Development of Pure Certified Reference Material of Cannabidiol.

Cannabidiol (CBD) is the major functional component in hemp and has a broad range of pharmacological applications, such as analgesic, anti-epileptic, anti-anxiety, etc. Currently, CBD is widely used in pharmaceuticals, cosmetics, and food. To ensure the quality and safety of the products containing CBD, more and more related sample testing is being conducted, and the demand for CBD-certified reference material (CRM) has also sharply increased. However, there is currently a lack of relevant reference materials. In this paper, a simple method for preparing CBD CRM was established based on preparative liquid chromatography using crude hemp extract as a raw material. A qualitative analysis of CBD was performed using techniques such as ultraviolet absorption spectroscopy (UV), infrared spectroscopy (IR), mass spectrometry (MS), nuclear magnetic resonance spectroscopy (NMR), and differential scanning calorimetry (DSC). High-performance liquid chromatography (HPLC) was used for the homogeneity and stability tests, and the data were analyzed using an F-test and a T-test, respectively. Then, eight qualified laboratories were chosen for the determination of a certified value using HPLC. The results show that the CBD CRM had excellent homogeneity and good stability for 18 months. The certified value was 99.57%, with an expanded uncertainty of 0.24% (p = 0.95, k = 2). The developed CBD CRM can be used for the detection and quality control of cannabidiol products.

Separation and determination of triadimefon and its metabolites triadimenol enantiomers in fruit puree by supercritical fluid chromatography.

A method was established for the separation and determination of triadimefon and its metabolite triadimenol enantiomer residues in major complementary fruit puree for infants and young children (banana puree, pineapple puree, and grape puree) by supercritical fluid chromatography. After the samples were extracted with acetonitrile and purified with a solid phase extraction cartridge, Acquity Trefoil CEL2 chiral chromatographic column was adopted for separation, and gradient elution was conducted at the flow rate of 1.0 ml/min under the mobile phase of supercritical carbon dioxide - 0.5% ammonia methanol, the detection wavelength was 220 nm and quantification was conducted with the external standard method. The limits of quantitation of triadimefon and triadimenol enantiomers were both 0.05 mg/kg, the linear ranges were 0.5-50 mg/L, and the linear correlation coefficients were greater than 0.9993. The recoveries in the spiked samples at 0.05, 0.2, and 3.0 mg/kg were from 80.1 to 106%, and the relative standard deviation reached 3.3-7.6%. The method is efficient, rapid, reproducible, and environmentally friendly, enabling accurate analysis of pesticide enantiomers, which can detect the enantiomer residues of triadimefon and its metabolite triadimenol in major complementary fruit puree for infants and young children.

Quantitative comparison and rapid discrimination of Panax notoginseng powder and Caulis clematidis armandii using NMR combined with pattern recognition.

BACKGROUND: The market demand for Panax notoginseng (P. notoginseng) is growing rapidly because of its useful properties in food and medicine. However, the frequent adulteration of P. notoginseng seriously affects the health of consumers and is a great challenge to food safety. In this study, low- and high-field nuclear magnetic resonance (LF/HF-NMR) were applied to detect the transverse relaxation distribution of P. notoginseng contaminated with different ratios of Caulis clematidis armandii (CCA) and the components in P. notoginseng and CCA, respectively. RESULTS: Fifty-seven kinds of major and minor components in P. notoginseng and CCA were identified and quantified from their high-resolution NMR spectra, and there were significant differences in ginsenosides, sucrose, and glucose between P. notoginseng and CCA. Furthermore, the partial least squares regression analysis results indicated that LF-NMR parameters (T21 and S21 ) changed linearly as the ratio of CCA increased, and these changes were attributed to the variations in polysaccharide and sucrose in adulterated P. notoginseng. CONCLUSION: In the relaxation time-based pattern recognition models, the authentic P. notoginseng powder could be classified with 100% accuracy from adulterated P. notoginseng when the adulteration ratio was greater than 30%, demonstrating the possibility of LF-NMR, in combination with pattern recognition, for rapid discrimination of food authenticity. © 2022 Society of Chemical Industry.

Research method of rapid determination of chiral pesticide fenpropathrin enantiomers in fruit and vegetable puree by supercritical fluid chromatography.

A method is first established for the separation and determination of fenpropathrin enantiomer residues in apple puree, strawberry puree, and tomato puree considered a supplementary food for infants by supercritical fluid chromatography. After the sample was extracted with acetonitrile and cleaned up by a solid-phase extraction column, then it was separated by a CHIRALPAK AD-3 chiral column with gradient elution at a flow rate of 1.5 mL/min using methanol and supercritical carbon dioxide as the mobile phase, detected by ultraviolet detector at 230 nm wavelength and quantified with the external standard method. The limits of quantification of the two fenpropathrin enantiomers were both 0.2 mg/kg, the linear ranges were 1.0-20.0 mg/L with linear correlation coefficients greater than 0.9992, the recoveries in the spiked samples at 0.2, 0.4 and 2.0 mg/kg were from 80.6 to 105%, and the relative standard deviation reached 2.6-7.7%. This method has the advantages of convenient operation, good resolution, and environmental protection, which can satisfy the requirement of determination for fenpropathrin enantiomer residues in fruit and vegetable puree as supplementary food for infants.

Analytical research on the separation and residue of chiral pesticide triadimenol in fruit and vegetable puree.

In this paper, a method for the separation of triadimenol stereoisomers using ultra-performance convergence chromatography and an analytical method for the determination of triadimenol stereoisomer residues in pumpkin puree, apple puree, and tomato puree as a supplement for infants are established. Test samples were extracted with acetonitrile and successively purified with graphitized carbon black and Florisil column. Afterward, Acquity Trefoil AMY1 column was adopted for chiral separation of chromatographic column, and gradient elute was carried out with supercritical carbon dioxide-methanol as the mobile phase and with external standard method for quantitation. Results showed that the linearly dependent coefficient of the four kinds of triadimenol stereoisomers within 1.0 and 50 mg/L was greater than 0.9997, and the limit of quantitation of the four kinds of triadimenol stereoisomers was 0.05 mg/kg. Recovery experiment was carried out within 0.05 and 1.0 mg/kg scope, the recoveries were 81.0-107％, and the relative standard deviation was 2.3-7.6%. This method implemented the separation of triadimenol stereoisomers and its residue test in pumpkin puree, apple puree, and tomato puree as a supplement for infants, and it can provide reliable technical support for the analysis of pesticide residue and assessment of product quality.

Determination of calcium ß­hydroxy-ß-methylbutyrate content in special medical formula food and sports nutrition food by solid-phase extraction and high performance liquid chromatography.

Calcium ß­hydroxy-ß-methylbutyrate (CaHMB) can promote muscle growth, prevent muscle atrophy, and enhance immunity, therefore, it is widely used as a nutritional supplement in special medical formula food and sports nutrition food. Many methods for the detection of CaHMB have been reported, but the pretreatment method for these reported literatures directly involves extraction using hydrochloric acid solution, without any purification steps. A method for accurately determining CaHMB in special medical formula food and sports nutrition food was established for the first time using solid-phase extraction (SPE) purification and high-performance liquid chromatography method (HPLC). The samples were extracted and precipitated protein using methanol-water solution, purified using SPE method and analyzed by HPLC on diode array detector (DAD) mode under external standard method. The method obtained excellent calibration linearity (r2>0.9993) and a satisfactory analysis of the targeted compound, which were evaluated with calibration standards over the range of 0.020-2.00 mg/mL. The limit of quantifications (LOQs), which defined as the lowest spiking level, were set at 0.4 g/100 g (special medical formula food) and 1.0 g/100 g (sports nutrition food). The average recoveries were within 92.9-104% for the analytes, and the relative standard deviations (RSDs) were below 3.93%, measured at low, medium, and high concentrations. Moreover, the positive sample analysis results indicated that CaHMB was detected on 10 real special medical formula food and sports nutrition food products, the contents of which were generally consistent with their labeled values, with measured values ranging from 97.1 % to 119 % of the labeled values. These results suggested that the developed highly sensitive and specific method is highly feasible for monitoring of the target analyte in special medical formula food and sports nutrition food.

Towards generalizable food source identification: An explainable deep learning approach to rice authentication employing stable isotope and elemental marker analysis.

In addressing the generalization issue faced by data-driven methods in food origin traceability, especially when encountering diverse input variable sets, such as elemental contents (C, N, S), stable isotopes (C, N, S, H and O) and 43 elements measured under varying laboratory conditions. We introduce an innovative, versatile deep learning-based framework incorporating explainable analysis, adept at determining feature importance through learned neuron weights. Our proposed framework, validated using three rice sample batches from four Asian countries, totaling 354 instances, exhibited exceptional identification accuracy of up to 97%, surpassing traditional reference methods like decision tree and support vector machine. The adaptable methodological system accommodates various combinations of traceability indicators, facilitating seamless replication and extensive applicability. This groundbreaking solution effectively tackles generalization challenges arising from disparate variable sets across distinct data batches, paving the way for enhanced food origin traceability in real-world applications.

Origin tracing and adulteration identification of bird's nest by high- and low-field NMR combined with pattern recognition.

Edible bird's nest (EBN) is a high-value health food with various nutrients and bioactive components. With increasing demand for EBN, they are often adulterated with cheaper ingredients or falsely labeled by the origin information, thus harming consumer interests. In this study, high- and low-field nuclear magnetic resonance (HF/LF-NMR) technology combined with multivariate statistical analysis was used to identify the geographical marker of EBN from different origins and authenticate the adulterated EBN with various adulterants at different adulteration rates. Authentic EBN samples from Malaysia were used to simulate adulteration using gelatin (GL), agar (AG) and starch (ST) at 10 %, 20 %, 40 %, 60 %, 80 %, and 100 % w/w, respectively. The results showed significant differences in composition among EBN from different origins, with isocaproate and citric acid serving as geographical markers for Malaysia and Vietnam, respectively. Leucine, glutamic acid, and N-acetylglycoprotein serving as geographical markers for Indonesia. In addition, PLS model further verified the accuracy of origin identification of EBN. The LF-NMR results of adulteration EBN showed a linear correlation between the transverse relaxation (T2, S2) and the adulterated ratio. The OPLS-DA based on T2 spectra could accurately identify authentic EBN from adulterated with GL, AG and ST at 40 %, 20 %, and 20 %, respectively. Fisher discrimination model was able to differentiate at 20 %, 20 %, and 40 %, respectively. These results show that the 1H NMR combined with multivariate statistical analysis method could be a potential tool for the detection of origin and adulteration of EBN.

Target-responsive "sweet" hydrogel with glucometer readout for portable and quantitative detection of non-glucose targets.

Portable devices with the advantages of rapid, on-site, user-friendly, and cost-effective assessment are widely applied in daily life. However, only a limited number of quantitative portable devices are commercially available, among which the personal glucose meter (PGM) is the most successful example and has been the most widely used. However, PGMs can detect only blood glucose as the unique target. Here we describe a novel design that combines a glucoamylase-trapped aptamer-cross-linked hydrogel with a PGM for portable and quantitative detection of non-glucose targets. Upon target introduction, the hydrogel collapses to release glucoamylase, which catalyzes the hydrolysis of amylose to produce a large amount of glucose for quantitative readout by the PGM. With the advantages of low cost, rapidity, portability, and ease of use, the method reported here has the potential to be used by the public for portable and quantitative detection of a wide range of non-glucose targets.

[Simultaneous determination of 28 fragrance components in cosmetics by gas chromatography-tandem mass spectrometry].

With the increasing number of cosmetic products, their flavor and fragrance components are receiving greater and greater attention. Establishing an analytical method of determining these components in cosmetics is one of the most effective measures to eliminate consumers' concerns. In this study, a method for the simultaneous determination of 28 fragrance residues in cosmetics by gas chromatography-tandem mass spectrometry (GC-MS/MS) was developed. The samples were extracted using methanol and those containing more oil and grease were purified using a neutral alumina solid-phase extraction column, whereas those with more complex compositions were purified by QuEChERS. The analytes in the samples were measured by GC-MS/MS, characterized using their retention times and characteristic ion pairs, and quantified with an external standard. The respective limits of detection (LODs, S/N=3) and quantification (LOQs, S/N>10) of the compounds were in the ranges 2-20 and 5-50 µg/kg. The linearities of the concentration curves of the 28 substances were good in the ranges 1-100, 2-200, 4-200, and 10-1000 µg/L, and the correlation coefficients of the quantitative ion pairs were >0.999. Twenty-eight fragrances were added to blank samples at spiked levels of 50-500 µg/kg, and the recoveries ranged from 71.3% to 120.4%, with RSDs of 1.5%-14.6%. The method could be applied in the determination of fragrances in cosmetics because it was simple, sensitive, and stable and could effectively exclude the interferences of complex matrices. The method was used to determine the fragrance components in 16 cosmetic products, and some fragrance components were detected in 12 samples. Increased attention should be paid to the safeties of fragrances and flavors used in cosmetics.

Determination of sugars and sugar alcohols in infant formula by high performance liquid chromatography with evaporative light-scattering detector.

A method was established for the simultaneous determination of five sugars (fructose, glucose, sucrose, lactose, maltose) and five sugar alcohols (erythritol, xylitol, sorbitol, mannitol, maltitol) in infant formula by high performance liquid chromatography-evaporative light scattering detector. After the samples were extracted with acetonitrile-water solution, precipitated by acetic acid, and purified with solid phase extraction cartridge, ALLChrom Rocksil Carbohydrate ES column was adopted for separation, and isocratic elution was conducted at the flow rate of 1.0 mL/min with acetonitrile-0.04 % ammonia solution as the mobile phase. The analytes were detected by an evaporative light-scattering detector, and quantified by external standard method. The linear ranges of the 10 components were 0.04-4.0 g/L with the correlation coefficients greater than 0.999, and the limits of quantification (S/N = 10) of the method were 0.08-0.4 g/100 g. The relative standard deviation of the lactose parallel samples reached 1.29 %, and the recoveries of the other 9 components ranged from 80.4 % to 99.4 % with the relative standard deviation of 2.8 %-7.1 %. The method performs well in sensitivity and separation, which is suitable for the simultaneous quantitative determination of sugars and sugar alcohols in infant formula.

Diversity and Antimicrobial Activity of Intestinal Fungi from Three Species of Coral Reef Fish.

Although intestinal microbiota play crucial roles in fish digestion and health, little is known about intestinal fungi in fish. This study investigated the intestinal fungal diversity of three coral reef fish (Lates calcarifer, Trachinotus blochii, and Lutjanus argentimaculatus) from the South China Sea using a culturable method. A total of 387 isolates were recovered and identified by sequencing their internal transcribed spacer sequences, belonging to 29 known fungal species. The similarity of fungal communities in the intestines of the three fish verified that the fungal colonization might be influenced by their surrounding environments. Furthermore, the fungal communities in different intestines of some fish were significantly different, and the number of yeasts in the hindgut was less than that in fore- and mid-intestines, suggesting that the distribution of fungi in fishes' intestines may be related to the physiological functions of various intestinal segments. In addition, 51.4% of tested fungal isolates exhibited antimicrobial activity against at least one marine pathogenic microorganism. Notably, isolate Aureobasidium pullulans SCAU243 exhibited strong antifungal activity against Aspergillus versicolor, and isolate Schizophyllum commune SCAU255 displayed extensive antimicrobial activity against four marine pathogenic microorganisms. This study contributed to our understanding of intestinal fungi in coral reef fish and further increased the library of fungi available for natural bioactive product screening.

Simultaneous Determination of 147 Pesticide Residues in Traditional Chinese Medicines by GC-MS/MS.

Determination of pesticide residues remains a challenge in traditional Chinese medicines in which complex compounds may interfere with analysis signals. This study reports the development of a simple, effective, and high-throughput method combining gas chromatography-tandem mass spectrometry (GC-MS/MS) with either QuEChERS or solid phase extraction (SPE) to determine 147 pesticide residues in traditional Chinese medicines simultaneously. In SPE, the mixture of n-hexane and ethyl acetate (1:1, v/v) was selected to extract 147 pesticides in honeysuckle, and the extracted pesticides were determined by GC-MS/MS. The limits of detection for all pesticides were within 0.01-0.05 mg/kg. The recoveries were within 70-120% and the relative standard deviations were below 20% for over 90% pesticides. The coefficients of determination were up to 0.999 for the linearity between MS signals and different concentrations of pesticides (20-200 ng/mL). The analytical performance was confirmed in determining pesticide residues in dried tangerine peel. SPE achieved comparable recoveries for all pesticides compared to the QuEChERS method.

Untargeted Metabolomics Analysis Revealed the Difference of Component and Geographical Indication Markers of Panax notoginseng in Different Production Areas.

Panax notoginseng (P. notoginseng) has excellent medicinal and food dual-use characteristics. However, P. notoginseng with a unique origin label has become the target of fraud because of people confusing or hiding its origin. In this study, an untargeted nuclear magnetic resonance (NMR)-based metabolomics approach was used to discriminate the geographical origins of P. notoginseng from four major producing areas in China. Fifty-two components, including various saccharides, amino acids, saponins, organic acids, and alcohols, were identified and quantified through the NMR spectrum, and the area-specific geographical identification components were further screened. P. notoginseng from Yunnan had strong hypoglycemic and cardiovascular protective effects due to its high acetic acid, dopamine, and serine content, while P. notoginseng from Sichuan was more beneficial for diseases of the nervous system because of its high content of fumarate. P. notoginseng from Guizhou and Tibet had high contents of malic acid, notoginsenoside R1, and amino acids. Our results can help to distinguish the geographical origin of P. notoginseng and are readily available for nutritional recommendations in human consumption.

[Determination and level investigation of 45 hormones in edible bird's nest by solid phase extraction-high performance liquid chromatography-tandem mass spectrometry].

Edible bird's nests (EBNs), a food of animal origin, are temporary nests built by swiftlets to foster their offspring. As EBNs and their products are widely accepted by consumers, the safety of hormones in EBNs has also received increasing attention. The establishment of a method for hormone analysis in EBNs and the investigation of hormone levels based on the analytical method are the most effective measures to eliminate any safety concerns. In this study, a multi-residue method was developed for the simultaneous determination of 45 hormones in EBNs, including estrogens, progesterones, androgens, and cortical hormones. EBN samples (1.0 g) were weighed into 50 mL polypropylene centrifuge tubes and mixed with 8 mL of pure water. Then, the samples were extracted twice with 15 mL of acetonitrile and ethyl acetate (1∶1, v/v) under ultrasonic-assisted conditions for 30 min, and the protein in the EBN samples was precipitated at 4000 r/min for 5 min. The clear supernatants were loaded onto a hydrophilic-lipophilic balanced (HLB) SPE column, which was previously activated with methanol (3 mL) followed by pure water (3 mL). The cartridge was washed with 3 mL of pure water and 3 mL of 50% methanol aqueous solution. The hormones were eluted with 3 mL of methanol. A rapid analysis was performed using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The hormones in the extracting solution were separated on a Phenomenex Kinetex C18 column (100 mm×2.1 mm, 2.6 µm) and eluted by gradient elution with acetonitrile-0.1% formic acid aqueous solution (ESI+) or acetonitrile-water (ESI-). Qualitative and quantitative analyses were performed using the multiple reaction monitoring (MRM) mode. The HPLC-MS/MS results showed good linearity in the range of 0.20-20.0 µg/L with correlation coefficients (R2) ≥0.9990. For the 45 hormones, the limits of detection (LODs, S/N≥3) were 0.04-0.70 µg/kg and the limits of quantification (LOQs, S/N≥10) were 0.16-2.00 µg/kg. The recoveries of five hormones, namely, fluorometholone, budesonide, aldosterone, fluocinonide, and ethinylestradiol, were 40.2%-63.6%. Owing to the low recoveries, this method might be suitable only for qualitative testing of the five hormones. The recoveries of the other 40 target analytes were between 72.2% and 112.3% at spiked levels of 2.0, 4.0, and 20.0 µg/kg with relative standard deviations (RSDs) of 2.5%-11.6%. The method is characterized by easy operation, rapidness, high precision, and high sensitivity for the 40 compounds. Thus, this method is suitable for determination of the 40 hormones from EBNs for qualitative testing and quantitation. The proposed method was used to analyze 1021 EBN samples from Malaysia, Indonesia, Thailand, and Vietnam from 2017 to 2021. Only three hormones, progesterone, boldenone, and androstenedione, were identified in the EBN samples, while the levels of the other hormones were lower than their individual LODs. The detected rates of progesterone, boldenone, and androstenedione were 100%, 79%, and 89%, respectively. The contents of progesterone, boldenone, and androstenedione in the EBN samples were 0.097-3.58 µg/kg, 0-0.096 µg/kg and 0-1.77 µg/kg, respectively. The levels of hormones in EBNs were compared with those in eggs, pure milk, and dairy products, which were also animal-derived foods. Androstenedione was detected in all egg samples monitored, and its content was higher than that in EBN samples, pure milk, and dairy products. The content of boldenone was similar among the four products investigated in this study. Based on risk assessment using progesterone, the dietary intake was found to be 3.54 µg/d from milk >1.09 µg/d from eggs >0.0030 µg/d from EBNs. The results showed that the levels of hormones in EBNs were much lower than those in eggs, milk, and dairy products for daily consumption. Based on this investigation, the health effect of the hormones in EBNs may be insignificant.

[Determination of dazomet and its metabolite methyl isothiocyanate residues in plant-derived foods by gas chromatography-triple quadrupole mass spectrometry].

Dazomet is a kind of crystal solid that is stable at room temperature and acts as a fumigant. It is commonly used to control soil fungi, as an insecticide, and in sterilization and weeding. It can effectively kill root-knot nematodes, soil pests, weeds, and many soil-borne disease-causing organisms, to provide clean and healthy soil. Dazomet slowly decomposes and releases methyl isothiocyanate, methylamine, carbon disulfide, and hydrogen sulfide in acidic soil, and diffuses upward through the spaces in the soil to kill contact organisms. When agricultural crops are planted in soil treated with cotton wool, the residues in the grown crop can cause harm to human body when consumed. To ensure the quality and safety of food crops, it is important to develop a detection method for dazomet and its metabolites in plant-derived foods. Hence, in this study, a rapid and simultaneous determination method was developed for dazomet and its metabolite methyl isothiocyanate residues in plant-derived foods by gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS). The sample pretreatment and chromatographic conditions were optimized in the experiment. Subsequently, dazomet and its metabolite methyl isothiocyanate residues in vegetables, fruits, grains, nuts, tea, and spices were extracted with ethyl acetate, and purified using graphitized carbon, a primary-secondary amine, stearyl-bonded silica gel, and anhydrous magnesium sulfate as dispersive solid-phase extraction sorbents. After centrifugation and filtration, the target compounds were analyzed in the multiple reaction monitoring (MRM) mode by GC-MS/MS, and quantified by matrix matching external standard method. The matrix effects of the samples were also evaluated. The matrix effect was found to be in the range of 2.5% to 13.6% for methyl isothiocyanate in 16 matrices. As this matrix effect was weak, there was no need for compensatory measures. In contrast, the matrix effect of dazomet in 16 matrices was in the range of 240.3% to 331.2%. This matrix effect was strong and required compensation. Finally, a matrix matching calibration method was used to compensate the matrix effects. The relative matrix effects of other tested substrates were analyzed using lettuce as the representative substrate; it was found that all showed weak matrix effects. Therefore, the use of lettuce as a representative matrix to prepare a matrix standard curve can effectively correct the matrix effects of dazomet and methyl isothiocyanate in other substrates. Under the optimal conditions, the calibration curves were linear in the range of 0.005-1 mg/L with correlation coefficients higher than 0.99. Recovery tests were conducted by adding mixed standards to blank samples at four levels. The recoveries were in the range of 74.2%-117.2% with relative standard deviations (RSDs, n=6) of 2.8%-9.0%. The limits of quantification (LOQs) of dazomet and methyl isothiocyanate were 0.01 mg/kg. The accuracy and precision of this method met the requirements of pesticide residue determination. The established method was used to detect dazomet and its metabolite methyl isothiocyanate residues in six samples of Chinese cabbage, Chinese chives, cowpea, lettuce, eggplant, ginger, celery, potato, orange, kiwifruit, tomato, chili, rice, tea, almond, and Cuminum cyminum L. in the laboratory, and nothing was detected. The method is simple, rapid, and sensitive; overcomes the shortcomings of existing methods that require two pretreatment steps and two sets of equipment; and meets the requirements for the detection of dazomet and its metabolite methyl isothiocyanate residues in plant-derived foods.

Suspect screening and untargeted analysis of veterinary drugs in food by LC-HRMS: Application of background exclusion-dependent acquisition for retrospective analysis of unknown xenobiotics.

The presence of veterinary drug and pesticide residues in food products pose considerable threats to human health. Monitoring of these residues in food is mainly carried out using targeted analysis by triple quadrupole mass spectrometry. However, these methods are not suitable for suspect screening and untargeted analysis of unknowns. The main objectives of this study were to develop a new high-resolution mass spectrometry (HRMS)-based analytical strategy for retrospective analysis of suspect and unknown xenobiotics and to evaluate its performance in the tentative identification of 48 veterinary drugs as "unknowns" spiked in a pork sample. In the analysis, a newly developed background exclusion data-dependent acquisition (BE-DDA) technique was employed to trigger the product ion (MS/MS) spectral acquisition of the "unknowns", and an in-house precise-and-thorough background-subtraction (PATBS) technique was applied to detect these "unknowns". Results showed that untargeted data mining of the acquired LC-MS dataset by PATBS was able to find all the 48 veterinary drugs and 46 of them were triggered by BE-DDA to generate accurate MS/MS spectra. The dataset of recorded accurate full-scan mass and MS/MS spectra of all the xenobiotics of the test pork sample is defined as the xenobiotics profile. Searching the xenobiotic profile of the test pork sample using mass spectral data of selected veterinary drugs (as suspects) from the mzCloud spectral library led to the correct hits. Searching against the mzCloud spectral library using the mass spectral data of selected individual veterinary drugs (as unknowns) from the xenobiotics profile tentatively confirmed their identities. In contrast, analysis of the same sample using ion intensity-data dependent acquisition only recorded the MS/MS spectra for 34 veterinary drugs. In addition, a data independent acquisition method enabled the acquisition of the fragment spectra for 44 veterinary drugs, but their spectral data displayed only one or a few true product ions of individual analytes of interest along with many fragments from coeluted biological components and background noises. This study demonstrates that this analytical strategy has a potential to become a practical tool for the retrospective suspect screening and untargeted analysis of unknown xenobiotics in a biological sample such as veterinary drugs and pesticides in food products.

[Determination of nine N-nitrosamines in animal derived foods by QuEChERS-isotope dilution combined with gas chromatography-tandem mass spectrometry].

In this study, a comprehensive analytical method based on gas chromatography-tandem mass spectrometry (GC-MS/MS) was developed for the determination of nine N-nitrosamines in animal derived foods. There are many kinds of N-nitrosamines in foods that are harmful to human health. However, the national standard GB 5009.26-2016 pertains only to the detection of N-dimethylnitrosamine; there are many drawbacks of this method, such as complicated sample preparation, low recovery rate, and poor reproducibility. Hence, it is of practical significance to establish a method for the simultaneous determination of a variety of N-nitrosamines. The optimal extraction conditions for the developed method were as follows: 10.0 g aliquots of the sample were placed in a 50 mL centrifuge tube, followed by the addition of 10 mL acetonitrile and 200 µL internal working standard solutions. After 30 min of freezing treatment, 4 g magnesium sulfate and 1 g sodium chloride were added for dehydration, and the tube was centrifuged at 9000 r/min for 5 min. After vortex centrifugation, 5 mL of the clear supernatant was purified using 150 mg polystyrene divinylbenzene (PLS-A). The purified extracts were dewatered using 1.6 g MgSO4 and 0.4 g NaCl, and then filtered through a 0.22 µm membrane filter unit prior to GC-MS/MS analysis. Temperature-programmed was applied at an initial temperature of 50 ℃. After 0.16 min, the temperature was raised to 220 ℃ at the rate of 900 ℃/min for 5 min. N-Nitrosamines were separated on an HP-Innowax column (30 m×0.25 mm×0.25 µm). Identification and quantification were achieved using an electron impact ion (EI) source in positive ion mode with multiple reaction monitoring (MRM). The internal standard method was used to quantify the N-nitrosamines. Under the optimal conditions, the correlation coefficients of the standard calibration curves were not less than 0.99 in the range of 0.1-50.0 µg/L. The limits of detection were 0.03-0.30 µg/kg (S/N=3), and limits of quantification were 0.15-1.00 µg/kg (S/N=10). At spiked levels of 0.5, 1.0, and 3.0 µg/kg, the average recoveries of N-nitrosamines in spiked samples ranged from 80.4% to 98.5%, with relative standard deviations between 2.41% and 12.50%. This method was used to determine animal derived food products, except N-itrosomethylethylamine and N-nitrosomorpholine, others were founded. The results showed that N-nitrosamines levels in salted aquatic products were generally higher than those of the other samples. The method established in this study is simple to operate, and it does not require any time-consuming distillation extraction. Furthermore, there is minimal consumption of samples and reagents; consequently, the experiment cost is reduced, and the method is environmentally friendly. This method has theoretical and practical significance for the control of N-nitrosamines residues in animal derived foods, establishment of detection standards, and corresponding management measures.

[Separation and determination of clenbuterol enantiomers by ultra-performance convergence chromatography].

Clenbuterol enantiomers differ greatly in their bioactivities. By optimizing the conditions for chromatographic separation and method validation, ultra-performance convergence chromatography (UPC2) was adopted to separate the enantiomers of clenbuterol. Standard solutions of (+)-clenbuterol and (-)-clenbuterol were stored at -18 ℃ for 1, 3, 5, 7, 14, 30, and 60 d, and then, their stability was monitored. The impacts of different chromatographic columns, cosolvents, system backpressure, and chromatographic column temperature on the separation of the two enantiomers were investigated. Acquity Trefoil AMY1 (150 mm×3.0 mm, 2.5 µm) was used for separation, and CO2-0.5% (v/v) ammonium acetate was used as the mobile phase. Gradient elution at a flow rate of 2.0 mL/min was adopted. The detection wavelength was set to 241 nm, and the injection volume was set to 10 µL. The backpressure was set to 13.8 MPa, and the column temperature was maintained at 40 ℃. The two enantiomers showed good linear relationships in the range of 1.0 to 20.0 mg/L with correlation coefficients greater than 0.9997. The limits of detection (LODs, S/N=3) of (+)-clenbuterol and (-)-clenbuterol were both 0.5 mg/L. The relative standard deviation (RSD, n=6) for the peak area of the 10.0 mg/L mixed standard working solution with six replicate injections ranged from 0.65% to 0.76%. The effectiveness and practicability of this method were demonstrated by using it to detect standard clenbuterol racemate. The (+)-clenbuterol and (-)-clenbuterol contents were 5.6 mg/L and 5.5 mg/L, respectively, in the standard clenbuterol racemates, as determined by the external standard method of quantification. The detection results suggested that the content ratio of (+)-clenbuterol and (-)-clenbuterol was close to 1.02∶1.00, which is consistent with the literature data. The established method has the advantages of rapid analysis, good separation effect, and low consumption of organic solvents, and it is suitable for the separation of clenbuterol enantiomers. This method can also provide technical support for the separation of other chiral drugs, analysis of the effects of chiral drugs, and assessment of product quality.

[Authenticity identification of manuka honey using liquid chromatography-high resolution mass spectrometry based metabolomic technique].

A liquid chromatography-high resolution mass spectrometry (LC-HRMS) based metabolomic approach for the discrimination of manuka honey from the domestic honey was developed. The results of multivariate data analysis showed significant discrimination of manuka honey from domestic honey. Furthermore, a partial least squares discrimination model was established for honey discrimination, enabling the correct identification of unknown testing samples. A total of 34 highly expressed metabolic markers were obtained from the developed discrimination model, including 3-phenyllactic acid and 4-hydroxybenzoic acid which had both been previously identified as manuka honey markers. The area under the curve of the constructed marker combination reached 0.99. The developed metabolomic approach provides a new method for the quality control of manuka honey.