High-performance Liquid Chromatography-mass Spectrometry Research Articles

In vitro simulated digestion and Caco-2 cell absorption to explore potential of soy protein isolates as whey protein substitutes in canine and feline food.

Digestive properties and bioavailability of soy protein isolate and whey protein in canine and feline were evaluated using in vitro simulated digestion and cell absorption experiments. The amino acids and peptides in soybean protein isolate and whey protein after digestion were detected by high-performance liquid chromatography and liquid mass spectrometry. Gastrointestinal digestion rates of soy protein isolate and whey protein were analyzed by fitting digestive kinetics curves. The degree of absorption and utilization of amino acids and peptides from the digestive products was evaluated by establishing a Caco-2 cell model. The results demonstrated that soy protein isolate, like whey protein, could produce large amounts of free amino acids and peptides distributed 100-1500 Da. The gastrointestinal digestion speed of soy protein isolate was similar with whey protein. It is worth noting from the intestinal absorption of Caco-2 cell that more than 15 amino acids produced by soy protein isolate and whey protein could be absorbed, and a large number of peptides were also utilized by Caco-2 cell. The bioavailability of soy protein isolates and whey protein for dogs respectively reached 6.30% and 9.00%, and it reached 7.40% and 16.40% in cats, respectively. Soy isolate proteins can be digested and absorbed by pets like whey proteins. These findings may provide significant strategies and support for the application of plant-based proteins in pet foods.

Effects of fermented tea by Aspergillus cristatus on the characteristic aroma and non-volatile components of Jiang-flavor baijiu.

Fu Brick tea (FBT) extract has been demonstrated to lower blood lipids, protect liver, and prevent obesity. Despite these benefits, there are no products on the market that combine FBT with other foods or beverages. In this study, we developed a novel product by combining FBT with high-alcohol Jiang-flavor baijiu, resulting in FBT-baijiu. High-performance liquid chromatography-mass spectrometry (HPLC-MS) analysis revealed that FBT-baijiu contains health-promoting alkaloids, including eurocristatine (13.60 ± 0.13 mg/L), (-)-neoechinulin A (10.26 ± 0.09 mg/L), neoechinulin D (7.89 ± 0.05 mg/L), variecolorin G (6.94 ± 0.05 mg/L), and echinulin (25.46 ± 0.27 mg/L), which are known to be present in FBT. The aroma compounds of the FBT-baijiu and the base baijiu were analyzed using comprehensive gas chromatography-olfactometry-mass spectrometry (GC-O-MS) technology. The major volatile compounds of two baijiu samples were identified using relative odor activity values (r-OAVs) analysis. The FBT-baijiu showed a significant reduction in ester content, but a remarkable enhancement in aromatic properties. The findings show that combining FBT with baijiu not only offers functional components but also enhances the flavor profile. This study highlights the potential of utilizing FBT as a functional food ingredient or additive, paving the way for the development of new health-promoting products. © 2024 Society of Chemical Industry.

Spatial Variations in Per- and Polyfluoroalkyl Substance Concentrations in Pooled Sera from Inland, Coastal, and Island Populations

Per- and polyfluoroalkyl substances (PFAS) are a class of ubiquitously detected chemicals, some of which are highly persistent and bioaccumulative in humans. Within the general population, dietary ingestion is considered a primary pathway for PFAS exposure, and seafood consumption specifically has been associated with higher serum PFAS concentrations. Proximity of residence to the ocean may influence dietary habits, particularly seafood consumption, and exposure to geographically specific PFAS sources such as sea spray aerosols (SSA). The objective of this study was to evaluate potential spatial trends in serum PFAS concentrations between Australian coastal and island populations compared to those with inland residency. Human sera were obtained from deidentified surplus pathology samples and pooled with respect to geographical location, sex (male or female), and age group (males: ≥15-<45 years, ≥45 years; females: ≥15-<45 years, ≥45-<60 years, ≥60 years) stratification criteria. Serum samples were then analysed for PFAS using High Performance Liquid Chromatography-Mass Spectrometry (HP LC-MS). A total of 13 of the 45 targeted PFAS were quantifiable in at least one pooled sample, including the detection of perfluorooctane sulfonate (PFOS) replacement compounds 5:3 fluorotelomer carboxylic acid (5:3 FTCA) and potassium 9-chlorohexadecafluoro-3-oxanonane-1-sulfonate (9Cl-F53B). Significant spatial trends were observed in males aged ≥45 years, with serum concentrations of PFOS, perfluorobutanoic acid (PFBA), perfluorodecanoic acid (PFDA) and perfluoroheptane sulfonic acid (PFHpS) demonstrated to be 32-77% higher in pooled samples from island locations compared with inland. A similar trend was observed for PFHpS in coastal locations. Whilst deidentification of samples limited inferences about exposure pathways associated with the observed trends, this study indicated the feasibility of utilising pooled samples for assessing spatial variations in serum PFAS concentrations between geographically distinct subpopulations.

Transforming growth factor-β3/Smad2/Smad3 signaling pathway inhibition and autophagy by the Yunpi-Xiefei-Huatan decoction ameliorated airway inflammation and mucus hypersecretion in asthmatic rats

Ethnopharmacological relevanceThe Yunpi-Xiefei-Huatan decoction (YXHD) is a traditional Chinese medicine that can improve asthma-related symptoms, including cough, phlegm in the throat, and shortness of breath. However, the YXHD mechanism on asthma has not yet been elucidated. Study aimThe aim of this study is to investigate the effect of YXHD on airway inflammation, mucus hypersecretion, and autophagy modulation in asthma. Materials and methodsThe YXHD chemical constituents were observed and analyzed using high-performance liquid chromatography-mass spectrometry. Ovalbumin sensitization and stimulation were used to establish an asthma rat model. A total of 80 Sprague-Dawley (SD) rats were segmented into eight groups at random: a Normal (NC) group, a Model (Mod) group, a YXHD low-dose group (10 g/kg/d), a YXHD moderate-dose group (20 g/kg/d), a YXHD high-dose group (40 g/kg/d), a Rapamycin group (4 mg/kg/d), a 3-methyladenine (3-MA) group (15 mg/kg/d), and a Dexamethasone (DEX) group (0.5 mg/kg/d). Whole-body plethysmography (WBP) detection was used to evaluate airway hyperresponsiveness. An enzyme-linked immunosorbent assay (ELISA) was used to detect inflammatory factors in the peripheral blood. Inflammatory cells in the bronchoalveolar lavage fluid (BALF) were also counted. Pathological changes in the lung tissues were marked using hematoxylin and eosin (H&E) staining and periodic acid-Schiff (PAS) staining. The localization of MUC5AC and the co-localization of LC3B + MUC5AC were observed using immunofluorescence. The expressions of autophagy and the TGF-β3/Smad2/Smad3 pathway in the lung tissues were detected using a Western blot assay (WB) and qPCR, and the autophagosomes in the lung tissues were detected using a transmission electron microscope (TEM). ResultsTwenty signal peaks were identified using ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) technology. The TGF-β3/Smad2/Smad3 signal pathway activation was induced using ovalbumin (OVA) exposure in the rats. The upregulated expression of autophagy, enhanced MUC5AC fluorescence and LC3B fluorescence, and their co-localized expression in the airway epithelium indicated inflammatory cell infiltration and excessive mucus secretion in the lungs. This resulted in airway hyper-responsiveness. The YXHD inhibited the activation of the TGF-β3/Smad2/Smad3 signaling pathway, and autophagy expression reduced inflammatory factors, abnormal mucus secretion, and airway hyperresponsiveness. ConclusionThe YXHD improved lung function, relieved lung inflammation, and inhibited airway mucus secretions in asthmatic rat models. Its mechanism may have been related to the blockage of the TGF-β3/Smad2/Smad3 signaling pathway and autophagy downregulation.

Relationship between metabolomics of T2DM patients and the anti-diabetic effects of Phellodendri Chinensis Cortex-Anemarrhenae Rhizoma herb pair in mice

Ethnopharmacological relevanceType 2 diabetes mellitus (T2DM) poses significant threats to public health. In Traditional Chinese Medicine (TCM), the Phellodendri Chinensis Cortex-Anemarrhenae Rhizoma (PCC/AR) herb pair has long been used for T2DM treatment, although its specific anti-diabetic mechanisms remain unclear. Aim of the studyThis study aimed to elucidate the relationship between metabolomics of T2DM patients and the anti-diabetic effects of PCC/AR herb pair in mice through clinical metabolomics and both in vitro and in vivo experiments. Materials and methodsIn this study, a T2DM mouse model was established via high-fat feeding (HFD) and streptozotocin (STZ) injection. The effects of PCC/AR on blood glucose, lipid metabolism, and inflammatory markers were evaluated. High-performance liquid chromatography-mass spectrometry (HPLC-MS) was performed for metabolomics analysis of T2DM patients. ResultsSerum metabolomics analysis identified significant alterations in metabolites linked to the biosynthesis of unsaturated fatty acids and purine metabolism in T2DM patients, with elevated 2-hydroxyvaleric acid (2HB) levels. In T2DM mice, PCC/AR intervention normalized FBG, GHbA1c, TC, TG, LDL-C, HDL-C, TNF-α and IL-1β levels, while improving insulin sensitivity and pancreatic β-cell function in T2DM mice. Notably, PCC/AR reduced key enzymes in gluconeogenesis and fatty acid synthesis, PEPCK and ACC1. ConclusionPCC/AR herb pair exerts an anti-diabetes effect in T2DM mice by regulating 2HB through ACC1 inhibition, thereby reducing FFA and TG synthesis. Additionally, PCC/AR may also exert its effects by modulating glucose and lipid metabolism and reducing inflammation. These results support further investigation into the PCC/AR herb pair as a complementary therapy for T2DM.

Proximal characteristics, phenolic compounds profile, and functional properties of Ullucus tuberosus and Arracacia xanthorrhiza

Aim: The Andean tubers Ullucus tuberosus and Arracacia xanthorrhiza are of great international importance due to their nutritional value (carbohydrates, fiber), functional and phenolic compounds. This study aimed to determine the proximate composition of the flours of these tubers, their functional properties, and the phenolic profiles of both the skin and the pulp, since the flour used is a mixture of both. Methods: The proximal characteristics were determined as established by the Association of Official Analytical Chemists (AOAC), the functional properties, water holding capacity (WHC), swelling capacity (SC), and oil holding capacity (OHC); and the profile of phenolic compounds was carried out by high performance liquid chromatography-mass spectrometry (HPLC-MS). Results: Results revealed that the tubers contain more than 50% carbohydrates, with U. tuberosus having 14% fiber and A. xanthorrhiza having 4.07% fiber. Notably, they are rich in minerals, such as potassium (K), with levels of 4.12% and 3.32% for U. tuberosus and A. xanthorrhiza, respectively. In terms of functional properties, U. tuberosus exhibited a WHC of 0.78 g water/g dry matter (DM), an SC of 1.42 g water/g DM, and an OHC of 0.44 g oil/g DM. In contrast, A. xanthorrhiza showed a WHC of 0.96 g water/g DM, an SC of 3.86 g water/g DM, and an OHC of 0.18 g oil/g DM. Additionally, the study identified various hydroxycinnamic acids, including caffeic acid 4-O-glucoside and p-coumaroyl glucose, as well as methoxyflavonols, such as 3,7-dimethylquercetin. Conclusions: These characteristics suggest that the flours could be valuable for the production of bakery products, pastries, pastas, or foods requiring high viscosity. Furthermore, U. tuberosus and A. xanthorrhiza flours have potential applications in the development of functional foods, thus promoting their use and adding value to these tubers produced in the Andean region.

In utero exposure to per - and polyfluoroalkyl substances (PFAS) associates with altered human infant T helper cell development.

Environmental exposures to chemical toxicants during gestation and infancy can dysregulate multiple developmental processes, causing lifelong effects. There is compelling evidence of PFAS-associated immunotoxicity in adults and children. However, the effect of developmental PFAS exposure on infant T-cell immunity is unreported, and, if present, could be implicated in immune-related health outcomes. We seek to model longitudinal changes in CD4+ T-cell subpopulations from birth through 12 months and their association with in-utero PFAS exposure and postnatal CD4+ T-cell frequencies and functions. Maternal-infant dyads were recruited as part of the UPSIDE-ECHO cohort during the first trimester between 2015 and 2019 in Rochester, New York; dyads were followed through the infant's first birthday. Maternal PFAS concentrations (PFOS, PFOA, PFNA, and PFHXS) were quantified in serum during the second trimester using high-performance liquid chromatography and tandem mass spectrometry. Infant lymphocyte frequencies were assessed at birth, 6- and 12-months using mass cytometry and high-dimensional clustering methods. Linear mixed-effects models were employed to analyze the relationship between maternal PFAS concentrations and CD4+ T-cell subpopulations (n=200). All models included a PFAS and age interaction and were adjusted for parity, infant sex, and pre-pregnancy body mass index. In-utero PFAS exposure correlated with multiple CD4+ T-cell subpopulations in infants. The greatest effect sizes were seen in T-follicular helper (Tfh) and T-helper 2 (Th2) cells at 12 months. A log 2 -unit increase in PFOS was associated with lower Tfh [0.17% (95%CI: -0.30, -0.40)] and greater Th2 [0.27% (95%CI: 0.18, 0.35)] cell percentages at 12 months. Similar trends were observed for PFOA, PFNA, and PFHXS. Maternal PFAS exposures correlate with cell-specific changes in the infant T-cell compartment, including key CD4+ T-cell subpopulations that play central roles in coordinating well-regulated, protective immunity. Future studies into the role of PFAS-associated T-cell distribution and risk of adverse immune-related health outcomes in children are warranted.

Manipulating flavonoid biosynthesis in Trigonella persica through controlled spectral lighting

Trigonella persica Boiss., is renowned for its rich phytochemical profile, particularly the presence of the flavonol quercetin. This study explored the effects of various light treatments, including blue, red, blue-red (1:1) radiation (BRR), and pink fluorescent light (PFL), on the biochemical and molecular mechanisms governing quercetin and flavonoid biosynthesis in T. persica at different growth stages. Our results showed that light treatments significantly influenced the activity of key enzymes phenylalanine ammonia-lyase (PAL) and tyrosine ammonia-lyase (TAL) during germination and vegetative growth, with blue light inducing higher PAL and TAL activities compared to control conditions. High-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS) analyses revealed that 48-h-old sprouts grown under red light exhibited the highest levels of flavonoid components and phenolic acids, with catechin as the predominant flavonoid. Notably, BRR treatment led to elevated concentrations of the bioavailable quercetin-3-rhamnoside in 48-h-old sprouts, while 15-day-old plants grown under PFL conditions showed a significant accumulation of the sulfated quercetin-3-sulfate. Real-time PCR analysis demonstrated that BRR upregulated the expression of flavonoid biosynthesis genes PAL, chalcone synthase (CHS), and chalcone isomerase (CHI) in sprouts, whereas PFL treatment induced higher expression of these genes, as well as cinnamate 4-hydroxylase (C4H), in aerial parts. These findings suggest that targeted light treatments, particularly blue and red LED light, can enhance the accumulation of bioavailable quercetin-3-rhamnoside during T. persica germination and sprouts exhibit higher levels of flavonoids and phenolic acids than aerial parts during different vegetative growth stages.

Clinical significance of determining the level of citrulline in children who have undergone intestinal resection during the neonatal period

Citrulline is an amino acid that is not found in protein and does not enter the body through food, enteral or parenteral nutrition. It is instead synthesized exclusively by enterocytes. Therefore, citrulline concentrations in the blood can reflect the process of biosynthesis in the intestines and are directly related to changes in the function of these cells.Purpose. The aim of the study was to assess citrulline levels in children who had undergone intestinal resection during the neonatal period.Material and methods. The study included 22 children with short bowel syndrome, diagnosed by a doctor (main group; average age 3.7 months). We also included 20 children who had undergone surgery to remove less than 50% of their intestine but did not have short bowel syndrome (comparison group; average age 3 months 1 day). The control group consisted of 23 healthy children who had never had surgery on their gastrointestinal tract and did not have any congenital malformations or clinical signs of enteropathy (average age 5 months 3 days). High performance liquid chromatography-mass spectrometry (HPLC-MS) was used to measure the level of citrulline.Results. The average level of citrulline in the blood of children in the main group was 18 μmol/l (95% CI 10–22.8), in the comparison group – 17.5 μmol/l (95% CI 16–21), in the control group – 26 μmol/l (95% CI 22–3). Citrulline levels in patients who underwent surgery in the neonatal period were significantly different from those of children in the control group There were no differences in the content of citrulline between the main and comparison groups, depending on whether the colon was preserved or not and whether the ileocecal valve was present (p&gt;0.05).Conclusion. An increase in citrulline level in the blood of children receiving therapy with a glucagon-like peptide-2 analog indicates the possibility of using this marker to monitor changes in intestinal absorption associated with physiological or pharmacological adaptation of the intestine.

Enhancing agrochemical delivery in citrus leaves with geraniol: a promising strategy for controlling huanglongbing (HLB).

Citrus huanglongbing (HLB) is a devastating disease in citrus, caused by Candidatus Liberibacter asiaticus (CLas), which primarily resides in the phloem where chemicals cannot effectively reach, posing a significant challenge in controlling HLB. To address these challenges, plant essential oils (EOs), widely used as transdermal enhancers and known for their benefits for plant tissues, were investigated for their potential to enhance chemical permeation. In this study, seven EOs - eugenol, carvacrol, eucalyptol, geraniol, linalool, cinnamaldehyde, and d-limonene - were evaluated for their potential to enhance chemical penetration into citrus leaves. Preliminary screening with tracer methods revealed that geraniol provided the greatest enhancement of permeation, achieving the deepest and broadest distribution into the spongy tissue of the citrus leaf. The permeation levels of rhodamine B and crystal violet through the cuticular layer were elevated from 0.55 mg L-1 and 0.11 mg L-1 to 20.04 mg L-1 and 7.14 mg L-1, respectively, by the addition of geraniol. Further validation through high-performance liquid chromatography-mass spectrometry (HPLC-MS) confirmed that the penetration of fludioxonil and tetracycline into citrus leaf tissues was enhanced by 15.07-fold and 20.43-fold, respectively. In planta experiments confirmed that adding geraniol to tetracycline and oxytetracycline via foliar application eliminated CLas bacteria in HLB-affected citrus more efficiently than controls. Our study underscores the potential of geraniol as an effective EO-based permeation enhancer for combating HLB. By significantly improving the delivery and efficacy of bactericidal treatments, geraniol offers a promising strategy for managing this catastrophic citrus disease, potentially transforming the approach to controlling HLB. © 2024 Society of Chemical Industry.

Lingguizhugan decoction alleviates gestational diabetes mellitus by modulating the PI3K-AKT pathway and oxidative stress: Network pharmacology and experimental evidence.

The Lingguizhugan decoction (LGZGD) is a promising traditional Chinese medicine for the treatment of gestational diabetes mellitus (GDM). However, its bioactive compounds and therapeutic mechanisms remain unknown. The main chemical composition of LGZGD was analyzed by high-performance liquid chromatography-mass spectrometry (HPLC-MS). Furthermore, the underlying mechanisms of LGZGD against GDM were elucidated through network pharmacology and molecular docking. The therapeutic efficacy and targets of LGZGD were further confirmed via an in vitro GDM model (high glucose [HG]-treated HTR-8/SVneo cells). Four compounds of LGZGD, namely, cinnamaldehyde, glycyrrhizic acid, 2-atractylenolide, and pachymic acid, were detected. A total of 26 targets for LGZGD treating GDM were obtained, which were mainly involved in oxidative stress and the PI3K-AKT signaling pathway. The protein-protein interaction (PPI) network unveiled that AKT1, TLR4, TP53, and NOS3 were hub therapeutic targets. Molecular docking showed that these targets had strong affinity with key compounds. In vitro experiments confirmed that LGZGD treatment promoted HG-induced cell viability, migration, and invasion ability while inhibited the apoptosis rate and oxidative stress. Mechanically, western blot revealed that LGZGD may protect HG-treated cells by activating the PI3K-AKT pathway and suppressing TLR4 expression. Our study preliminarily explored the mechanism of LGZGD in GDM treatment, providing a scientific basis for the clinical application of LGZGD.

Automated-Screening Oriented Electric Sensing of Vitamin B1 Using a Machine Learning Aided Solid-State Nanopore.

Micronutrient detection and identification at the single-molecule level are paramount for both clinical and home diagnostics. Analytical tools such as high-performance liquid chromatography and liquid chromatography-tandem mass spectrometry have been widely used but include a high instrument cost and prolonged analysis time. Here, as a model system, by merging nanopore signatures with machine learning algorithms, we propose an automated electric sensing strategy to identify vitamin B1 and its phosphorylated derivatives with good accuracy. Further, the relationship between vitamin B1 dynamics and nanopore signatures is examined. To understand the machine-decision-making process, Shapley additive explanations are made. Using a machine learning aided solid-state nanopore, we pave the way for next-generation micronutrient detection.

Genomic Features and Antimicrobial Activity of Phaeobacter inhibens Strains from Marine Biofilms.

Members of the genus Phaeobacter are widely distributed in the marine environment and are known for their ability to produce tropodithietic acid (TDA). Studies investigating the genomic and metabolic features of Phaeobacter strains from marine biofilms are sparse. Here, we analyze the complete genomes of 18 Phaeobacter strains isolated from biofilms on subtidal stones, with the aim of determining their potential to synthesize secondary metabolites. Based on whole-genome comparison and average nucleotide identity calculation, the isolated bacteria are classified as novel strains of Phaeobacter inhibens. Further analysis reveals a total of 153 biosynthetic gene clusters, which are assigned to 32 gene cluster families with low similarity to previously published ones. Complete TDA clusters are identified in 14 of the 18 strains, while in the other 4 strains the TDA clusters are rather incomplete and scattered across different chromosome and plasmid locations. Phylogenetic analysis suggests that their presence or absence may be potentially attributed to horizontal gene transfer. High-performance liquid chromatography-mass spectrometry analysis demonstrates the production of TDA in all the examined strains. Furthermore, the Phaeobacter strains have strong antibacterial activity against the pathogenic strain Vibrio owensii ems001, which is associated with acute hepatopancreatic necrosis in South American white shrimp. Altogether, this study ameliorates our knowledge of marine biofilm-associated Phaeobacter and offers new avenues for exploiting marine antimicrobial agents.

Roseoside Is a Bioactive Compound in Kirengeshoma koreana Nakai Extract with Potent In Vitro Antiviral Activity Against Hepatitis C Virus.

Hepatitis C virus (HCV) is a pathogen that causes cirrhosis and hepatocellular carcinoma through chronic hepatitis C. This study focused on the anti-HCV activity of a 70% ethanol extract of Kirengeshoma koreana Nakai (KKE) and its bioactive chemical constituent(s). The KKE and its n-butanol (n-BuOH) fraction induced a significant reduction in HCV RNA levels without inducing cytotoxicity. A high-performance liquid chromatography-mass spectrometry (HPLC-MS) analysis revealed the presence of roseoside in the n-butanol fraction of the KKE, which inhibited HCV RNA replication in a concentration- and time-dependent manner without exerting cytotoxicity. Consistent with in silico molecular docking analysis data, roseoside targets and inhibits HCV NS5A/B replicase. Collectively, our findings demonstrate that roseoside is a chemical constituent in KKE that interferes with HCV replication by targeting NS5A/B replicase.

Comparison of the potential risk from arsenic exposure when eating fish grown in natural and artificial conditions, using the example of rainbow trout

Introduction. Due to the increasing share of fish and fish products in the human diet, hygienic problems are also increasing, especially with the growing of aquaculture sector. The task of determining the level of arsenic toxicity is becoming extremely important. Although the toxicity mechanisms of inorganic arsenic are generally well understood, the impact of speciation of its other (organic) compounds on the likelihood of carcinogenic and non-carcinogenic effects has been largely unexplored. Material and methods. The objects of study were samples of fillet and caviar of salmon fish – rainbow trout (wild and aquaculture). The quantitative determination of arsenic-containing substances was carried out using high-performance liquid chromatography-mass spectrometry with inductively coupled plasma. The samples were prepared using microwave methods. The risk level was calculated based on recommendations R 2.1.10.3968–23. Results. As the result of this work the information about concentration of arsenic (organic and inorganic compounds) in fillets and caviar of fish from the salmon family - rainbow trout was shown. It has been established that the content of inorganic and organic arsenic compounds practically does not differ, depending of the habitat of trout. The levels of carcinogenic and non-carcinogenic risk of inorganic arsenic were calculated. Limitations. Determination of arsenic concentration was carried out in rainbow trout, which had been grown in natural and artificial conditions. Conclusion. It has been established that inorganic arsenic makes the greatest contribution to the health risk. However, the magnitude of the risk does not exceed the acceptable sanitary and hygienic standards, while the conditions for growing fish have practically no effect on these values.

Development of microflow ultra high performance liquid chromatography-mass spectrometry metabolomic assays for analysis of mammalian biofluids

Introduction and objectivesThe application of untargeted metabolomics assays using ultra high performance liquid chromatography-mass spectrometry (UHPLC-MS) to study metabolism in biological systems including humans is rapidly increasing. In some of these studies there is a requirement to collect and analyse low sample volumes of biofluids (e.g. tear fluid) or low cell and tissue mass samples (e.g. tissue needle biopsies). The application of microflow, capillary or nano liquid chromatography (≤ 1.0 mm column internal diameter (i.d.)) theoretically should accomplish a higher assay sensitivity compared to analytical liquid chromatography (2.1–5.0 mm column internal diameter). To date, there has been limited research into microflow UHPLC-MS assays that can be applied to study samples of low volume or mass.MethodsThis paper presents three complementary UHPLC-MS assays (aqueous C18 reversed-phase, lipidomics C18 reversed-phase and Hydrophilic Interaction Liquid Chromatography (HILIC)) applying 1.0 mm internal diameter columns for untargeted metabolomics. Human plasma and urine samples were applied for the method development, with porcine plasma, urine and tear fluid used for method assessment. Data were collected and compared for columns of the same length, stationary phase and stationary phase particle size but with two different column internal diameters (2.1 mm and 1.0 mm).Results and conclusionsAll three assays showed an increase in peak areas and peak widths when applying the 1.0 mm i.d. assays. HILIC assays provide an advantage at lower sample dilutions whereas for reversed phase (RP) assays there was no benefit added. This can be seen in the validation study where a much higher number of compounds were detected in the HILIC assay. RP assays were still appropriate for small volume samples with hundreds of compounds being detected. In summary, the 1.0 mm i.d. column assays are applicable for small volume samples where dilution is required during sample preparation.

Identification of metabolic biomarkers in idiopathic pulmonary arterial hypertension using targeted metabolomics and bioinformatics analysis

Pulmonary arterial hypertension (PAH) is a life-threatening disease with a poor prognosis, and metabolic abnormalities play a critical role in its development. This study used metabolomics, machine learning algorithms and bioinformatics to screen for potential metabolic biomarkers associated with the diagnosis of PAH. In this study, plasma samples were collected from 17 patients diagnosed with idiopathic pulmonary arterial hypertension (IPAH) and 20 healthy controls. Plasma metabolomic profiling was performed by high-performance liquid chromatography-mass spectrometry. Gene profiles of PAH patients were obtained from the GEO database. Key differentially expressed metabolites (DEMs) and metabolism-related genes were subsequently identified using machine learning algorithms. Twenty differential plasma metabolites associated with IPAH were identified (VIP score > 1 and p < 0 0.05), and enrichment analysis revealed the arginine biosynthesis pathway as the most altered pathway. Using machine learning models, including least absolute shrinkage and selection operator (LASSO), random forest (RF) and support vector machine (SVM), we extracted key metabolites that correlated with clinical phenotypes. Our results suggested that five metabolites, kynurenine, homoserine, tryptophan, AMP, and spermine, are potential biomarkers for IPAH. Bioinformatics analysis also identified 3 metabolism-related genes, MAPK6, SLC7A11 and CDC42BPA, that are strongly correlated with pulmonary hypertension, demonstrating strong predictive power and clinical relevance. Our findings revealed some key genes associated with metabolism in PH, and provided crucial information about complex metabolic reprogramming signals and may lead to the identification of useful metabolic biomarkers for the diagnosis of PAH.

Drug Use during Incarceration: A Comprehensive Quality and Prevalence Study in Three Danish Prisons

Background Drug use in Danish prisons has previously not been investigated in detail using confirmatory, laboratory analysis. The objective of the present quality study, initiated by the Danish Prison and Probation Service, was to i) evaluate the performance of an initial, on-site drug screening strategy; ii) gain insights into emerging drug trends; and iii) suggest evidence-based strategies for future drug testing. Methods Over a two-year period, routine urine samples (n = 1952 from 710 inmates) from three Danish prisons were subjected to comprehensive drug testing. Immunoassay screening was conducted on-site. A parallel sample aliquot was forwarded to laboratory analysis by confirmatory methods: High-performance liquid chromatography and tandem mass spectrometry (LC-MS/MS) and gas chromatography and mass spectrometry (GC-MS) targeting 56 drugs-of-abuse/medical drugs, 26 key metabolites, 41 new psychoactive substances (NPS), including specific biomarkers for heroin, crack cocaine, or ethanol (a total of 123 target analytes/sample). Results The on-site immunoassay method showed a sensitivity from 66 to 100%; specificity was above 98%; accuracy was above 95%. Laboratory analysis detected compounds not screened for including tramadol, oxycodone, buprenorphine, ketamine, MDMA, 4-fluoroamphetamine, and GHB. The prevalence of drug use was in the order: cannabis > ethanol > cocaine > benzodiazepines > amphetamine. Conclusion The performance of the immunoassay was found acceptable; however, the screening program was inadequate for detecting other significant substances. Based on these findings, a broader screening method will be implemented in future at Danish Prisons to minimize false negative results. The data did not indicate a trend of using NPS in Danish prisons.

Downstream effects of the pandemic? Spatiotemporal trends of quaternary ammonium compounds in suspended particulate matter of German rivers

During the SARS-CoV-2 pandemic, the preventive use of antimicrobials such as quaternary ammonium compounds (QACs) surged worldwide. As cationic and surface-active biocides, QACs are only partly removed during wastewater treatment and may cause adverse ecological effects in the downstream environment. To understand the environmental consequences of increased disinfectant use during the pandemic, we investigated spatiotemporal QAC concentration trends in the suspended particulate matter (SPM) of three diverse German rivers. Covering pooled annual SPM samples from 2006–2021 and monthly samples from 2018–2021 collected by the German Environmental Specimen Bank, 31 QACs were quantified by high performance liquid chromatography–mass spectrometry. ∑QAC concentrations in annual samples differed by more than tenfold between rivers in the order Saar (average 6.7 µg/g) > Rhine (0.9 µg/g) > Mulde (0.3 µg/g). The strongest potential pandemic imprint was however observed in the Mulde (+67 %) and Rhine (+22 %). Besides pandemic dynamics, also seasonal variation and mineral content of SPM tentatively affected QAC concentrations. Exceedance of predicted no-effect concentrations for sediment suggest ecotoxicological risks for long-chained QACs already before the pandemic. Our study thus highlights the importance of monitoring the environmental effects of antimicrobial use during pandemics and calls for an urgent minimization of non-essential applications.

Activation of persulfate by MOF-derived MnFeOx to efficiently degrade sulfadiazine: Synergistic effects from free radicals and singlet oxygen

In this study, a facile hydrothermal approach was used to synthesize the MOFs materials [Mn-doped MIL-101(Fe)]. These materials were subsequently subjected to high temperature annealing process via calcination to generate the MOFs derivatives (MnFeOx). The synthesized materials were utilized for the removal of sulfadiazine (SDZ) removal through activated peroxymonosulfate (PMS). At the optimum Mn doping, the removal ratio of SDZ could reach 98.9 % (at natural pH) within 20 min. In addition, the study investigated the influence of PMS dosage, catalyst dosage, initial pH, various inorganic anions, and humic acid (HA) on the degradation of SDZ. The activation processes of both free and non-free radicals in the MnFeOx/PMS system were examined by free radical quenching experiments and electron paramagnetic resonance (EPR) techniques. The analysis of the degradation products of SDZ was conducted by using high performance liquid chromatography-mass spectrometry (HPLC-MS), and the breakdown pathways of SDZ in the MnFeOx/PMS system were proposed.