Performance Liquid Chromatography-quadrupole Time Research Articles

Metabolomic insights into idiopathic xerostomia: The central role of caffeine metabolism in salivary biochemistry

ObjectiveThis study aims to delineate the salivary metabolomic profile of patients with idiopathic xerostomia using untargeted metabolomics techniques, with the goal of addressing the lack of clear diagnostic markers and providing insights into the pathophysiological mechanisms of the condition. DesignIn this observational, cross-sectional study, saliva samples from 33 patients with idiopathic xerostomia and 34 healthy controls were analyzed using Ultra Performance Liquid Chromatography Quadrupole Time of Flight Mass Spectrometry (UPLC-QTOF MS). Metabolomic profiling was complemented by multivariate statistical analysis to differentiate between affected individuals and controls. ResultsMetabolomic analysis delineated a pronounced differentiation between patients with idiopathic xerostomia and healthy controls. A total of 195 metabolites displayed significant differential expression, each with a variable importance in projection (VIP) greater than 1 and a P-value less than 0.05. Pathway enrichment analysis, according to the Kyoto Encyclopedia of Genes and Genomes (KEGG), identified 22 metabolites that participated in 18 distinct metabolic pathways. Among these, the caffeine metabolism pathway, characterized by notable alterations in impact values (VIP, P-value, Log2-fold change, Rich factor), emerged as the most significantly disrupted, underscoring its potential role in the pathophysiology of idiopathic xerostomia (P = 0.0000395). ConclusionsThe salivary metabolomic profiling revealed distinct alterations in idiopathic xerostomia, with a significant reduction in caffeine metabolism pathways, underscoring potential neuropathic involvement. This study advances our understanding of the metabolic alterations in xerostomia, suggesting that salivary metabolomics may offer viable biomarkers for diagnosing and understanding the etiology of idiopathic xerostomia. Future research should focus on therapeutic targeting of these metabolic disturbances and evaluating their reversibility with treatment.

Integrated Transcriptomics and Metabolomics Reveal Key Insights into Iridoid Biosynthesis in Gentiana crassicaulis Seeds during Germination.

Background:Gentiana crassicaulis Duthie ex Burk., a key species used in traditional Chinese medicine for treating rheumatic pain and stroke, contains iridoids as its primary active component. However, the biosynthetic mechanisms underlying iridoid production are not fully understood. Methods: This study focused on iridoid biosynthesis during the germination of G. crassicaulis seeds, integrating metabolomic and transcriptomic analyses to uncover the underlying pathways and key candidate genes. Results: 196,132 unigenes and 10 iridoid compounds were identified through RNA-seq and ultra performance liquid chromatography-quadrupole time of flight-mass spectrometer (UPLC-Q-TOF-MS), respectively. The intersection of results from Pearson correlation analysis and weighted gene co-expression network analysis (WGCNA) revealed a significant correlation between 26 genes and iridoid levels, suggesting their potential role in the iridoid metabolism. Notably, six highly expressed candidate genes (DL7H, SLS, CYP76, CYP72A2, CYP84A1, and 13-LOX3) and five iridoids (loganic acid, sweroside, swertiamarin, gentiopicroside, and 6'-O-β-D-glucosyl-gentiopicroside) responded to methyl jasmonate stimulation in G. crassicaulis seedlings. Conclusions: by combining the known functions of candidate gene families, It is hypothesized that the CYP716 and LOX families exert indirect influences on iridoid metabolism, while the CYP71, CYP81, CYP72, CYP76, CYP710 families, 2OG-FeII family, and the glucosyltransferase family are likely to play direct roles in the biosynthetic transformations of the five iridoids. This study provides a theoretical basis for further functional gene validation and metabolic engineering aimed at enhancing iridoid production. The insights gained could lead to improved iridoid production efficiency in medicinal plants, ultimately benefiting the quality and efficacy of medicinal materials.

Differential Chemical Components Analysis of Periplocae Cortex, Lycii Cortex, and Acanthopanacis Cortex Based on Mass Spectrometry Data and Chemometrics.

Background:Periplocae Cortex (PC), Acanthopanacis Cortex (AC), and Lycii Cortex (LC), as traditional Chinese medicines, are all dried root bark, presented in a roll, light and brittle, easy to break, have a fragrant scent, etc. Due to their similar appearances, it is tough to distinguish them, and they are often confused and adulterated in markets and clinical applications. To realize the identification and quality control of three herbs, in this paper, Ultra Performance Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry Expression (UHPLC-QTOF-MSE) combined with chemometric analysis was used to explore the different chemical compositions. Methods: LC, AC, and PC were analyzed by UHPLC-QTOF-MSE, and the quantized MS data combined with Principal Component Analysis (PCA) and Partial Least Squares Discriminant Analysis (PLS-DA) were used to explore the different chemical compositions with Variable Importance Projection (VIP) > 1.0. Further, the different chemical compositions were identified according to the chemical standard substances, related literature, and databases. Results: AC, PC, and LC can be obviously distinguished in PCA and PLS-DA analysis with the VIP of 2661 ions > 1.0. We preliminarily identified 17 differential chemical constituents in AC, PC, and LC with significant differences (p < 0.01) and VIP > 1.0; for example, Lycium B and Periploside H2 are LC and PC's proprietary ingredients, respectively, and 2-Hydroxy-4-methoxybenzaldehyde, Periplocoside C, and 3,5-Di-O-caffeoylquinic acid are the shared components of the three herbs. Conclusions: UHPLC-QTOF-MSE combined with chemometric analysis is conducive to exploring the differential chemical compositions of three herbs. Moreover, the proprietary ingredients, Lycium B (LC) and Periploside H2 (PC), are beneficial in strengthening the quality control of AC, PC, and LC. In addition, limits on the content of shared components can be set to enhance the quality control of LC, PC, and AC.

Detection of the synthetic cathinone N,N-dimethylpentylone in seized samples from prisons

Drug use is prevalent in prisons with drugs associated with depressant effects found to be more prevalent than stimulants. Synthetic cathinones (SCats; often sold as “bath salts”, “ecstasy”, “molly”, and “monkey dust”) are the second largest category of new psychoactive substances (NPS) currently monitored by the European Monitoring Centre for Drugs and Drug Addiction (EMCDDA) and are commonly used as substitutes for regulated stimulants, such as amphetamine, cocaine, and MDMA. N,N-dimethylpentylone (also known as dimethylpentylone, dipentylone, and bk-DMBDP) was detected for the first time in the Scottish prisons in seven powder samples seized between January and July 2023. Samples were analyzed using gas chromatography-mass spectrometry (GC-MS), ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-QToF-MS), and nuclear magnetic resonance imaging (NMR). Dimethylpentylone was detected alongside other drugs in four samples, including the novel benzodiazepine desalkylgidazepam (bromonordiazepam) and the synthetic cannabinoid receptor agonists (SCRAs) MDMB-INACA and MDMB-4en-PINACA.

New insights into the chemistry and anticancer activity of Ophiocordyceps xuefengensis

Ophiocordyceps xuefengensis (O. xuefengensis), the sister taxon of Ophiocordyceps sinensis (O. sinensis), is consumed as a “tonic food” due to its health benefits. However, little is known regarding the chemistry and bioactivity of O. xuefengensis. In this study, we characterized 80 indole-based alkaloids in the ethyl acetate fraction of O. xuefengensis by high performance liquid chromatography-quadrupole time of flight mass spectrometry (HPLC–Q–TOF–MS/MS), of which 54 indole-based alkaloids were identified as possibly new compounds. Furthermore, 29 of these compounds were established as potential anti-cancer compounds by ligand fishing combined with HPLC–Q–TOF–MS/MS. Moreover, molecular docking identified the NH- and OH- groups of these compounds as the key active groups. The present study has expanded the knowledge on the characteristic indole-based alkaloids and anti-cancer activity of O. xuefengensis.

Cai's herbal tea enhances mitochondrial autophagy of type 1 diabetic mellitus β cells through the AMPK/mTOR pathway and alleviates inflammatory response.

This study investigates the therapeutic mechanisms of Cai's Herbal Tea in Type 1 Diabetes Mellitus (T1DM) mice, focusing on its effects on mitochondrial change and autophagy via the AMP-activated protein kinase (AMPK)-mammalian target of rapamycin (mTOR) pathway. The composition of Cai's Herbal Tea was analyzed by Ultra-High Performance Liquid Chromatography-Quadrupole Time of Flight Mass Spectrometry (UHPLC-Q/TOF-MS). C57BL/6 mice and Min6 pancreatic beta cells were divided into control, diabetic mellitus (DM)/high glucose (HG), and treatment groups (low, medium, and high doses of Cai's Tea, and Metformin). Key physiological parameters, pancreatic islet health, Min6 cell morphology, viability, and insulin (INS) secretion were assessed. Small Interfering RNA-AMPK (si-AMPK) was utilized to confirm the pathway involvement. Cai's Herbal Tea improved body weight, pancreatic islet pathological injury, and INS secretion whereas reduced total triglycerides, fasting blood sugar, and Interferon gamma (INF-γ) in T1DM mice, particularly at higher doses. In Min6 cells, Cai's Tea mitigated HG-induced damage and proinflammatory response, enhancing cell viability and INS secretion. Notably, it reduced swelling and improved cristae structure in treated groups of mitochondria and promoted autophagy via the AMPK-mTOR pathway, evidenced by increased LC3II/LC3I and P-AMPK/AMPK ratios, and decreased P-mTOR/mTOR and P62 expressions in pancreatic islet β-cells. Furthermore, these effects were converted by si-AMPK interference. Cai's Herbal Tea exhibits significant therapeutic efficacy in T1DM mice by improving mitochondrial health and inducing autophagy through the AMPK-mTOR pathway in pancreatic islet β-cells. These findings highlight its potential as a therapeutic approach for T1DM management.

Determination of twelve neonicotinoid pesticides in chili using an improved QuEChERS method with UPLC-Q-TOF/MS

In this study, a QuEChERS method based on citrate was developed and utilized for the analysis of twelve neonicotinoid pesticides in fresh red chilies, fresh green chilies, and dried chilies, coupled with ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS). In the sample preparation, acetonitrile containing 1% formic acid was used as the extraction solvent. Anhydrous sodium sulfate replaced the traditional anhydrous magnesium sulfate for water removal, effectively eliminating the issues of salt caking. Graphitized carbon black, octadecyl silica, and primary secondary amine were used as cleaning agents. The method showed good sensitivity, with the limits of quantification below 0.03 mg/kg for fresh chilies and below 0.15 mg/kg for dried chilies. Values of matrix effects ranged from −19.5% to 8.4%, and the recovery was 86.9% - 105.2%. The analytical method provided an effective tool for the high throughput detection of neonicotinoid pesticide residues in multiple chili matrices.

A breakthrough in periodontitis treatment: Revealing the pharmacodynamic substances and mechanisms of Kouqiangjie formula

Ethnopharmacology relevancePeriodontitis, a complex inflammatory disease, significantly affects people's lives. Traditional Chinese multi-herbal formulas, composed of various herbs, exhibit their therapeutic efficacy holistically. Kouqiangjie Formula (KQJF), comprising 12 herbs including Rhizoma smilacis glabrae, Polygonatum sibiricum Delar. ex Redoute, Taraxacum mongolicum Hand.-Mazz, etc., has been clinically proven to effectively treat periodontitis. However, the potential active substances conferring these effects and their mechanisms of action remain unclear. Aim of the studyThe current investigation endeavours to utilize Ultra Performance Liquid Chromatography Quadrupole Time of Flight Mass Spectrometry (UPLC-Q-TOF-MS), network pharmacology, and in vivo animal experiment confirmation to explore the plausible bioactive compounds and operational mechanisms underpinning KQJF's therapeutic impact on periodontitis. Materials and methodsUsing the UPLC-Q-TOF-MS technique, we deciphered the chemical constituents of KQJF. Network pharmacology was employed to earmark key bioactive elements, forecast principal targets, and operational pathways which were later substantiated through molecular docking. Experimental validations were carried out in a periodontitis animal model using a range of techniques, including micro-CT, H&E staining, qRT-PCR, and protein blotting procedures, providing comprehensive verification of our initial assumptions. ResultsUtilizing UPLC-Q-TOF-MS, we characterized 87 individual chemical constituents in KQJF. Network pharmacology revealed that 14 components, including senkyunolide A, glycycoumarin, licoflavonol, glycyrin, senkyunolide I, and senkyunolide H, form the key therapeutic basis of KQJF in targeting periodontitis. Significant targets and pathways were discerned as AKT1, MMP9, JUN, PTGS2, CASP3, TLR4, IL1β, BCL2, PPARG, and pathways such as the TNF signaling pathway, NF-κB signaling pathway, osteoclast differentiation, and Wnt signaling pathway. Molecular docking demonstrated robust binding activity between these crucial targets and the key active ingredients. In vivo experimentation corroborated that, compared with the model group, KQJF significantly ameliorated symptoms and micro-CT imaging parameters of periodontitis in the rat model, down-regulating the expression of AKT1, MMP9, JUN, PTGS2, CASP3, TLR4, and IL1β, while up-regulating the expression of BCL2 and PPARG. ConclusionIn summary, this study has pioneered a comprehensive exploration of the potential therapeutic constituents, targets, and mechanisms of KQJF for periodontitis treatment, adopting a synergistic strategy of “chemical component analysis-network pharmacology screening-in vivo animal experiment validation”. This provides experimental evidence for the clinical application of KQJF and further in-depth research. Additionally, it presents an effective strategy for the research of other Chinese herbal formulations.

ERCC1 which affects lipids metabolism and actin dynamics in coal workers' pneumoconiosis is a candidate biomarker for early warning and diagnosis.

The single-nucleotide polymorphisms of genes related to DNA damage repair and inflammasomes and mutated gene expression in coal workers' pneumoconiosis (CWP) were analysed to identify the risk factors of CWP and potential biomarkers for early warning and diagnosis. Further, mutated gene pathways were analysed based on proteome and metabolome. Han Chinese male subjects were randomly selected and divided into 4 or 5 groups according to the process of CWP. MassARRAY was used to sequence single-nucleotide polymorphism genotypes. Mutated gene expression in plasma was tested using enzyme-linked immunosorbent assay (ELISA). Odds ratios (ORs) and receiver operating characteristic curves (ROC) were calculated. The serum different proteins and metabolites were identified by Ultra Performance Liquid Chromatography Quadrupole time of flight/Mass Spectrum (UPLC-Q-TOF/MS) and analysed using bioinformation software. As CWP progressed, the CC and CA genotypes of ERCC1 rs3212986 decreased and increased significantly, respectively. AA (OR = 3.016) and CA (OR = 2.130) genotypes were identified as risk factors for stage II. ERCC1 significantly decreased in processing of CWP. The cutoff value of ERCC1 was 5.265 pg/ml, with a sensitivity of 90.0% and specificity of 86.7%. ERCC1 had an indirect interaction with activator protein-1 and insulin and its pathways were mainly made with molecules related to lipid metabolism and actin dynamics. ERCC1 is a candidate biomarker for detection and precise intervention in CWP. If it reaches the threshold, workers will change other jobs in time and will not develop and diagnose as pneumoconiosis and will help the employers spend less money. Meanwhile, the signal molecules of ERCC1 pathway could be as a candidate target for drug discovery.

Method development and optimization for dispersive liquid-liquid microextraction factors using the response surface methodology with desirability function for the ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry determination of organic contaminants in water samples: risk and greenness assessment.

A simple, cost effective, and efficient dispersive liquid-liquid microextraction method was developed and optimized for the determination of organic contaminants in different environmental water matrices followed by UHPLC-QTOF-MS analysis. In the preliminary experiments, the univariate optimization approach was used to select tetrachloroethylene and acetonitrile as extraction and disperser solvents, respectively. The significant factors influencing DLLME were screened using full factorial design, and the optimal values for each variable were then derived through further optimization using central composite design with desirability function. The optimal conditions were achieved with 195 μL of tetrachloroethylene as the extraction solvent, 1439 μL of acetonitrile as the disperser solvent, and a sample pH of 5.8. Under these conditions, the method provided detection limits ranging from 0.11-0.48 μg L-1 and recoveries ranging from 23.32-145.43% across all samples. The enrichment factors obtained ranged from 11.66-72.72. The proposed method was then successfully applied in real water samples. Only benzophenone was detected in the concentration range of 0.79-0.88 μg L-1 across all the water samples. The calculated risk quotient resulting from benzophenone exposure in water samples showed a low potential risk to human health and the aquatic ecosystem. The method was also evaluated for its environmental friendliness using various metrics tools such as Analytical Eco-Scale (AES), Green Analytical Procedure Index (GAPI), Analytical GREEnness (AGREE), Analytical Greenness for Sample Preparation (AGREEprep), and Sample Preparation Metric of Sustainability (SPMS). Only AES qualified the method as green while it was considered acceptable and sustainable when assessed using SPMS.

Comparative Metabolomics Revealed Differences in Alkaloids Metabolism Between Morus nigra L. and Morus. alba L. at Different Growth Stages

Mulberry (Morus spp.), a flowering plant in the Moraceae family, is known for the edible fruit, and mulberry leaves have various applications. This experiment utilized ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS) and Progenesis QI software to rapidly identify secondary metabolites in the leaves of Morus nigra L. and Morus alba L.. Orthogonal partial least squares discriminant analysis (OPLS-DA), principal component analysis (PCA) and Hierarchical clustering analysis (HCA) were used to examine the dynamic changes and stage specificity of metabolites, particularly alkaloids, in M. nigra and M. alba leaves during seven primary growth stages. In results, the leaf samples from different growth stages uncovered a total of 182 metabolites from 13 distinct categories. M. nigra had significantly higher levels of metabolites than those in M. alba, according to differences in leaf metabolite accumulation patterns. Multiple statistical analyses revealed that the metabolome of M. nigra and M. alba leaves exhibited a significant interspecific and developmental specific accumulation patterns, with most DNJ alkaloids having the highest content in M. nigra leaves. In different growth stages, the metabolites of M. nigra leaves showed significant stage specificity. During the development stage of small fruit stage, obvious changes occurred in the chemical composition of M. nigra leaves, with a significant increase in DNJ alkaloid metabolites. Overall, this study provides a theoretical reference for the comprehensive development and sustainable utilization of M. nigra resources in Xinjiang.

Characterization of key bitter compounds in Idesia polycarpa var. vestita Diels fruit by sensory-guided fractionation

Idesia polycarpa var. vestita Diels (I. vestita) has become a promising oil crop due to its easily digestible and highly nutritious fruit oil. However, the intense bitter taste of its fruit greatly limits its development and promotion in the food industry. Herein, five key bitter compounds from I. vestita fruit were isolated by sensory-guided fractionation and characterized using ultra-high performance liquid chromatography-quadrupole time of flight-mass spectrometer and nuclear magnetic resonance. The bitter taste of the identified compounds was subsequently validated by threshold tests and computational molecular docking. The bitterness threshold in water of idesin was the lowest (12.051 mg/L), and all bitter substances spontaneously bound to the bitter receptors hTAS2R16 and hTAS2R14, with a stronger affinity for the latter (approximately −6.5 – −9.0 kcal/mol). This is the first systematic study of bitter compounds in I. vestita fruit, providing a scientific basis for revealing the mechanism of bitterness formation and bitterness control.

Electrooxidation of amoxicillin in aqueous solution with graphite electrodes: Optimization of degradation and deciphering of byproducts using HRMS

Contaminants of emerging concern (CECs) such as antibiotics have become a matter of worry in aquatic environments worldwide. Their presence in the environment has been increasing due to the inability of conventional wastewater and water treatments to annihilate them. Hence, attempts have been made to remove CECs using electrochemical oxidation (EO). Present study employed the low cost, active carbon based graphite sheet electrodes as anode and cathode to oxidize and degrade Amoxicillin (AMOX)- a β-lactum thiazolidine antibiotic. Optimization studies found pH 9, 45 mA cm−2, 81 cm2 electrode surface area, 6 mM electrolyte concentration and 60 min treatment time to be optimal for AMOX removal. Studies with varying concentrations of AMOX (20 mg L−1, 30 mg L−1 and 40 mg L−1) found that increase in concentrations of AMOX require higher current densities and treatment time for better TOC removal. High performance liquid chromatography photo diode array (HPLC-PDA) studies found 94% removal for 40 mg L−1 of AMOX at optimal conditions with 90% COD and 46% TOC removal. High resolution mass spectrometry (HRMS) studies using Ultra performance liquid chromatography-quadrupole time of flight-mass spectrometry (UPLC-Q-ToF-MS) identified major degradation mechanisms to be hydroxylation, β-lactum ring cleavage, breakage of thiazolidine ring chain from the aromatic ring and piperazinyl ring formation. The final byproducts of AMOX oxidation were carboxylic acids.

Discovery, isolation and structural identification of alkaloid glycosides in six traditional Chinese medicine such as Coptis chinensis

Alkaloids are important active ingredients occurring in many traditional Chinese medicines, and alkaloid glycosides are one of their existence forms. The introduction of saccharide units improves the water solubility of alkaloid glycosides thus presenting better biological activity.Because of the low content in plants, alkaloid glycosides have been not comprehensively studied. In this study, ultrahigh performance liquid chromatography-quadrupole time of flight-tandem mass spectrometry(UPLC-QTOF-MS/MS) was employed to identify and analyze the alkaloid glycosides in Coptis chinensis, Phellodendron chinense, Menispermum dauricum, Sinomenium acutum, Tinospora sagittata and Stephania tetrandra. The results showed that except Tinospora sagittata, the other five herbal medicines contained alkaloid glycosides. Furthermore, the alkaloid glycosides in each herbal medicine were identified based on UV absorption spectra, quasimolecular ion peaks in MS, fragment ions information in the MS/MS, and previous literature reports. A total of 42 alkaloid glycosides were identified. More alkaloid glycosides were identified in C. chinensis and Menispermum dauricum, and eleven in C. chinensis were potential new compounds. Furthermore, the alkaloid glycosides in the water extract of C. chinensis were coarsely se-parated by macroporous adsorption resin, purified by column chromatography with D151 cation exchange resin, ODS and MCI, combined with semi-preparative high performance liquid chromatography. Two new alkaloid glycosides were obtained, and their structures were identified by mass spectrometry and NMR data as(S)-7-hydroxy-1-(p-hydroxybenzyl)-2,2-N,N-dimethyl-1,2,3,4-tetrahydroisoquinoline-6-O-β-D-glucopyranoside and(S)-N-methyltetrahydropalmatubine-9-O-β-D-glucopyranoside, respectively. This study is of great significance for enriching the information about the chemical composition and the in-depth development of C. chinensis. Meanwhile, it can provide a reference for rapid identification and isolation of alkaloid glycosides from other Chinese herbal medicines.

6-Oxofurostane and (iso)Spirostane Types of Saponins in Smilax sieboldii: UHPLC-QToF-MS/MS and GNPS-Molecular Networking Approach for the Rapid Dereplication and Biodistribution of Specialized Metabolites.

Identifying novel phytochemical secondary metabolites following classical pharmacognostic investigations is tedious and often involves repetitive chromatographic efforts. During the past decade, Ultra-High Performance Liquid Chromatography-Quadrupole Time of Flight-Tandem Mass Spectrometry (UHPLC-QToF-MS/MS), in combination with molecular networking, has been successfully demonstrated for the rapid dereplication of novel natural products in complex mixtures. As a logical application of such innovative tools in botanical research, more than 40 unique 3-oxy-, 3, 6-dioxy-, and 3, 6, 27-trioxy-steroidal saponins were identified in aerial parts and rhizomes of botanically verified Smilax sieboldii. Tandem mass diagnostic fragmentation patterns of aglycones, diosgenin, sarsasapogenin/tigogenin, or laxogenin were critical to establishing the unique nodes belonging to six groups of nineteen unknown steroidal saponins identified in S. sieboldii. Mass fragmentation analysis resulted in the identification of 6-hydroxy sapogenins, believed to be key precursors in the biogenesis of characteristic smilaxins and sieboldins, along with other saponins identified within S. sieboldii. These analytes' relative biodistribution and characteristic molecular networking profiles were established by analyzing the leaf, stem, and root/rhizome of S. sieboldii. Deducing such profiles is anticipated to aid the overall product integrity of botanical dietary supplements while avoiding tedious pharmacognostic investigations and helping identify exogenous components within the finished products.

Determination of multiple mycotoxins in chili powder using cold-induced liquid–liquid extraction and Fe3O4@MWCNTs-NH2 coupled with UPLC-Q-TOF/MS

Food matrix interference is still a big challenge in analyzing multiclass mycotoxins. Herein, a novel cold-induced liquid–liquid extraction-magnetic solid phase extraction (CI-LLE-MSPE) coupled with ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-Q-TOF/MS) method was explored for the simultaneous determination of multiple mycotoxins in chili powders. Fe3O4@MWCNTs-NH2 nanomaterials were prepared and characterized, and the factors affecting the MSPE process were investigated. Based on this, the CI-LLE-MSPE-UPLC-Q-TOF/MS method was established for determining ten mycotoxins in chili powders. The proffered technique eliminated the matrix interference effectively and demonstrated strong linearity (0.5–500 µg/kg, R2 ≥ 0.999), high sensitivity (limit of quantification was 0.5–1.5 µg/kg), and the recovery was 70.6%-111.7%. The extraction process is simpler than conventional methods, as the adsorbent can be separated using magnets, and reusable adsorbents are beneficial in reducing costs. In addition, the method can provide a valuable reference for pretreatment procedures for other complex matrices.

Metabolites profiling of protein enriched oyster mushroom (Pleurotus ostreatus (Jacq.) P. Kumm.) grown on oil palm empty fruit bunch substrate

This study aimed to compare the nutritional composition and metabolites profiles of the Pleurotus ostreatus oyster mushroom grown on oil palm empty fruit bunch (EFB) and rubber wood sawdust (RWS) substrates. The mushrooms were prepared for proximate analysis, beta-glucan content, and metabolites profiling using ultra high performance liquid chromatography-quadrupole time of flights mass spectrometer (UHPLC-QToF). Proximate analysis showed significantly higher crude protein, and beta-glucan contents (p < 0.05) were recovered from the mushrooms grown on EFB substrate as compared to those grown on RWS. Metabolite profiling of hot water extracts showed that three lipids (PE (17:1/0:0), PE (16:1/0:0), and hydroxyoctadecadienoic), two amino acids (acetyl-leucine, and threonic acid), and one polysaccharide (4-O-beta-d-galactopyranosyl-beta-d-xylopyranose) were present at a relatively higher level (maximum fold change >10) in mushrooms grown on EFB. Our results suggest that the metabolites of the mushroom cultivated on EFB substrate were altered. The innovative use of palm oil waste as mushroom cultivation substrate exerts the concept of waste-to-wealth, promoting Sustainable Palm Oil Initiative (SPOI), and enhancing food security through the production of high-protein mushrooms.

Mitochondrial Metabolomics of Sym1-Depleted Yeast Cells Revealed Them to Be Lysine Auxotroph.

Metabolomics has expanded from cellular to subcellular level to elucidate subcellular compartmentalization. By applying isolated mitochondria to metabolome analysis, the hallmark of mitochondrial metabolites has been unraveled, showing compartment-specific distribution and regulation of metabolites. This method was employed in this work to study a mitochondrial inner membrane protein Sym1, whose human ortholog MPV17 is related to mitochondria DNA depletion syndrome. Gas chromatography-mass spectrometry-based metabolic profiling was combined with targeted liquid chromatography-mass spectrometry analysis to cover more metabolites. Furthermore, we applied a workflow employing ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry with a powerful chemometrics platform, focusing on only significantly changed metabolites. This workflow highly reduced the complexity of acquired data without losing metabolites of interest. Consequently, forty-one novel metabolites were identified in addition to the combined method, of which two metabolites, 4-guanidinobutanal and 4-guanidinobutanoate, were identified for the first time in Saccharomyces cerevisiae. With compartment-specific metabolomics, we identified sym1Δ cells as lysine auxotroph. The highly reduced carbamoyl-aspartate and orotic acid indicate a potential role of the mitochondrial inner membrane protein Sym1 in pyrimidine metabolism.

Facile synthesis of triazine-based porous organic polymer for the extraction and determination of nitrofuran metabolites residues from meat samples with ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry

In this work, a novel triazine-based porous organic polymers, TAPT-BPDD, was firstly synthesized by a facile method at room temperature. After characterized by FT-IR, FE-SEM, XRPD, TGA, and nitrogen-sorption experiments, TAPT-BPDD was applied as solid-phase extraction (SPE) adsorbent for the extraction of four trace nitrofuran metabolites (NFMs) from meat samples. The key parameters including the adsorbent dosage, sample pH, type and volume of eluents, type of washing solvents were evaluated in the extraction process. Combined with ultra-high performance liquid chromatography-quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS/MS) analysis, good linear relationship (1–50 µg·kg−1, R2>0.9925) and low limits of detection (LODs, 0.05–0.56 µg·kg−1) were obtained under the optimal conditions. When spiked at different level, the recoveries were in the range of 72.7–111.6%. The adsorption isothermal model and extraction selectivity of TAPT-BPDD were also studied in detail. The results showed that TAPT-BPDD was a kind of promising SPE adsorbent for the enrichment of organics in food samples.

An efficient method for qualitation and quantitation of multi-components of the herbal medicine Qingjin Yiqi Granules

Qingjin Yiqi Granules (QJYQ) is a Traditional Chinese Medicines (TCMs) prescription for the patients with post-COVID-19 condition. It is essential to carry out the quality evaluation of QJYQ. A comprehensive investigation was conducted by establishing deep-learning assisted mass defect filter (deep-learning MDF) mode for qualitative analysis, ultra-high performance liquid chromatography and scheduled multiple reaction monitoring method (UHPLC-sMRM) for precise quantitation to evaluate the quality of QJYQ. Firstly, a deep-learning MDF was used to classify and characterize the whole phytochemical components of QJYQ based on the mass spectrum (MS) data of ultra-high performance liquid chromatography quadrupole time of flight tandem mass spectrometry (UHPLC-Q-TOF/MS). Secondly, the highly sensitive UHPLC-sMRM data-acquisition method was established to quantify the multi-ingredients of QJYQ. Totally, nine major types of phytochemical compounds in QJYQ were intelligently classified and 163 phytochemicals were initially identified. Furthermore, fifty components were rapidly quantified. The comprehensive evaluation strategy established in this study would provide an effective tool for accurately evaluating the quality of QJYQ as a whole.