportion of RSD≤15% values of Youden pair ratios is 8% higher for the TPT cartridge than the ENVI-CARB+PSA cartridge; the proportion of RSD≤15% values of parallel samples is 6.2% higher for the TPT cartridge than the ENVI-CARB+PSA cartridge. Data show no marked differences for two teas and two kinds of instruments. A comparison of the aforementioned aspects finds that good ruggedness was obtained with both SPE cleanup methods, and the results from the TPT cartridge are better than those from the ENVI-CARB+PSA cartridge.

[Application of ICP-MS method in the determination of mineral elements in vitex honey for the classification of their geographical origins with chemometric approach].

In the present work, the contents of 38 elements of 65 vitex (Vitex negundo var. heterophylla Rehd. ) honey samples from Shunyi of Beijing, Fuping and Pingshan of Hebei province were determined by inductively coupled plasma mass spectrometry (ICP-MS). Among them, B, Na, Mg, P, K, Ca, Fe and Zn were the most abundant elements with mean contents more than 1 mg kg-1. It can be found that there were relationships between the contents of elements and the geographical origin of vitex honey samples. Taking the contents of 29 out of 38 mineral elements (Na, Mg, Al, K, Ti, V, Mn, Fe, Ni, Cu, Zn, Ga, As, Sr, Y, Mo, Cd, Ba, La, Ce, Pr, Nd, Sm, Gd, Dy, Ho, T1, Pb and U) as variables, the chemometric methods, such as principal component analysis (PCA) and back-propagation artificial neural network (BP-ANN), were applied to classify vitex honey samples according to their geographical origins. PCA reduced all of the variables to four principal components and could explain 81. 6% of the total variances. The results indicated that PCA could mainly classify the vitex honey samples into three groups. BP-ANN was explored to construct classification model of vitex honeys according to their geographical origin. For the whole data set, the overall correct classification rate and cross-validation (leave one out method) rate of proposed BP-ANN model was 100% and 95. 4%, respectively. To further test the stability of the model developed for prediction, 75% of honey samples of each geographical origin were randomly selected for the model training set, and the remaining samples were classified with the use of the constructed model. Both the overall correct classification rate and prediction rate of proposed BP-ANN model were 100%. It is concluded that the profiles of multi-element by ICP-MS with chemometric methods could be a potential and powerful tool for the classification of vitex honey samples from different geographical origins.

Determination of trace elements in rape honey and its corresponding rape flower and stem by ICP-MS.

The determination of 10 trace elements including Na, Mg, P, K, Ca, Mn, Zn, Rb, Sr and Ba, in rape honey and its corresponding rape flower and stem gathered from nine sampling sites was carried out by inductively coupled plasma mass spectrometry (ICP-MS). The contents of K, P, Ca, Mg and Na were obviously higher than Zn, Rb, Mn, Sr and Ba in rape honey, rape flower and rape stem. For the first five elements, K had the highest content, followed by P, Ca, Mg and Na. However, the order of content for latter five elements was not the same in different matrixes. The contents of K, P and Ca were all higher than 1 000 mg x kg(-1) in rape flower and rape stem, while the contents of P, Ca, Mn, Zn and Rb in rape flower were slightly higher than in rape stem. It can be concluded that rape flower showed slightly higher concentrating ability for trace elements than rape stem. Based on these results, radar chart was firstly applied to research the relationship of 10 elements in rape honey and its corresponding rape flower and stem. The aim of the present work was to study the possibility of using trace elements contents in rape flower to trace the geographical and botanical origin of honey instead of rape honey. It can be found from the radar charts that the stars of rape honey, rape flower and rape stem were similar to each other. This research not only provides the basic data of trace elements in comparative study of rape honey, but also gives scientific basis for tracing the origin of rape honey according to the trace elements in corresponding rape flower that replaces those of rape honey.

Specific determination of (-)- and (+)-gossypol in vegetable oil by ultra-performance liquid chromatography-tandem mass spectrometry with chiral pre-column derivatization.

BACKGROUND: As the unique ingredient of cottonseed oil, gossypol is toxic and there are differences between the enantiomers. Although the determination of (-)- and (+)-gossypol in cotton plants or cottonseed was performed, the detection about the gossypol enantiomers in vegetable oil was rarely reported. OBJECTIVE: To develop a specific inspection method for (-)- and (+)-gossypol in vegetable oil by ultra-performance liquid chromatography (UPLC) -tandem mass spectrometry with chiral pre-column derivatization. METHOD: (-)- and (+)-gossypol were separated on an ACQUITY C18 reverse phase column (100 mm × 2.1 mm, 1.7 µm) maintained at 40 °C on an UPLC system with gradient elution. The mobile phases was composed of methanol (solvent A) and water containing 0.1% formic acid (solvent B). Analytes were detected with Electron Spray Ionization source in the positive mode. The contents of (-)- and (+)-gossypol standard derivatives were quantified by Multiple Reaction Monitoring with the transitions of m/z 633/483 for quantitative ion pairs and m/z 633/558 for qualitative ion pairs. RESULTS: The results indicated that the quantitative analysis could be accomplished within 12.5 minutes and the limits of detection of (-)- and (+)-gossypol were 25.55 µg/kg and 14.67 µg/kg, respectively. When this method was applied to olive oil spiked with gossypol standards, good recoveries (95.37%-105.84% for (-)-gossypol, 98.96%-105.14% for (+)-gossypol) and RSDs (3.41%-6.02% for (-)-gossypol, 4.50%-6.94% for (+)-gossypol) were achieved. CONCLUSION: The present study confirms the feasibility of determining the gossypol enantiomers and achieves trace determination of (-)- and (+)-gossypol in vegetable oil. These results make this method more suitable for the qualitative identification of whether the vegetable oil has been mixed with cottonseed oil. HIGHLIGHTS: The trace determination of (-)- and (+)-gossypol in vegetable oils has been achieved. This method provides technical support for further identification of whether commercially available vegetable oil has been mixed with cottonseed oil.

Decoding wheat contamination through self-assembled whole-cell biosensor combined with linear and non-linear machine learning algorithms.

The contamination of mycotoxins is a serious problem around the world. It has detrimental effects on human beings and leads to tremendous economic loss. It is essential to develop a rapid and non-destructive method for contamination recognition particularly for early alarm. In this study, the whole-cell biosensor array was constructed and employed for rapid recognition of wheat contamination by combining with machine learning algorithms. Seven key VOCs were explored through univariate coupling to multivariate analysis of orthogonal partial least squares-discrimination analysis (OPLS-DA) models. The promoters of dnaK, katG, oxyR, soxS obtained from the stress-responsive of key VOCs were fused to the bacterial operon and fabricated on the whole-cell biosensor. The constructed whole-cell biosensor array was consisted with four kinds of sensors and 18 sensor unit. The bioluminescent intensity combined with linear machine learning algorithm of partial least squares discriminant analysis (PLS-DA) and non-linear algorithms of back propagating artificial neural network (BP-ANN) and least square support vector machine (LS-SVM) were employed to establish discrimination models for mold contamination especially for early warning. The Monte-Carlo strategy was performed to generate thirty subsets for modeling to give more reliable results. As a result, the whole-cell biosensor combined with non-linear algorithm of LS-SVM was practicable for detecting mold identification for wheat early-warning with the accuracy of 97.24%. Additionally, this study provides practical and effective methods not only for wheat quality guarantee and supervision but also for other foodstuffs.

Rapid determination of volatile benzene derivatives and chlorobenzenes in goat's milk by HS-SPME-GC-MS/MS.

A method for the determination of eight benzenes (BTEXs) and twelve chlorobenzenes (CBs) in goat's milk by headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS/MS) was developed. The study investigated the impact of various factors such as extraction fiber type, salt amount, equilibrium conditions, and desorption conditions on the outcomes. Target analytes were separated on a DB-HeavyWAX column and quantified using the external standard method. The results showed that the target compounds had a good linear relationship in the range of 0.01 ∼ 50 µg/L (R2 > 0.997), the limit of detection (LOD) was 0.003 ∼ 0.150 µg/L, and the limit of quantification (LOQ) was 0.01 ∼ 0.50 µg/L. The average recoveries were 82%-116% and the relative standard deviation (RSD) was 0.8%-17.3% under the three addition levels of 1×, 2×, and 10 × LOQ. In a survey of twenty goat's milk samples, only ethylbenzene, xylenes, cumene, chlorobenzene, and 1,4-dichlorobenzene were detected at levels exceeding their respective limits of quantification. The method was evaluated using two ecological scales (Eco-Scale), GAPI and AGREEN, to verify its environmental friendliness and applicability. This method is simple, green, and efficient, which provides a certain theoretical basis for the production and quality safety evaluation of dairy products.

Simultaneous determination of pesticides, mycotoxins and ferulic acid in Angelica sinensis by GC/LC-Q-TOF/MS.

In this study, a strong applicable method that could determine a total of 33 pesticides (54 compounds), 11 mycotoxins and functional components (ferulic acid) simultaneously in Angelica sinensis was developed. The compatibility of the sample pretreatment method for pesticides, mycotoxins, and functional components was improved by optimizing the acidity of extraction solvents, the sequence of ice bath and oscillation, the volumetric solution, and so on. The PRiME HLB SPE pretreatment method was chosen as the optimal one when comparing four pretreatment methods. Among the 65 contaminants, 38 of those determined by liquid chromatography and 41 of those by gas chromatography, which showed good linearity (R2 > 0.9801), 97 % of them had a limit of quantitation (LOQ) of lower than 0.02 mg kg-1. The recovery of all compounds were suited between 70 % to 120 % and the RSD were all lower than 20 % at the spiked levels of LOQ, 2 × LOQ, and 10 × LOQ. For ferulic acid, the LOQ was 50 ng/mL, and it showed good linearity (R2=0.9988) within the range of 0.5 to 10 µg/mL. The recovery and RSD were 98.1 %, and 3.2 % (n = 6), respectively. The simultaneous determination of cross-category compounds in a single sample preparation was obtained by the combination of SPE and GC/LC-Q-TOF/MS. Therefore, this study could not only shorten the time for data acquisition and data analysis, but also improve the experimental efficiency.

[Rapid determination of 15 N-nitrosamines in air-dried yak meat using one-step QuEChERS-gas chromatography-triple quadrupole mass spectrometry].

A method based on gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS) coupled with one-step QuEChERS technique was developed for the simultaneous determination of 15 N-nitrosamines in air-dried yak meat. The hydration volume, extraction solvent, extracting salt, and cleaning material were optimized according to the characteristics of the N-nitrosamines and sample matrix. The optimized conditions were as follows: 10 mL of purified water for sample hydration, acetonitrile as the extraction solvent for the sample after hydration, 4.0 g of anhydrous MgSO4 and 1.0 g of NaCl as extracting salts, 500 mg of MgSO4+25 mg of C18+50 mg of PSA as cleaning materials. Favorable recoveries of the 15 N-nitrosamines were obtained when the extraction solution was incompletely dried. Thus, the final extract was dried to below 0.5 mL under a mild nitrogen stream and then redissolved to 0.5 mL with acetonitrile. After filtration, 200 µL of the sample was transferred to an autosampler vial for GC-MS/MS analysis. The 15 N-nitrosamines were determined using GC-MS/MS on a DB-HeavyWAX column (30 m×0.25 mm×0.25 µm) with an electron impact ion source in multiple-reaction monitoring (MRM) mode, and quantified using an external standard method. Under the optimized experimental conditions, the results showed that the calibration curves exhibited good linearities for the 15 N-nitrosamines, with correlation coefficients (r2) greater than 0.9990. The limits of detection (LODs) and the limits of quantification (LOQs) ranged from 0.05 to 0.20 µg/kg and from 0.10 to 0.50 µg/kg, respectively. At spiked levels of 1LOQ, 2LOQ, and 10LOQ, the average recoveries were 79.4%-102.1%, 80.6%-109.5%, and 83.0%-110.6%, respectively, and the relative standard deviations were in the range of 0.8%-16.0%. The low matrix effects of the 15 N-nitrosamines indicated the high sensitivity of the proposed method. The method was applied to detect representative commercial air-dried yak meat samples obtained using different processing techniques. Seven N-nitrosamines, including N-nitrosodimethylamine, N-nitrosodiisobutylamine, N-nitrosodibutylamine, N-methyl-N-phenylnitrous amide, N-ethyl-N-nitrosoaniline, N-nitrosopyrrolidine, and N-nitrosodiphenylamine were detected in all samples. The average contents of the seven N-nitrosamines was 0.08-20.18 µg/kg. The detection rates and average contents of the N-nitrosamines in cooked air-dried yak meat samples were higher than those in traditional raw air-dried yak meat samples. Compared with the manual QuEChERS method, the one-step QuEChERS method developed integrated the extraction and clean-up procedures into one single run, and the detection efficiency was considerably improved. The developed method is simple, rapid, highly sensitive, and insusceptible to human errors. Thus, it is useful for the determination of N-nitrosamines in air-dried yak meat and can be extended to the qualitative and quantitative analysis of N-nitrosamines in other meat products. It also provides method support and a data reference for the general determination of N-nitrosamines, which is of great significance for food safety.