Quadrupole Orbitrap High-resolution Mass Spectrometry Research Articles

Degradation of non-phthalate plasticisers in soils by liquid chromatography coupled to high resolution mass accuracy spectrometry: A comparison with sterile soil

The market for non-phthalate plasticisers (NPPs), such as adipates, citrates, phosphates, and carboxylates, has grown due to their low toxicity, but no studies have determined their metabolites during degradation processes in soil. This study evaluated the degradation of diethyl hexyl adipate (DEHA) and acetyl triethyl citrate (ATEC) in nonsterile soil, and the results were compared with those obtained from sterile soil, to determine whether microbial activity causes degradation in these NPPs. The analyses of these substances were performed using liquid chromatography coupled with quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap-HRMS). Plasticiser degradation followed a biphasic kinetic model, with a half-life time (DT50) of 9–12 days in non-sterile soil, but very slow in sterile soil (DT50 > 58 days). The degradation of target plasticisers in nonsterile soil resulted in the detection of eight metabolites of DEHA and ATEC, which were not detected in sterile soil. Among them, 4-[(2-ethylhexyl)oxy]-4-oxobutanoic acid (EHOBA), 4-((2-ethylhexyl)oxy)-4-oxo-1-carbonyl hexanoate (EHOCH), triethylcitrate (TEC) and N-acetyl glutamine (NAG) have been detected for the first time in this study. Most of these metabolites were detected after 14 days, increasing their concentration to 21 days, before completely degrading after 45 days. Nevertheless, mono–2-ethyl-5-hydroxyhexyl adipate (MEHHA), mono-2-ethyloxohexyl adipate (MEHOA), EHOCH, and diethyl citrate (DEC) were persistent in non-sterile soil (detected at 45 days) with an LD50 lower than their respective original molecules. Therefore, it is necessary to control the amount of these metabolites derived from the degradation of this type of plasticisers in the soil to avoid health risks, as they can end up in vegetables.

Zilpaterol in bovine liver, meat, heart, and kidney, determined by liquid chromatography–quadrupole-orbitrap high-resolution mass spectrometry

ABSTRACT An analytical method was developed for identification and quantification of zilpaterol in bovine liver, meat, heart, and kidney, using liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS). It was validated in accordance with Commission Implementing Regulation (CIR) EU 2021/808 at six different concentrations, ranging from 0.1 to 5 μg/kg. The mean recoveries ranged from 71% to 99%, while the decision limit (CCα) and detection capability (CCβ) ranges were 0.11–0.12 μg/kg and 0.13–0.15 μg/kg, respectively. The method demonstrated good linearity (R2 > 0.9996) and the limits of detection (LODs) and of quantification (LOQs) were in the range of 0.015–0.061 μg/kg and 0.025–0.091 μg/kg, respectively. Out of 200 samples collected from local markets in Egypt, 17 contained zilpaterol residues. Liver samples revealed the highest detection frequency (26%), followed by meat (6%), at mean concentrations of 2.64 and 1.93 μg/kg, respectively.

Identification of chemical constituents and pharmacokinetic characteristics of Xiaoyan Tuire Granule in rats

Xiaoyan Tuire Granule is a type of Chinese patent medicine that has been proven effective in treating respiratory tract infections. However, while it has been successfully introduced into clinical use, more knowledge is still needed regarding its chemical components and pharmacokinetics. This study investigated the chemical profile in the medicine and rat plasma by ultra high-performance liquid chromatography coupled with Q Exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry (UHPLC-Orbitrap-MS/MS). Subsequently, it developed a validated ultra high-performance liquid chromatography coupled with quadrupole mass spectrometry (UHPLC-MS/MS) method for determining five components in rat plasma after oral administration of Xiaoyan Tuire Granule. As a result, a total of 106 constituents were inferred, including 9 terpenoids, 29 flavonoids, 33 organic acids, 12 phenylpropanoids and 23 other compounds. After administration, 86 compounds were inferred in rat plasma, including 73 prototypes and 13 metabolites. The metabolic pathways were primarily hydrogenation, glucuronic acid conjugation, sulfate conjugation, hydrolysis and methylation. The established method determined the contents of esculetin, esculin, isovitexin, caffeic acid and p-coumaric acid had a good separation, and all the legal verification met the requirements. The pharmacokinetic results indicate that the absorption rate of the five compounds in vivo was rapid, with a Tmax of less than 0.25 h, and the elimination rate was also fast, with a half-time (T1/2) ranging from 1.22 h to 2.19 h. It is worth noting that esculin and esculetin have similar half-time in vivo due to their structural similarities. Among these five compounds, the AUC0-∞ and MRT0-∞ of p-coumaric acid and esculetin were relatively higher, indicating higher exposure and longer residence time of both compounds in vivo. In conclusion, this paper researched the chemical constituents and pharmacokinetics of Xiaoyan Tuire Granule, which provided the reference for further study.

Quantitation of trace polyfunctional thiols in wine by liquid chromatography quadrupole Orbitrap high-resolution mass spectrometry in parallel reaction monitoring

Polyfunctional thiols are key contributors to wine aroma due to their extremely low odor thresholds, and their quantitative analysis remains challenging as a result of their ultratrace concentrations and high reactivity. This work presents the first method based on ultra-high-performance liquid chromatography (UHPLC) coupled to quadrupole Orbitrap high-resolution mass spectrometry (HRMS) in parallel reaction monitoring (PRM) mode for quantifying thiols at nanograms per liter (ng/L) levels in wine. Thiols in wine were derivatized using 4,4′-dithiodipyridine and isolated by liquid-liquid extraction. This protocol allowed rapid sample preparation with minimum labor input and low consumable expenses. Instrumental analysis was conducted using UHPLC–quadrupole Orbitrap HRMS in PRM mode. Twenty thiol analytes, including literature-known, recently identified, and novel thiols were selected and validated by the optimized method in three wine matrices. The overall analytical performances demonstrated by this method were equivalent, and in most cases, greater than many previously developed GC–MS or LC–MS methods. The validated method was applied to analyze a selection of wines in which 12 thiols were quantified.

Identifying biomarkers for nasopharyngeal carcinoma diagnosis and chemoradiotherapy response using serum endogenous small molecules profiling

Changes in metabolic characteristics are important features of tumor progression and prognosis, including nasopharyngeal carcinoma (NPC). Identifying serum metabolites as potential diagnostic and chemoradiotherapy response biomarkers for NPC is therefore crucial. In this study, ultra-performance liquid chromatography coupled with linear ion trap quadrupole orbitrap high-resolution mass spectrometry (UPLC-LTQ-Orbitrap MS) was used to analyze metabolic variations among controls, NPC patients, and NPC patients undergoing chemoradiotherapy (CRT). Univariate and multivariate analyses revealed seven differential metabolites between the control and NPC groups and eleven metabolites between the CRT and NPC groups. Five common metabolites, gluconic acid, palmitic acid, LysoPC (15:0/0:0), stearic acid, and LysoPC (20:2(11Z,14Z)/0:0), were consistently altered across groups. Notably, the first four metabolites were adjusted closer to normal after chemoradiotherapy, while this change is not reflected at LysoPC (20:2(11Z,14Z)/0:0). These common metabolites were enriched in five pathways. These findings underscore the importance of serum metabolite profiling in NPC diagnosis and treatment response assessment and offer a promising foundation for further clinical research.

Optimisation and validation of liquid-liquid extraction of aminoglycosides from foods of animal origin

ABSTRACT This study focused on evaluating and optimising the liquid-liquid extraction (LLE) method for the multi-residue analysis of seven AGs in bovine liver, meat, milk, and kidney in conjunction with liquid chromatography coupled to a quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS) for quantification. Of all the hydrophilic interaction liquid chromatography (HILIC) stationary phases tested, the ZIC-HILIC column yielded the best performance. The method was validated according to Commission Implementing Regulation (CIR) EU 2021/808, which encompassed various concentration ranges from 25–7500 µg/kg. The mean recoveries ranged from 79% for liver matrix to 87% for meat matrix, with coefficients of variation (CV) below 18%. The method exhibited excellent linearity, as indicated by correlation coefficients (R2) greater than 0.9903 across all tested matrices. The decision limit (CCα) and detection capability (CCβ) varied from 62 to 5984 µg/kg and 71 to 6471 µg/kg, respectively. The limits of detection (LOD) and quantification (LOQ) were established between 1 and 45 µg/kg, and 2 and 75 µg/kg, respectively. No AG residues were detected in the 120 samples that were collected from different local markets in Egypt using the developed analytical method.

Rapid Determination of 12 Classes of Per- and Polyfluoroalkyl Substances in Water Samples from Environmental Forensic Cases.

The widespread use of per- and polyfluoroalkyl substances (PFASs) with different physico-chemical properties poses a great threat to the environment and human health. Simultaneous detection of different classes of PFASs is a difficult task, especially for rapid analysis of polluted water samples in environmental forensic cases. In this study, a simple sample preparation ultrahigh-performance liquid chromatography coupled with quadrupole Orbitrap high-resolution mass spectrometry was established for the detection of PFASs in a wide range of water matrices. By optimizing the conditions of pretreatment and the parameters of the instrument, the developed method provided good linearity of calibration standards (R2 > 0.99), and demonstrated excellent MLOQ (0.008-1.2 µg/L), with spiked recoveries ranging from 57.7% to 151% for 47 targets in surface water samples, and from 45.7 to 165% for 46 targets in ground and waste water samples, respectively. This method required an injection volume of 3 µL and an analysis time of only 18 min per sample. The validation method was successfully applied to the analysis of 20 environmental water samples, in which 15 target substances with different concentrations were detected, with total concentrations of 0.082 to 262.455 μg/L. The method is simple and exclusive, and can rapidly confirm the occurrence of PFASs in different water samples, providing a convenient and fast high-throughput analysis, which is especially suitable for the application in the environmental forensic investigation of PFASs pollution.

Suspect screening of per-and polyfluoroalkyl substances in paper by selective and non-selective extraction with UHPLC-Q orbitrap MS

Non-targeted analysis and suspect screening of per- and polyfluoroalkyl substances (PFAS) in various matrices have gained traction with advancements in accurate mass analytical instruments. This study employed ultra-high performance liquid chromatography coupled to quadrupole orbitrap high-resolution mass spectrometry for PFAS suspect screening of paper grades used in the paper recycling chain. The samples were prepared using two extraction techniques; selective accelerated solvent extraction with weak anionic exchange solid-phase extraction and non-selective ultrasonic-assisted extraction. A suspect screening protocol was established to tentatively identify suspected PFAS against spectral databases using a systematic approach of peak filtering and study-specific thresholds for reporting, linked to a confidence level. The possible prevalence of previously unreported PFAS in several paper materials across the various collection sites in the paper recycling chain was inferred by the common detection of short-chain polyfluoroalkyl ketones and diketones in the paper recycling chain. The suspect screening tentatively identified 41 unique PFAS, with 3 common to both pre-treatment techniques. The detection of unique PFAS by the two sample pre-treatment techniques highlighted the significance of both selective and non-selective extraction in PFAS screening endeavours. Further, it showed the importance of understanding the acquisition mechanisms employed in mass spectrometry where data-dependent acquisition triggered fragmentation in certain identified compounds, and not in others. The tentatively identified PFAS indicated that there were several previously unreported PFAS in the paper recycling chain and that additional studies were required to investigate their abundance, possible persistence, bioaccumulation and toxicity, in relation to their functional groups and carbon chains.

Degradation of Isotianil in Water and Soil: Kinetics, Degradation Pathways, Mechanisms, and Ecotoxicity Assessments.

Pesticides are transported and transformed in soil and can enter surface water through various pathways. They undergo hydrolysis, oxidation, and photoconversion in surface water. Isotianil is a new fungicide that effectively controls rice blast. However, there are limited reports on its degradation. Herein, the hydrolysis and photolysis of isotianil in water and its degradation in soil samples from five provinces of China were investigated. The degradation products of isotianil were identified using ultrahigh-performance liquid chromatography-Q exactive hybrid quadrupole-orbitrap high-resolution accurate mass spectrometry, and four compounds were discovered for the first time. The degradation pathways of isotianil were inferred, and the reaction active site and degradation mechanism of isotianil were clarified based on density functional theory calculations. The ecotoxicity of the degradation product M118 (aminobenzonitrile) was found to be moderate toward Daphnia magna, which was predicted and confirmed by Ecological Structure Activity Relationships and the experiment, respectively. The results of this study will contribute to a better understanding of the fate of isotianil in the environment.

Quick and high-throughput quantification of β-agonist residues in bovine liver, meat, milk, kidney, poultry, and egg using dispersive solid phase extraction

A reliable liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS) method was developed for the simultaneous identification and quantification of 13 β-agonist residues in bovine liver, meat, milk, kidney, poultry, and egg. Dispersive-solid phase extraction (d-SPE) using acetonitrile (ACN) was used to prepare the samples. The analyte in the extracts was separated on a reversed-phase Accucore aQ (50 mm × 2.1 mm, 2.6 μm) using a mobile phase of an aqueous solution containing 2 mM ammonium acetate and acetonitrile (ACN) 0.1 % formic acid. The method was validated in accordance with Commission Implementing Regulation (CIR) EU 2021/808 at six different concentrations ranging from 0.1 to 5 μg/kg. The mean recoveries ranged from 65 to 94 %, while repeatability and reproducibility values were all below 13 %. The linearity, as correlation coefficients (R2) ranged from 0.9955 to 0.9999. The decision limit (CCα) and detection capability (CCβ) ranges were 0.11–0.13 µg/kg and 0.12–0.15 µg/kg, respectively. The limits of detection (LOD) and limits of quantification (LOQ) were in the range of 0.004–0.048 μg/kg and 0.010–0.075 μg/kg, respectively. Of the 180 samples that were collected from local markets in Egypt, 21.11 % had β-agonist residues. The mean concentration (µg/kg) and detection frequency (%) of the most frequently found β-agonist in the samples were as follows: terbutaline (2.63 µg/kg and 90 %), ractopamine (5.14 µg/kg and 23.3 %). The method's applicability was verified by successfully completing two rounds of proficiency testing (PT).

Exploring the gut microbiota mediated biotransformation of Senecio scandens Buch.-Ham.: Insights from metabolite spectrum with UHPLC-Q-Orbitrap HRMS and bioinformatics analysis of gut microbiota metabolites

Senecio scandens Buch.-Ham., a traditional Chinese medicine commonly used clinically, exhibits various pharmacological properties, including anti-inflammatory, anti-tumor, antiviral, and antibacterial activities. However, its water extracts' chemical components and metabolites are inadequately understood, limiting further research. In this study, the chemical components and metabolism processes of Senecio scandens, both in vivo (plasma, feces, urine, and bile) and in vitro (gut microbiota and liver microsomes), were characterized based on ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry. Additionally, metabolites detectable in fecal samples and intestinal microbiota incubated but absent in liver microsomes were identified as characteristic metabolites of intestinal microbiota. The targets of the characteristic metabolites of intestinal microbiota were collected, followed by exploration of potential pathways through KEGG enrichment analysis. As a result, a total of 133 chemical components were preliminarily identified, including 35 organic acids, 21 alkaloids, 19 flavonoids and their glycosides, 17 phenylpropanoids, 10 jacaranda ketones, and 31 other compounds. Notably, 12 of these were potentially novel compounds. In addition, 39 prototype components in rats and 109 metabolites were identified and characterized, including 102 in vivo and 52 metabolites in vitro (51 in rat gut microbiota and 24 in rat liver microsomes). The main metabolic pathways include oxidation, reduction, hydrolysis, methylation, glucuronidation, sulfonation, and acetylation reactions. Furthermore, KEGG enrichment analysis revealed that the characteristic metabolites of intestinal microbiota may be related to the ErbB, FoxO, mTOR, and MAPK signaling pathways, exhibiting anti-inflammatory and anti-tumor effects. In summary, the chemical components and metabolites of Senecio scandens were comprehensively identified using a rapid and accurate method, providing a scientific basis for the in-depth study of the material basis and its clinical application of Senecio scandens.

Multiclass method for detecting 41 antibiotic residues in bovine liver, muscle, and milk using LC-Q-Orbitrap-HRMS

This study developed and validated an analytical method for the simultaneous identification and quantification of 41 veterinary antibiotic drugs from 9 different classes, including sulfonamides, quinolones, tetracyclines, penicillins, macrolides, nitroimidazoles, cephalosporins, diaminopyrimidines, and amphenicol, in animal-derived food products. The method used modified QuEChERS and liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS). The method was validated in accordance with Commission Implementing Regulation (CIR) EU 2021/808 at five different concentrations ranging from 0.075 to 10,000 μg kg−1. The mean recoveries ranged from 58 % to 123 %, while repeatability values were all below 21 %. The calibration curves showed good linearity, with correlation coefficients (R2) ranging from 0.9905 to 0.9999. The limits of detection (LOD) and limits of quantification (LOQ) were in the range of 0.005–43.0 μg kg−1 and 0.008–71.7 μg kg−1, respectively. The decision limit (CCα) and detection capability (CCβ) ranges were 0.37–2347 µg/kg and 0.44–3099 µg/kg, respectively. Out of the 1330 samples collected from Egyptian local markets, 1.20 % had antibiotic residues. The applicability of the developed method was proven through successful four proficiency testing (PT). The proposed method was demonstrated to be reliable for the simultaneous analysis of multiclass veterinary drugs in bovine liver, muscle tissue, and milk.

Optimization and validation of metabolomics methods for feline urine and serum towards application in veterinary medicine

BackgroundMetabolomics is an emerging and powerful technology that offers a comprehensive view of an organism's physiological status. Although widely applied in human medicine, it is only recently making its introduction in veterinary medicine. As a result, validated metabolomics protocols in feline medicine are lacking at the moment. Since biological interpretation of metabolomics data can be misled by the extraction method used, species and matrix-specific optimized and validated metabolomic protocols are sorely needed. ResultsSystematic optimization was performed using fractional factorial experiments for both serum (n = 57) and urine (n = 24), evaluating dilution for both matrices, and aliquot and solvent volume, protein precipitation time and temperature for serum. For the targeted (n = 76) and untargeted (n = 1949) validation of serum respectively, excellent instrumental, intra-assay and inter-day precision were observed (CV ≤ 15% or 30%, respectively). Linearity deemed sufficient both targeted and untargeted (R2 ≥ 0.99 or 0.90, respectively). An appropriate targeted recovery between 70 and 130% was achieved. For the targeted (n = 69) and untargeted (n = 2348) validation of the urinary protocol, excellent instrumental and intra-assay precision were obtained (CV ≤ 15% or 30%, respectively). Subsequently, the discriminative ability of our metabolomics methods was confirmed for feline chronic kidney disease (CKD) by univariate statistics (n = 41 significant metabolites for serum, and n = 55 for urine, p-value<0.05) and validated OPLS-DA models (R2(Y) > 0.95, Q2(Y) > 0.65, p-value<0.001 for both matrices). SignificanceThis study is the first to present an optimized and validated wholistic metabolomics methods for feline serum and urine using ultra-high performance liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry. This robust methodology opens avenues for biomarker panel selection and a deeper understanding of feline CKD pathophysiology and other feline applications.

Anti-aging mechanism and effect of treatment with raw and wine-steamed Polygonatum sibiricum on D-galactose-induced aging in mice by inhibiting oxidative stress and modulating gut microbiota.

Polygonatum sibiricum (PS) is a traditional Chinese medicine (TCM) first recorded in Mingyi Bielu. The book documents that PS can nourish five internal organs, be taken for a long time, relax the body and prolong lifespan. Presently, PS is widely used in TCM to prevent premature graying of hair. Based on TCM theory and clinical trials, the wine steaming processed product from PS provides a better effect. However, no published study has elucidated the anti-aging mechanism. The study aim was to investigate the anti-aging mechanism of PS and its wine steaming processed product in mice, specifically focusing on the effect of D-galactose (D-gal) surrounding the intestinal flora and the Kelch-like ECH-associated protein 1-nuclear factor erythroid 2-related factor 2-antioxidant response elements (Keap1/Nrf2/ARE) pathway. The chemical components in Raw PS (RPS) and Wine-steamed PS (WPS) were identified by ultra-performance liquid chromatography-hybrid quadrupole-Orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap HRMS). An aging model using Kunming mice was established through intraperitoneally injected D-gal. Concentrations of RPS and WPS at 5, 10, or 15g/kg/day levels were administered intragastrically, respectively. The body weight, liver and spleen indexes, superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), and malondialdehyde (MDA) activities in serum and brain tissue were recorded. Hematoxylin and eosin (HE) stained brain tissue was histopathologically examined. The expressions of Keap1, Nrf2 and heme oxygenase 1 (HO-1) in the brain tissue at the mRNA and protein levels were respectively detected by reverse transcription-polymerase chain reaction (RT-PCR) and western blot (WB). Moreover, an Illumina Hiseq platform was used for 16S ribosomal RNA (16S rRNA) high-throughput sequencing to evaluate the proportions of intestinal flora in aging mice. The proportions of saccharides, flavonoids, and triterpene acids were different between RPS and WPS. In the aging model mice, WPS outperformed RPS in improving body weight and mental state by increasing the spleen index, SOD and GSH-PX activities, decreasing the liver index and MDA activities, and restoring the histopathological morphology in D-gal-induced aging mice. At the mRNA levels, RPS and WPS significantly reduced the expression of Keap1 and increased the expressions of Nrf2 and HO-1. The trend in protein expressions was similar to that of the mRNA results, and WPS had a stronger effect than RPS. Fecal microbiota analysis showed that RPS and WPS restored intestinal microbiota proportions to normal levels. The results demonstrated that PS and its WPS had a positive effect in relieving oxidative stress in aging mice. WPS outperformed RPS, which might be related to the activation of the Keap1/Nrf2/ARE pathway and regulation of intestinal flora.

Untargeted Metabolomics Based on Ultra-High-Performance Liquid Chromatography Coupled with Quadrupole Orbitrap High-Resolution Mass Spectrometry for Differential Metabolite Analysis of Pinelliae Rhizoma and Its Adulterants.

The present study investigates the chemical composition variances among Pinelliae Rhizoma, a widely used Chinese herbal medicine, and its common adulterants including Typhonium flagelliforme, Arisaema erubescens, and Pinellia pedatisecta. Utilizing the non-targeted metabolomics technique of employing UHPLC-Q-Orbitrap HRMS, this research aims to comprehensively delineate the metabolic profiles of Pinelliae Rhizoma and its adulterants. Multivariate statistical methods including PCA and OPLS-DA are employed for the identification of differential metabolites. Volcano plot analysis is utilized to discern upregulated and downregulated compounds. KEGG pathway analysis is conducted to elucidate the differences in metabolic pathways associated with these compounds, and significant pathway enrichment analysis is performed. A total of 769 compounds are identified through metabolomics analysis, with alkaloids being predominant, followed by lipids and lipid molecules. Significant differential metabolites were screened out based on VIP > 1 and p-value < 0.05 criteria, followed by KEGG enrichment analysis of these differential metabolites. Differential metabolites between Pinelliae Rhizoma and Typhonium flagelliforme, as well as between Pinelliae Rhizoma and Pinellia pedatisecta, are significantly enriched in the biosynthesis of amino acids and protein digestion and absorption pathways. Differential metabolites between Pinelliae Rhizoma and Arisaema erubescens are mainly enriched in tyrosine metabolism and phenylalanine metabolism pathways. These findings aim to provide valuable data support and theoretical references for further research on the pharmacological substances, resource development and utilization, and quality control of Pinelliae Rhizoma.

Development and validation of a multiclass method for the determination of veterinary drug residues in honey

ABSTRACT This study developed and validated an analytical method for simultaneous identification and quantification of 41 veterinary drugs in honey using liquid liquid extraction (LLE) and liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap HRMS). The method was validated in accordance with Commission Implementing Regulation (CIR) EU 2021/808 at five different concentrations ranging from 0.075 to 50 μg/kg. The mean recoveries ranged from 70 to 105, while repeatability values were all below 17%. The linearity, as correlation coefficients (R2) ranged from 0.994 to 1. The limits of detection (LOD) and limits of quantification (LOQ) were in the range of 0.006–3.92 μg/kg and 0.011–6.54 μg/kg, respectively. The decision limit (CCα) and detection capability (CCβ) ranges were 0.0759–5.33 µg/kg and 0.0804–6.08 µg/kg, respectively. Of the 263 honey samples that were collected from local markets in Egypt, 47.5% had antibiotic residues. The mean concentration (µg/kg) and detection frequency (%) of the five most frequently detected antibiotics in the honey samples were as follows: trimethoprim (143.25 µg/kg and 39.9%), sulfamethoxazole (136.69 µg/kg and 30.7%), sulphadiazine (77.19 µg/kg and 18.6%), tylosin (184.37 µg/kg and 18.2%), and ciprofloxacin (185.33 µg/kg and 7.60%). The applicability of the developed method was proven through successful three proficiency testing (PT). The proposed method was demonstrated to be reliable for the simultaneous analysis of multi-class veterinary drugs in honey.

Exploring anti-inflammatory and antioxidant-related quality markers of Artemisia absinthium L. based on spectrum-effect relationship.

Artemisia absinthium L. is a well-known medicinal, aromatic, and edible plant with important medicinal and economic properties and a long history of use in treating liver inflammation and other diseases; however, there has been insufficient progress in quality control. This study aimed to investigate the quality markers for the anti-inflammatory and antioxidant activities of A.absinthium based on spectrum-effect relationship analysis. Eighteen batches of A.absinthium from different origins were used. Chemical fingerprints were obtained by ultra-performance liquid chromatography (UPLC). The chemical compositions were identified by quadrupole-Orbitrap high-resolution mass spectrometry. Anti-inflammatory activity was assessed by inhibition of cyclooxygenase-2 and 15-lipoxygenase in vitro and inhibition of nitric oxide release in lipopolysaccharide-induced BV-2 cells. Antioxidant activity was assessed by DPPH and ABTS radical scavenging assays. The relationship between bioactivity and chemical fingerprints was then analyzed using chemometrics including gray relational analysis, bivariate correlation analysis, and orthogonal partial least squares analysis. Different batches of A.absinthium extracts possessed significant anti-inflammatory and antioxidant activities to varying degrees. Eighty compounds were identified from A.absinthium, and 12 main common peaks were obtained from the UPLC fingerprints. P3 (chlorogenic acid), P5 (isochlorogenic acid A), and P6 (isochlorogenic acid C) were screened as the most promising active compounds by correlation analysis and further validated for their remarkable anti-inflammatory effects. This is the first study to screen the quality markers of A.absinthium by establishing the spectrum-effect relationship, which can provide a reference for the development of quality standards and further research on A.absinthium.

Degradation of co-formulants and metabolites from plant protection products in courgette and tangerine using LC-HRMS: Laboratory tests

Plant protection products contain co-formulants that could end up in vegetables, and they can generate transformation products that can be more toxic than their original molecule, which are a potential risk to food safety. Therefore, this study evaluated the dissipation of two co-formulants, dodecylbenzenesulfonic acid and 1-ethyl-2-pyrrolidone, in courgette and tangerine samples to determine possible harmful metabolites under laboratory conditions. The analyses of these substances were performed by liquid chromatography coupled to quadrupole-Orbitrap high-resolution mass spectrometry (LC-Q-Orbitrap-HRMS). For the degradation of both compounds, a single-phase kinetic model was fitted, with R2 values greater than 0.99. In courgette and tangerine, half-lives (DT50) for dodecylbenzene sulfonic acid were 1.83 and 1.42 days, while DT50 was 6.26 and 5.04 days for 1-ethyl-2-pyrrolidone, respectively. Three metabolites of dodecylbenzenesulfonic acid and five metabolites of 1-ethyl-2-pyrrolidone were found in courgette, while the same metabolites were detected in tangerine samples, except benzoic acid and 4-aminobutanoic acid. These metabolites were identified for the first time from these compounds except for benzoic acid, observing that 5-hydroxy-N-ethylpyrrolidone was the most concentrated metabolite in tangerine 14 days after application, reaching a maximum concentration of 149 µg/kg. Furthermore, some of the detected metabolites possessed a value of LD50 lower than their original molecules. In consequence, such metabolites, derived from these co-formulants, should be controlled to prevent negative health effects, and ensure food safety.

Systematic qualitative analysis of terpenes in mastic (Pistacia lentiscus L.) extract and their fragmentations by UHPLC-Q-Orbitrap-HRMS.

Mastic is a natural resin produced by Pistacia lentiscus L. (Anacardiaceae). The beneficial properties of this resin are attributed to its triterpenes and volatile compounds. This study was conducted to screen and characterize the terpenes in mastic ethyl acetate extract (M-Ex). An ultrahigh-performance liquid chromatography coupled to quadrupole Orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) method was developed for the qualitative analysis of terpenes in M-Ex. We utilized in-house-isolated compounds as reference substance (Rs), including monoterpenes (A) with α-pinane structures, tetracyclic triterpene (B) containing tirucallane skeletons, and pentacyclic triterpene (C) belonging to olean, moronic, amyrone, and lupane types. Based on the mass spectrometric characteristics of the above compounds, and the difference in characteristic diagnostic fragment ions (DFIs) in isomeric compounds, the terpene compounds were further identified in M-Ex. Out of a total of 70 compounds, including monoterpenes and tetra-, and pentacyclic triterpenes, 20 were accurately determined by Rs, retention time (RT), and DFIs. Based on the cleavage patterns summarized from the above 20 compounds and with reference to the reported literature, another 50 compounds were putatively identified. Based on our discovery, six terpenic acids with A-seco-tirucallane types and one monoterpene dimer were identified for the first time in mastic. Our research serves not only as a foundation for the rapid identification and screening of terpene compounds in mastic but also as a supplementary basis for the identification of such compounds in other types of resins.

Quality assessment and Q-markers discovery in Citri Sarcodactylis Fructus by integrating serum pharmacochemistry and network pharmacology.

Citri Sarcodactylis Fructus (CSF), a common fruit and traditional Chinese medicine (TCM), has been hindered in its further development and research owing to the lack of comprehensive and specific quality evaluation standards. This study aimed to establish clear TCM quality standards related to the therapeutic mechanisms of CSF and to provide a basis for subsequent research and development. Ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-orbitrap HRMS) technology was used to comprehensively identify CSF components and explore their absorbance levels in rat serum. Network pharmacology research methods were employed to investigate the potential mechanisms of action of the identified components in the treatment of major clinical diseases. Subsequently, a combination of HPLC chromatographic fingerprinting for qualitative analysis and multi-index content determination was used to evaluate the detectability of the identified quality markers (Q-markers). Twenty-six prototype components were tentatively characterized in rat serum. Network pharmacology analysis showed six effective components, namely 7-hydroxycoumarin, isoscopoletin, diosmin, hesperidin, 5,7-dimethoxycoumarin, and bergapten, which played important roles in the treatment of chronic gastritis, functional dyspepsia, peptic ulcer, and depression and were preliminarily identified as Q-markers. The results of content determination in 15 batches of CSF indicated significant differences in the content of medicinal materials from different origins. However, compared with the preliminarily determined Q-markers, all six components could be measured and were determined as Q-markers of CSF. The chemical Q-markers obtained in this study could be used for effective quality control of CSF.