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In order to investigate the key nodes affecting the changes of Yixian dark tea flavor and quality during the Pile fermentation, in this study, the sensory quality and non-volatile profiles of Yixian dark tea samples at different pile fermentation periods (0, 3, 6, 9, 12, 15, 18 h) were investigated by sensory evaluation, amino acids analyzer, liquid chromatography and liquid chromatography mass spectrometry. The results showed that the intensity of bitterness in dark tea infusions declined after 9 h of pile fermentation, along with the increasing intensities of astringency, thickness and stale-mellow. Total catechins showed a significant decrease in concentrations during pile fermentation. In contrast, the concentration of gallic acid increased. Concentration of caffeine reduced by 14.7% compared to the original concentration after 18 h of pile fermentation. Total amino acids concentration showed an increasing tendency in the later period of pile fermentation. Concentrations of flavonoids and its mono-glycosides increased after 9-12 hours, while its di-glycosides showed an increasing tendency after 12 hours of pile fermentation. The analysis of key taste substances (VIP>1, DOT>1) showed that myricetin-3-O-galactoside, astragalin, isoquercitrin, quercetin-3-O-rutinose, epigallocatechin, epicatechin, epigallocatechin gallate, and caffeine were the most important flavor presenting substances affecting the quality changes of Yixian dark tea. Among them, the content of the first three glycosides increased significantly from 9-12 hours of stacking; The contents of epigallocatechin and epicatechin were significantly reduced during 9-12 hours. Epigallocatechin gallate and caffeine showed a significant decrease after pile fermentation; Quercetin-3-O-rutinoside decreased significantly in the middle stage (9-12 hours) and increased significantly after 15 hours. It was an important factor in the increase of astringency of tea broth in the late stage of pile fermentation. This study give a theoretical basis for improving the tea processing of Yixian dark tea and understanding the formation mechanism of its key flavor. [ABSTRACT FROM AUTHOR]

In order to investigate the key nodes affecting the changes of Yixian dark tea flavor and quality during the Pile fermentation, in this study, the sensory quality and non-volatile profiles of Yixian dark tea samples at different pile fermentation periods (0, 3, 6, 9, 12, 15, 18 h) were investigated by sensory evaluation, amino acids analyzer, liquid chromatography and liquid chromatography mass spectrometry. The results showed that the intensity of bitterness in dark tea infusions declined after 9 h of pile fermentation, along with the increasing intensities of astringency, thickness and stale-mellow. Total catechins showed a significant decrease in concentrations during pile fermentation. In contrast, the concentration of gallic acid increased. Concentration of caffeine reduced by 14.7% compared to the original concentration after 18 h of pile fermentation. Total amino acids concentration showed an increasing tendency in the later period of pile fermentation. Concentrations of flavonoids and its mono-glycosides increased after 9~12 hours, while its di-glycosides showed an increasing tendency after 12 hours of pile fermentation. The analysis of key taste substances (VIP>1, DOT>1) showed that myricetin-3-O-galactoside, astragalin, isoquercitrin, quercetin-3-O-rutinose, epigallocatechin, epicatechin, epigallocatechin gallate, and caffeine were the most important flavor presenting substances affecting the quality changes of Yixian dark tea. Among them, the content of the first three glycosides increased significantly from 9~12 hours of stacking; The contents of epigallocatechin and epicatechin were significantly reduced during 9~12 hours. Epigallocatechin gallate and caffeine showed a significant decrease after pile fermentation; Quercetin-3-O-rutinoside decreased significantly in the middle stage (9~12 hours) and increased significantly after 15 hours. It was an important factor in the increase of astringency of tea broth in the late stage of pile fermentation. This study give a theoretical basis for improving the tea processing of Yixian dark tea and understanding the formation mechanism of its key flavor.

Abstract Prognostic tests and treatment approaches for optimized clinical care of prostatic neoplasms are an unmet need. Prostate cancer (PCa) and derived extracellular vesicles (EVs) proteome changes occur during initiation and progression of the disease. PCa tissue proteome has been previously characterized, but screening of tissue samples constitutes an invasive procedure. Consequently, we focused this study on liquid biopsies, such as urine samples. More specifically, urinary small extracellular vesicle and particles proteome profiles of 100 subjects were analyzed using liquid chromatography coupled to high-resolution mass spectrometry (LC–MS/MS). We identified 171 proteins that were differentially expressed between intraductal prostate cancer/cribriform (IDC/Crib) and non-IDC/non-Crib after correction for multiple testing. However, the strong correlation between IDC/Crib and Gleason Grade complicates the disentanglement of the underlying factors driving this association. Nevertheless, even after accounting for multiple testing and adjusting for ISUP (International Society of Urological Pathology) grading, two proteins continued to exhibit significant differential expression between IDC/Crib and non-IDC/non-Crib. Functional enrichment analysis based on cancer hallmark proteins disclosed a clear pattern of androgen response down-regulation in urinary EVs from IDC/Crib compared to non-IDC/non-Crib. Interestingly, proteome differences between IDC and cribriform were more subtle, suggesting high proteome heterogeneity. Overall, the urinary EV proteome reflected partly the prostate pathology.

Chardonnay and Sauvignon blanc wines aged in oak wood barrels with low and medium tannin potentials were discriminated for their abilities to resist against oxidation during bottle storage. The oak wood tannin potential was positively correlated to wines antioxidant capacity after 2 and 4 years of bottle aging. Untargeted molecular analysis revealed that the Sauvignon blanc metabolome was more affected by the tannin potential than the Chardonnay. Supervised statistical analysis highlighted the extensive oak wood contribution to the wine chemical fingerprints. Wines aged in barrel of medium tannin potential were associated with higher concentrations in antioxidant compounds such as dipeptides. Moreover, quantitative differences were observed between oak barrel derived volatile compounds. Sauvignon blanc volatile thiols appeared to decrease during bottle aging, regardless of the oak tannin potential. This study highlights the post bottling positive impact of oak wood barrel aging on wines oxidative stability, related to oak barrel tannin potential.

The aim of this study was to assess the nutraceutical qualities of extra virgin olive oil (EVOO) samples obtained from three Sicilian olive cultivars: Nocellara, Biancolilla, and Cerasuola. We also evidenced the relationship among biophenols, base parameters and panel test scores, and evaluated the stability of the biophenols in EVOO. The assessment also took into consideration variations in olive harvesting periods and the influence of four different milling methods. A statistical analysis of the collected data revealed that the cultivar and harvesting period were the primary factors influencing the bio-phenol content, while the milling methods employed did not significantly affect the levels of biophenols in the oils. The panel test results were also illuminating as they were strongly related to the cultivar and polyphenol content. Following the criteria outlined in EC Regulation 432/2012, we selected three samples, each representing one of the cultivars, which exhibited the highest bio-phenol content to evaluate the biophenol stability during a time span of 16 months. [ABSTRACT FROM AUTHOR]

This document presents the results of a study comparing the accuracy of a cortisol immunoassay to LC-MS/MS. The study found a significant positive bias in the cortisol immunoassay compared to LC-MS/MS, but could not identify the cause of this difference. However, the study suggests that despite the bias, the cortisol immunoassay may still be considered for adrenal vein sampling (AVS) due to the minimal impact on clinical decision making. The study recommends careful consideration of the positive bias for other indications. The document concludes with acknowledgments to the individuals who contributed to the analysis of the samples. [Extracted from the article]

The feasibility of using a 26‐1 fractional factorial design to screen the relative importance of six water quality and operational factors in the removal of microcystin‐LR (MC‐LR) by powdered activated carbon (PAC) was evaluated through jar testing. The factors were: PAC type, PAC dose, total organic carbon (TOC) concentration, turbidity, alum dose, and timing of PAC versus coagulant application. Follow‐up tests were performed to examine the interaction of PAC dose and TOC concentrations. All MC‐LR analyses were performed by enzyme linked immunosorbent assay (ELISA) and liquid chromatography coupled with tandem mass spectrometry (LC/MS/MS). The top three effect magnitudes were the same by ELISA and LC/MS/MS:PAC dose > PAC type > PAC application time. Correlation coefficients between removals estimated by ELISA and LC/MS/MS were >0.9 (p «.05). With both methods, the effects of PAC type and dose were found to be markedly larger than the other factors. The follow‐up tests indicated a greater impact of PAC dose at higher natural organic matter concentrations. Factorial designs are not commonly used to plan drinking water jar test experiments. The results generated in this study were plausible with respect to the existing body of adsorption knowledge, thus helping to demonstrate the feasibility of the factorial approach. [ABSTRACT FROM AUTHOR]

Structure identification of drug degradation products is one of the key components of any pharmaceutical product development. Knowledge-based in-silico tools are widely used to predict the probable degradation and excipient interaction products. However, multiple structures of the same masses are predicted through this software, which makes identification of the correct/exact structure of the degradation product difficult. In this study, the utilization of a simple yet powerful analytical tool, LC-MS HDX was explored for improving the predictive capability of the in-silico tool. For the same, 5 drugs were selected as representative of pharmaceutical development workflow and subjected to a stress degradation study as per ICH regulatory requirements. There were in total 55 degradation products, for which all possible structures were predicted through Zeneth® software. The number of labile hydrogens in each of the 55 products was obtained through a simple LC-MS HDX method. While scrutinizing predicted structures with the number of labile hydrogens, many false positive predictions could be ruled out. Out of 55 degradation products, this approach has helped in improving the predictability of 45 products corresponding to ∼82% cases. It is simple to adopt for the industry by simply amalgamating analytical practices with AI prediction. [ABSTRACT FROM AUTHOR]

Abstract Background The inclusion of dexmedetomidine (DEX) within a balanced general anaesthesia protocol is effective in improving the clinical outcome and recovery quality of anaesthesia in horses. This study aimed to determine the pharmacokinetic profile of DEX following repeated subcutaneous (SC) administration at 2 µg/kg every 60 min till the end of the procedure in comparison to intravenous constant rate infusion (CRI) at 1 µg/kg/h in anaesthetized horses undergoing diagnostic procedures up to the end of the diagnostic procedure. Results In the CRI and SC groups DEX maximum concentrations (Cmax) were 0.83 ± 0.27 ng/mL and 1.14 ± 0.71 ng/mL, respectively, reached at a time (Tmax) of 57.0 ± 13.4 min and 105.5 ± 29.9 min. Mean residence time to the last measurable concentration (MRTlast) was 11.7 ± 6.2 and 55.8 ± 19.7 min for the CRI group and SC groups, respectively. The apparent elimination half-life was 18.0 ± 10.0 min in the CRI group and 94.8 ± 69.8 min for the SC group, whereas the area under the curve (AUC0-last) resulted 67.7 ± 29.3 and 83.2 ± 60.5 min*ng/mL for CRI and SC group, respectively. Clearance was 16.26 ± 8.07 mL/min/kg for the CRI group. No signs of adverse effects were recorded in both groups. Conclusions The pharmacokinetic profile of DEX following repeated SC administration in anaesthetized horses was comparable to intravenous CRI administration during the intranaesthetic period and beneficial during the recovery phase from general anaesthesia. The SC route could be considered as an alternative to CRI for improving the recovery quality of equine patients undergoing general anaesthesia.

Exometabolomics is an approach to assess how microorganisms alter, or react to their environments through the depletion and production of metabolites. It allows the examination of how soil microbes transform the small molecule metabolites within their environment, which can be used to study resource competition and cross-feeding. This approach is most powerful when used with defined media that enable tracking of all metabolites. However, microbial growth media have traditionally been developed for the isolation and growth of microorganisms but not metabolite utilization profiling through Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). Here, we describe the construction of a defined medium, the Northen Lab Defined Medium (NLDM), that not only supports the growth of diverse soil bacteria but also is defined and therefore suited for exometabolomic experiments. Metabolites included in NLDM were selected based on their presence in R2A medium and soil, elemental stoichiometry requirements, as well as knowledge of metabolite usage by different bacteria. We found that NLDM supported the growth of 108 of the 110 phylogenetically diverse (spanning 36 different families) soil bacterial isolates tested and all of its metabolites were trackable through LC-MS/MS analysis. These results demonstrate the viability and utility of the constructed NLDM medium for growing and characterizing diverse microbial isolates and communities.

Introduction: Doping and steroid use represent a serious threat to animal health and can even lead to their untimely and painful death. However, doping is an acute problem in today's animal racing world, particularly in camel racing. Testosterone and its ten esters (benzoate, valerate, isocaproate, hexahydrobenzoate, decanoate, undecanoate, laurate, enanthate, cypionate, and caproate) are of utmost importance, because when they are administered to animals it is difficult to measure them efficiently. The levels of testosterone and its esters in camels and other animals are typically determined using urine and blood tests. The aim of this study was to develop and validate a liquid chromatographic–mass spectrometric (LC-MS/MS) method to determine testosterone esters in camel hair, and to apply the validated method to determine testosterone esters in collected samples. To our knowledge, this is the first report of such research. Results and Discussion: The levels of testosterone and its ten derivatives, along with the cortisol-D4 internal standard, were optimised for LC–MS/MS analysis; however, only testosterone along with its seven esters (namely benzoate, valerate, isocaproate, hexahydrobenzoate, decanoate, undecanoate and laurate) could be validated in camel hair. Only five testosterone esters could be determined in camel hair samples; the concentrations were obtained as 10.5–14.9 pg/mg for valerate (in three camels), 12.5–151.6 pg/mg for hexahydrobenzoate (in six camels), 4.8–32.1 pg/mg for laurate (in five camels), 5.1 pg/mg decanoate (in one camel), and 8.35–169 pg/mg for testosterone (in all 24 camels). Interestingly, the three racing camels displayed high concentrations of testosterone (59.2–169 pg/mg, all three camels), laurate (4.8–14.5 pg/mg, two camels), hexahydrobenzoate (116 pg/mg, one camel), decanoate (5.1 pg/mg, one camel), and valerate (11.7 pg/mg, one camel). Methods: Camel hair samples were collected from 21 non-racing dromedary camels along with three racing camels in Al Ain, UAE; these were decontaminated, pulverised, sonicated, and extracted prior to analysis. An LC–MS/MS method was employed to determine the levels of testosterone esters in the hair samples. Conclusions: This novel camel-hair test procedure is accurate, sensitive, rapid, and robust. The findings reported in this study could be significant to evaluate racing camels for suspected doping offenses. Further controlled testosterone supplementation studies are required to evaluate individual esters' effects on camel health and diseases and on performance enhancement levels. This new hair test could promote further studies in doping control, toxicology, and pharmacology, as well as having other clinical applications relating to camel health, injury, and disease. [ABSTRACT FROM AUTHOR]

This study aimed to determine whether leaf extracts from seven Eruca vesicaria subsp. sativa cultivars and their biochemically active compounds (glucosinolates and downstream-derived products) inhibit mycelia growth of three well-known pathogenic oomycetes, Phytopythium chamaehyphon, Phytopythium vexans and Phytophthora citrophthora; being the most significant in the development of Kiwifruit Vine Decline Syndrome (KVDS). Leaf extract quantity of 10, 20 and 30 mgwereinoculated in Petridish (90mmØ, each 22mLof liquidmedium-Potato Dextrose Agar), for in vitro bioassays. A pathogen plug was placed in the centre of each plate and the Oomycota colony perimeter was marked 5 days after inoculation. Radial colony growth was measured from 4 marks per plate 5, 10, and 15 days after inoculation, further elaborated with Image J software image analysis. Growthrates forall strainswereinhibitedbyaround67%after15days. This wasmostpronouncedwhenapplying thehighest concentrationof leaf extract. By using Liquid Chromatography Mass Spectrometry (LC-MS) and Gas Chromatography Mass Spectrometry (GC-MS), fifteen glucosinolate compounds, of which glucosativin was found in the highest quantity, were identified. Concentrations of hydrolysis products produced by leaves (erucin and sativin) were also investigated, and were significantly associated with colony radial growth, especially towards Pp. chamaehyphon and Pp. vexans. Three downstream products of glucosinolates (two pure isothiocyanates, AITC and PEITC; and one indole I3C; all commonly present in Brassicaceae) were also tested, and a statistically significant inhibition of growth was observed at the highest concentration (0.6 µL). [ABSTRACT FROM AUTHOR]

Background: The inclusion of dexmedetomidine (DEX) within a balanced general anaesthesia protocol is effective in improving the clinical outcome and recovery quality of anaesthesia in horses. This study aimed to determine the pharmacokinetic profile of DEX following repeated subcutaneous (SC) administration at 2 µg/kg every 60 min till the end of the procedure in comparison to intravenous constant rate infusion (CRI) at 1 µg/kg/h in anaesthetized horses undergoing diagnostic procedures up to the end of the diagnostic procedure. Results: In the CRI and SC groups DEX maximum concentrations (Cmax) were 0.83 ± 0.27 ng/mL and 1.14 ± 0.71 ng/mL, respectively, reached at a time (Tmax) of 57.0 ± 13.4 min and 105.5 ± 29.9 min. Mean residence time to the last measurable concentration (MRTlast) was 11.7 ± 6.2 and 55.8 ± 19.7 min for the CRI group and SC groups, respectively. The apparent elimination half-life was 18.0 ± 10.0 min in the CRI group and 94.8 ± 69.8 min for the SC group, whereas the area under the curve (AUC0-last) resulted 67.7 ± 29.3 and 83.2 ± 60.5 min*ng/mL for CRI and SC group, respectively. Clearance was 16.26 ± 8.07 mL/min/kg for the CRI group. No signs of adverse effects were recorded in both groups. Conclusions: The pharmacokinetic profile of DEX following repeated SC administration in anaesthetized horses was comparable to intravenous CRI administration during the intranaesthetic period and beneficial during the recovery phase from general anaesthesia. The SC route could be considered as an alternative to CRI for improving the recovery quality of equine patients undergoing general anaesthesia. [ABSTRACT FROM AUTHOR]

The aim of this study was to assess the nutraceutical qualities of extra virgin olive oil (EVOO) samples obtained from three Sicilian olive cultivars: Nocellara, Biancolilla, and Cerasuola. We also evidenced the relationship among biophenols, base parameters and panel test scores, and evaluated the stability of the biophenols in EVOO. The assessment also took into consideration variations in olive harvesting periods and the influence of four different milling methods. A statistical analysis of the collected data revealed that the cultivar and harvesting period were the primary factors influencing the bio-phenol content, while the milling methods employed did not significantly affect the levels of biophenols in the oils. The panel test results were also illuminating as they were strongly related to the cultivar and polyphenol content. Following the criteria outlined in EC Regulation 432/2012, we selected three samples, each representing one of the cultivars, which exhibited the highest bio-phenol content to evaluate the biophenol stability during a time span of 16 months.

This study aimed to determine whether leaf extracts from seven Eruca vesicaria subsp. sativa cultivars and their biochemically active compounds (glucosinolates and downstream-derived products) inhibit mycelia growth of three well-known pathogenic oomycetes, Phytopythium chamaehyphon, Phytopythium vexans and Phytophthora citrophthora; being the most significant in the development of Kiwifruit Vine Decline Syndrome (KVDS). Leaf extract quantity of 10, 20 and 30 mg were inoculated in Petri dish (90 mm Ø, each 22 mL of liquid medium – Potato Dextrose Agar), for in vitro bioassays. A pathogen plug was placed in the centre of each plate and the Oomycota colony perimeter was marked 5 days after inoculation. Radial colony growth was measured from 4 marks per plate 5, 10, and 15 days after inoculation, further elaborated with Image J software image analysis. Growth rates for all strains were inhibited by around 67% after 15 days. This was most pronounced when applying the highest concentration of leaf extract. By using Liquid Chromatography Mass Spectrometry (LC-MS) and Gas Chromatography Mass Spectrometry (GC-MS), fifteen glucosinolate compounds, of which glucosativin was found in the highest quantity, were identified. Concentrations of hydrolysis products produced by leaves (erucin and sativin) were also investigated, and were significantly associated with colony radial growth, especially towards Pp. chamaehyphon and Pp. vexans. Three downstream products of glucosinolates (two pure isothiocyanates, AITC and PEITC; and one indole I3C; all commonly present in Brassicaceae) were also tested, and a statistically significant inhibition of growth was observed at the highest concentration (0.6 µL).

[Display omitted] Multi-attribute methods employing mass spectrometry are applied throughout the biopharmaceutical industry for product and process characterization purposes but are not yet widely accepted as a method for batch release and stability testing under the good manufacturing practice (GMP) regime, due to limited experience and level of comfort with the technical, compliance and regulatory aspects of its implementation at quality control (QC) laboratories. This article is the second part of a two-tiered publication aiming at providing guidance for implementation of the multi-attribute method by peptide mapping liquid chromatography mass spectrometry (MAM) in a QC laboratory. The first part [1] focuses on technical considerations, while this second part provides considerations related to GMP compliance and regulatory aspects. This publication has been prepared by a group of industry experts representing 14 globally acting major biotechnology companies under the umbrella of the European Federation of Pharmaceutical Industries and Associations (EFPIA) Manufacturing & Quality Expert Group (MQEG). [ABSTRACT FROM AUTHOR]

A highly sensitive method was developed for the determination of 17 nitrated and oxygenated polycyclic aromatic hydrocarbons (NPAHs and OPAHs) in PM2.5 using an ultrasound-assisted extraction (UAE)-online solid phase extraction (online SPE)-liquid chromatography-tandem mass spectrometry (LC/MS). The method underwent comprehensive optimization, encompassing UAE solvents and times, extract filtering, online SPE condition and LC/MS condition. Isomeric separation and detection of mononitro-PAHs, dinitro-PAHs, and OPAHs were achieved based on the proposed MS/MS fragmentation pathway in negative mode atmospheric pressure chemical ionization. Taking the advantage of online SPE, the method achieved lower limits of detection (NPAHs: 0.001 ~ 0.042 μg L−1, OPAHs: 0.01 ~ 0.038 μg L−1). Recovery ranged from 64% ~ 108% at three spiking levels (0.25, 1.0 and 10.0 μg L−1) with the relative standard deviations (RSDs) of 3% ~ 23%. This study represents first application of LC/MS methodology for the quantification of three carcinogenic dinitro-PAHs (1,3-dinitropyrene, 1,6-dinitropyrene, 1,8-dinitropyrene) in atmospheric PM2.5. The established method was successfully employed to determine the wide range concentrations of various NPAHs and OPAHs (∑NPAH: 9.4 ~ 303.3 pg m−3, ∑OPAHs: 125.2 ~ 1948.1 pg m−3) in PM2.5 samples collected in Beijing during the summer and winter seasons. [ABSTRACT FROM AUTHOR]

Filipendula ulmaria, commonly known as meadowsweet, is a wild herbaceous flowering plant that is widely distributed in Europe. A range of salicylic acid derivatives and flavonol glycosides have been previously associated with the antirheumatic and diuretic properties of F. ulmaria. In the present work, a hydroalcoholic extract from F. ulmaria aerial parts was extensively profiled using an efficient NMR-based dereplication strategy. The approach involves the fractionation of the crude extract by centrifugal partition chromatography (CPC), 13C NMR analysis of the fractions, 2D-cluster mapping of the entire NMR dataset, and, finally, structure elucidation using a natural metabolite database, validated by 2D NMR data interpretation and liquid chromatography coupled with mass spectrometry. The chemodiversity of the aerial parts was extensive, with 28 compounds unambiguously identified, spanning various biosynthetic classes. The F. ulmaria extract and CPC fractions were screened for their potential to enhance skin epidermal barrier function and skin renewal properties using in vitro assays performed on Normal Human Epidermal Keratinocytes. Fractions containing quercetin, kaempferol glycosides, ursolic acid, pomolic acid, naringenin, β-sitosterol, and Tellimagrandins I and II were found to upregulate genes related to skin barrier function, epidermal renewal, and stress responses. This research is significant as it could provide a natural solution for improving hydration and skin renewal properties. [ABSTRACT FROM AUTHOR]

In clinical practice, steroid measurements are performed mainly by direct, non-extraction immunoassays adapted to high throughput, automated immunoassay platforms and employing secondary calibrators. The accuracy of such steroid immunoassays is limited by cross-reactivity with structurally related steroids and nonspecific matrix interference as well as the metrological traceability of manufacturer supplied calibrators. The accuracy of steroid immunoassay calibrators has been little investigated by independent chemical methods. Steroid concentrations of 41 calibrators (4–6 replicates per calibrator) supplied by four manufacturers for use in testosterone (T), estradiol (E2), and progesterone (P4) commercial immunoassays were measured by ultra-pressure liquid chromatography-mass spectrometry (UPLC-MS). Among 14 non-zero T calibrators, six (43 %) deviated significantly from the label concentration with 29 % outside 20 % of it. Among 14 E2 calibrators, eight (57 %) deviated significantly, whereas seven (50 %) were outside 20 % of the label concentration. Among 11 P4 calibrators, eight (73 %) deviated significantly whereas four (36 %) were outside within 20 % of the label concentration. We conclude that inaccurate calibration of manufacturer's supplied standards may contribute to inaccuracy of commercial direct steroid immunoassays. [ABSTRACT FROM AUTHOR]

Fluoroquinolones and amphenicols are widely used as broad-spectrum and high-efficiency antibiotics in livestock and poultry production, but excessive drug residues frequently occur. Liquid chromatographic and liquid chromatographic mass spectrometry are the primary methods to detect the fluoroquinolones and amphenicols. According to the current domestic standard, these two veterinary drug residues can only be detected in milk simultaneously, and there are few reports in other matrices. Therefore, it is urgent to establish and optimize the simultaneous detection methods of fluoroquinolones and amphenicols in different matrices. This paper summarizes four standard pretreatment techniques such as liquid-liquid extraction, solid-phase extraction, QuEChERS method and accelerated solvent extraction, and discusses the application of hotspot nanomaterial adsorbent in detail. From the perspective of chromatography and chromatography mass spectrometry, the effects of different matrix conditions, instrument conditions and liquid phase conditions on the performance parameters of the method are summarized and discussed to provide a reference for the detection and supervision of veterinary drug residues in animal-derived foods.

Lotus rhizome starch (LRS) is an important traditional processed product of lotus rhizome in China due to its good taste and high nutritional value. However, adulterated lotus rhizome starches are prevalent on the market. In this study, a novel method for the rapid detection of adulteration in LRS based on color reaction between ferrous sulfate (FS) and (-)-gallocatechin (GC) in LRS was studied. The polyphenols in LRS were analyzed by liquid chromatography-mass spectrometry (LC-MS/MS) as GC and (+)-catechin, and GC could show a bluish color with FS solution. LRS showed a bluish color when FS solution (0.01 mol/L) was used as the chromogenic agent, which was not observed for other ten kinds of starches tested in this work. There was an obvious color gradient when FS solution was used to react with adulterated LRS at different proportions, and the change in blue color was obvious when maize starch or cassava starch was adulterated in LRS at the ratio of 6:4. Then, a semi-quantitative method was used to make a color card according to the color gradient, which was then successfully used to detect the adulteration of four kinds of commercial LRS. The results indicate that the proposed method may facilitate the simple, effective and efficient detection of adulteration in LRS.

Consumption of low levels of egg already can evoke harmful physiological responses in humans in those allergic to eggs. By detection of egg in food products, using Egg ELISA kits to determine its unintended presence, food producers can respond to avoid potential safety or quality risks of their products. Selection of an ELISA kit fit for the issue at hand is challenging due to, amongst others, lack of information on assay performances with specified matrices. In this study, performances of seven commercial egg ELISA kits are compared for nine different relevant matrices: cookie, chocolate, pasta, dressing, stock cube, wine, vegetable drink and milk, ice cream and meat/meat replacers. The presence of egg was unified for all ELISA kits to mg total egg protein kg−1 food product. In every matrix, kit performances for recovery, intra- and interassay were compared, and also processing is accounted for by determination of egg in incurred samples. All seven kits were able to detect egg qualitatively at the VITAL3 ED01 level of 0.2 mg total egg protein and the corresponding relevant portion size for each matrix. For quantitative results, each ELISA kit showed an increase in detected egg concentration with increased egg levels and performed within the set criteria for recovery for the cookie, chocolate, stock cube and wine. For pasta, vegetable drink and milk, ice cream, and salad dressing, recovery of egg was within the set criteria for at least 4 ELISA kits. Most challenging matrices were meat/meat replacers, showing high matrix effects which could not be explained by the possible egg presence in the cognate blank. Only one ELISA kit was able to recover egg within the set criteria for the meat/meat replacer matrix. Results enable food industry to choose for ELISA kits suitable for egg detection in the matrix of interest.

[Display omitted] Multi-attribute methods employing mass spectrometry are applied throughout the biopharmaceutical industry for product and process characterization purposes but are not yet widely accepted as a method for batch release and stability testing under good manufacturing practice (GMP) due to limited experience and level of comfort with the technical, compliance and regulatory aspects of its implementation at quality control (QC) laboratories. Here, current literature related to the development and application of the multi-attribute method by peptide mapping liquid chromatography mass spectrometry (MAM) is compiled with the aim of providing guidance for the implementation of MAM in a QC laboratory. This article, focusing on technical considerations, is the first part of a two-tiered publication, whereby the second part will focus on GMP compliance and regulatory aspects. This publication has been prepared by a group of industry experts representing 14 globally acting major biotechnology companies under the umbrella of the European Federation of Pharmaceutical Industries and Associations (EFPIA) Manufacturing & Quality Expert Group (MQEG). [ABSTRACT FROM AUTHOR]

Despite recent developments in managing metastatic melanomas, patients' overall survival remains low. Therefore, the current study aims to understand better the proteome‐wide changes associated with melanoma metastasis that will assist with identifying targeted therapies. The latest development in mass spectrometry‐based proteomics, together with extensive bioinformatics analysis, was used to investigate the molecular changes in 60 formalin‐fixed and paraffin‐embedded samples of primary and lymph nodes (LN) and distant organ metastatic melanomas. A total of 4631 proteins were identified, of which 72 and 453 were significantly changed between the LN and distant organ metastatic melanomas compared to the primary lesions (adj. p‐value <0.05). An increase in proteins such as SLC9A3R1, CD20 and GRB2 and a decrease in CST6, SERPINB5 and ARG1 were associated with regional LN metastasis. By contrast, increased metastatic activities in distant organ metastatic melanomas were related to higher levels of CEACAM1, MC1R, AKT1 and MMP3‐9 and decreased levels of CDKN2A, SDC1 and SDC4 proteins. Furthermore, machine learning analysis classified the lesions with up to 92% accuracy based on their metastatic status. The findings from this study provide up to date proteome‐level information about the progression of melanomas to regional LN and distant organs, leading to the identification of protein signatures with potential for clinical translation. [ABSTRACT FROM AUTHOR]

For the extraction and measurement of plasma amino acids using LC/ESI–MS/MS technology, a straightforward and dependable approach has been established. The approach employs a small amount of sample and fully quantifies it using a labeled internal standard of amino acids. Water, formic acid, and methanol were used as the mobile phase in a gradient method to obtain clear separation. On a Shimatzu mass spectrometer 8030, optimized multiple reaction monitoring (MRM) was utilized to find amino acids. All amino acids have coefficients of correlation that range from 0.91 to 0.99, and they are all linear over their respective reference ranges. Precision's intra-day and inter-day coefficients of variation (CV %) ranged from 3.29 to 11.73% and 5.04 to 12.48%, respectively. Amino acid recovery ranged from 92.1 to 108.2%. [ABSTRACT FROM AUTHOR]

In this work, we studied the potential of supercritical fluid CO2 technology to extract almond wastes and obtain a fibre product rich in minerals and phenolics without the use of an extraction co-solvent. The analysis of phenolics in the resulting extracted product was performed using liquid chromatography–tandem mass spectrometry (LC-MS/MS) and showed vanillin, catechin, and dihydroxybenzoic, vanillic, and syringic acids as main phenolic compounds (PC). In addition, the analysis of minerals carried out using inductively coupled plasma optical emission spectroscopy (ICP-OES) showed a wide range of macroelements like magnesium (Mg) and potassium (K) in quantities of up to 1.7 g/kg (Mg) and 6 g/kg (K), representing a value matrix that may be integrated into functional drinks targeting athletic people while promoting a circular economy and food up-cycling. [ABSTRACT FROM AUTHOR]

Background: Neuromyelitis optica spectrum disorder (NMOSD) is an autoimmune central nervous system (CNS) inflammatory and demyelinating disorder that can lead to serious disability and mortality. Humoral fluid biomarkers with specific, convenient, and efficient profiles that could characterize and monitor disease activity or severity are very useful. We aimed to develop a sensitive and highthroughput liquid chromatography-tandem mass spectrometry (LC-MS)/MSbased analytical method for novel biomarkers finding in NMOSD patients and verified its function tentatively. Methods: Serum samples were collected from 47 NMOSD patients, 18 patients with other neurological disorders (ONDs), and 35 healthy controls (HC). Cerebrospinal fluid (CSF) samples were collected from 18 NMOSD and 17 OND patients. Three aromatic amino acids (phenylalanine, tyrosine, and tryptophan) and nine important metabolites that included phenylacetylglutamine (PAGln), indoleacrylic acid (IA), 3-indole acetic acid (IAA), 5-hydroxyindoleacetic acid (HIAA), hippuric acid (HA), I-3-carboxylic acid (I-3-CA), kynurenine (KYN), kynurenic acid (KYNA), and quinine (QUIN) were analyzed by using the liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based method. The profile of IA was further analyzed, and its function was verified in an astrocyte injury model stimulated by NMO-IgG, which represents important events in NMOSD pathogenesis. Results: In the serum, tyrosine and some of the tryptophan metabolites IA and I-3-CA decreased, and HIAA increased significantly in NMOSD patients. The CSF levels of phenylalanine and tyrosine showed a significant increase exactly during the relapse stage, and IA in the CSF was also increased markedly during the relapse and remission phases. All conversion ratios had similar profiles with their level fluctuations. In addition, the serum IA levels negatively correlated with glial fibrillary acidic protein (GFAP), and neurofilament light (NfL) levels in the serum of NMOSD patients were measured by using ultra-sensitive single-molecule arrays (Simoa). IA showed an anti-inflammatory effect in an in vitro astrocyte injury model. Conclusion: Our data suggest that essential aromatic amino acid tryptophan metabolites IA in the serum or CSF may serve as a novel promising biomarker to monitor and predict the activity and severity of NMOSD disease. Supplying or enhancing IA function can promote anti-inflammatory responses and may have therapeutic benefits. [ABSTRACT FROM AUTHOR]

An immobilized metal affinity (IMAC) adsorbent was prepared for selective enrichment of adenine type CKs, via grafting polymer chain pendant with iminodiacetic acid (IDA) from polydopamine (PDA)/polyethyleneimine (PEI)-coated magnetic graphene oxide (magGO) via surface-initiated-atom transfer radical polymerization (SI-ATRP). The prepared IMAC sorbent exhibited remarkable adsorption performances and good selectivity for adenine-type CKs and was utilized as a sorbent of magnetic solid-phase extraction (MSPE) for effective enrichment of four adenine-type CKs in bean sprouts. Under the optimized extraction conditions, an analytical method for four adenine type CKs in bean sprouts was established by combining the MSPE combined with ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The recoveries of the analytes were between 80.4 ± 1.9% and 114.6 ± 1.5% (n = 3). The limits of detection (LODs) range from 0.63 to 2.30 pg⋅mL−1. The relative standard deviations of intra-day and inter-day were less than 12.6%. The established method was successfully applied to the selective extraction and sensitive detection of trace adenine-type CKs in plant samples. [ABSTRACT FROM AUTHOR]

Collection of finger sweat is explored here as a rapid and convenient way of monitoring patient adherence to antipsychotic drugs. Finger sweat samples (n = 426) collected from patients receiving treatment with clozapine, quetiapine and olanzapine were analysed by liquid chromatography mass spectrometry, including a subgroup of patients with paired plasma samples. Finger sweat samples were also analysed from a negative control group and patients who had handled antipsychotic medication only. The finger sweat test (based on the detection of parent drug in one donated sample) was 100% effective in monitoring adherence within commonly prescribed dosing ranges. In comparison to participants who handled the medication only, the test could distinguish between contact and administration through monitoring of the drug metabolite, or the level of parent drug. Additionally, in a subgroup of patients prescribed clozapine, a statistically significant correlation was observed between the mass of parent drug in finger sweat and plasma concentration. The finger sweat technology shows promise as a dignified, noninvasive method to monitor treatment adherence in patients taking antipsychotics.

Honey is known to have antimicrobial, immunomodulatory and wound healing properties. The biological properties of honey have been attributed to phytochemicals derived from their source plants and research has focused on identifying the bioactive phytochemicals with therapeutic potential. In this study, we determined the ability of 5 honeys from Kazakhstan and manuka honey to stimulate TNF-α and TGF-β production by human keratinocytes. TNF-α and TGF-β levels increased over time in honey treated and untreated keratinocytes, whereas cells treated with sugar solutions that matched those of the honeys had reduced levels of both cytokines. This suggests that the non-sugar phytochemical components of the honeys may have prevented this decrease. Analysis by LC-MS confirmed that the honeys contained a diverse range of phytochemicals. Some phytochemicals e.g. pinobanksin and vanillin were present at different levels across the honey types, whereas other components, e.g. dicarboxylic acids and their glycosides, were abundant in all honeys. [ABSTRACT FROM AUTHOR]

Manipulating the crop load in peach trees determines carbon supply and optimum balance between fruit yield and quality potentials. The impact of carbon supply on peach fruit quality was assessed in three development stages (S2, S3, S4) on fruit of equal maturity from trees that were carbon (C) starved (unthinned) and sufficient (thinned). Previous studies determined that primary metabolites of peach fruit mesocarp are mainly linked with developmental processes, thus, the secondary metabolite profile was assessed using non-targeted liquid chromatography mass-spectrometry (LC-MS). Carbon sufficient (C-sufficient) fruit demonstrated superior quality attributes as compared to C-starved fruit. Early metabolic shifts in the secondary metabolome appear to prime quality at harvest. Enhanced C-availability facilitated the increased and consistent synthesis of flavonoids, like catechin, epicatechin and eriodyctiol, via the phenylpropanoid pathway, providing a link between the metabolome and fruit quality, and serving as signatures of C-sufficiency during peach fruit development. [Display omitted] • Optimized crop loads enhance C-supply priming peach fruit metabolism and quality. • Primary metabolism is mainly linked with peach fruit developmental processes. • Non-targeted LC-MS revealed that flavonoids up-accumulate in the C-sufficient fruit. • Phenylpropanoids are signatures of C-sufficiency during peach fruit development. • Phenylpropanoids link primary/secondary metabolism with superior fruit quality. [ABSTRACT FROM AUTHOR]

Plants are an indispensable cornerstone of sustainable global food supply. While immense progress has been made in decoding the genomes of crops in recent decades, the composition of their proteomes, the entirety of all expressed proteins of a species, is virtually unknown. In contrast to the model plant Arabidopsis thaliana, proteomic analyses of crop plants have often been hindered by the presence of extreme concentrations of secondary metabolites such as pigments, phenolic compounds, lipids, carbohydrates or terpenes. As a consequence, crop proteomic experiments have, thus far, required individually optimized protein extraction protocols to obtain samples of acceptable quality for downstream analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS). In this article, we present a universal protein extraction protocol originally developed for gel-based experiments and combined it with an automated single-pot solid-phase-enhanced sample preparation (SP3) protocol on a liquid handling robot to prepare high-quality samples for proteomic analysis of crop plants. We also report an automated offline peptide separation protocol and optimized micro-LC-MS/MS conditions that enables the identification and quantification of ~10,000 proteins from plant tissue within 6 h of instrument time. We illustrate the utility of the workflow by analyzing the proteomes of mature tomato fruits to an unprecedented depth. The data demonstrate the robustness of the approach which we propose for use in upcoming large-scale projects that aim to map crop tissue proteomes. [ABSTRACT FROM AUTHOR]

Olanzapine is a thienobenzodiazepine class antipsychotic that strongly antagonises the 5-HT2A serotonin receptor, but acute poisonings are reported rarely. Symptoms of an overdose include disorder of consciousness, hypersalivation, myosis, and coma. Serum concentration higher than 0.1 mg/L is toxic, while concentration above 1 mg/L can be fatal. Here we report key data about 61 patients admitted to the National Poison Control Centre in Belgrade, Serbia over olanzapine poisoning in 2017 and 2018. The ingested doses ranged from 35 to 1680 mg, and time from ingestion to determination from two to 24 hours. In 34 patients olanzapine serum concentrations were in the therapeutic range and in 27 in the toxic range. In five patients they were higher than fatal, but only one patient died. The most common symptoms of poisoning were depressed consciousness (fluctuating from somnolence to coma), tachycardia, hypersalivation, hypotension, myosis, and high creatine kinase. All patients but one recovered fully after nonspecific detoxification and symptomatic and supportive therapy.

Abstract Background Takeda G protein receptor 5 (TGR5) is widely recognized as a potential drug target for the treatment of metabolic diseases. TGR5 is not only a metabolic regulator, but also has a potential role that participating in developing and progressing of gastrointestinal cancer. We aimed to investigate the potential role of TGR5 in pancreatic cancer by utilizing molecular experiments and the liquid chromatography mass spectrometry (LC–MS) based metabolomics. Methods Herein, we assessed pancreatic cancer proliferation, migration and invasion in response to TGR5 antagonist SBI-115 in vitro experiments. Cell death was examined by using TUNEL assay on agarose-embedded sections. Then we investigated the effects of TGR5 on PANC-1 and BXPC3 cells via transmission electron microscopy (TEM). Moreover, LC–MS-based metabolomics was performed to explore the potential underlying mechanisms of TGR5 in pancreatic cancer. The correlations between TGR5 and the metabolism-related genes were further analysed by GEPIA 2 database. Results We found the proliferation capacities were decreased significantly in PANC-1 and BXPC3 cells after the treatment of SBI-115 for 48 h. The results of TUNEL assay showed that antagonism of TGR5 by SBI-115 had a remarkable effect on inducing cell death. Analysis of TEM demonstrated that SBI-115 treatment could impair the morphology of mitochondria in most PANC-1 and BXPC3 cells. The LC–MS-based analyses revealed that antagonism of TGR5 could alter the metabolic profiles of PANC-1 cells in vitro. Moreover, TGR5 was associated with some metabolism-related genes in pancreatic cancer. Conclusion Our data suggests that antagonism of TGR5 may suppress cell proliferation and induce apoptosis in pancreatic cancer cells. TGR5 may affect the metabolism of pancreatic cancer, and TGR5 would be an attractive target for pancreatic cancer treatment.

• 21T FT-ICR MS molecular formula library annotated 53% of diatom exometabolome. • Formula library increased annotations 10x relative to MS/MS spectra matching. • FT-ICR MS assignments corroborated 94% of MS/MS spectra matching assignments. • Novel approach identified 11 protein-like compounds exuded in response to Fe stress. Increased accessibility of liquid chromatography mass spectrometry (LC-MS) metabolomics instrumentation and software have expanded their use in studies of dissolved organic matter (DOM) and exometabolites released by microbes. Current strategies to annotate metabolomes generally rely on matching tandem MS/MS spectra to databases of authentic standards. However, spectral matching approaches typically have low annotation rates for DOM. An alternative approach is to annotate molecular formula based on accurate mass and isotopic fine structure measurements that can be obtained from state-of-the-art ultrahigh resolution Fourier Transform Ion Cyclotron Resonance mass spectrometry (FT-ICR MS), but instrument accessibility for large metabolomic studies is generally limited. Here, we describe a strategy to annotate exometabolomes obtained from lower resolution LC-MS systems by matching metabolomic features to a molecular formula library generated for a representative sample analyzed by LC 21T- FT-ICR MS. The molecular formula library approach successfully annotated 53% of exometabolome features of the marine diatom Phaeodactylum tricornutum – a nearly ten-fold increase over the 6% annotation rate achieved using a conventional MS/MS approach. There was 94% agreement between assigned formula that were annotated with both approaches, and mass error analysis of the discrepancies suggested that the FT-ICR MS formula assignments were more reliable. Differences in the exometabolome of P. tricornutum grown under iron replete and iron limited conditions revealed 668 significant metabolites, including a suite of peptide-like molecules released by P. tricornutum in response to iron deficiency. These findings demonstrate the utility of FT-ICR MS formula libraries for extending the accuracy and comprehensiveness of metabolome annotations. [ABSTRACT FROM AUTHOR]

Introduction: Doping and steroid use represent a serious threat to animal health and can even lead to their untimely and painful death. However, doping is an acute problem in today’s animal racing world, particularly in camel racing. Testosterone and its ten esters (benzoate, valerate, isocaproate, hexahydrobenzoate, decanoate, undecanoate, laurate, enanthate, cypionate, and caproate) are of utmost importance, because when they are administered to animals it is difficult to measure them efficiently. The levels of testosterone and its esters in camels and other animals are typically determined using urine and blood tests. The aim of this study was to develop and validate a liquid chromatographic–mass spectrometric (LC-MS/MS) method to determine testosterone esters in camel hair, and to apply the validated method to determine testosterone esters in collected samples. To our knowledge, this is the first report of such research. Results and Discussion: The levels of testosterone and its ten derivatives, along with the cortisol-D4 internal standard, were optimised for LC–MS/MS analysis; however, only testosterone along with its seven esters (namely benzoate, valerate, isocaproate, hexahydrobenzoate, decanoate, undecanoate and laurate) could be validated in camel hair. Only five testosterone esters could be determined in camel hair samples; the concentrations were obtained as 10.5–14.9 pg/mg for valerate (in three camels), 12.5–151.6 pg/mg for hexahydrobenzoate (in six camels), 4.8–32.1 pg/mg for laurate (in five camels), 5.1 pg/mg decanoate (in one camel), and 8.35–169 pg/mg for testosterone (in all 24 camels). Interestingly, the three racing camels displayed high concentrations of testosterone (59.2–169 pg/mg, all three camels), laurate (4.8–14.5 pg/mg, two camels), hexahydrobenzoate (116 pg/mg, one camel), decanoate (5.1 pg/mg, one camel), and valerate (11.7 pg/mg, one camel). Methods: Camel hair samples were collected from 21 non-racing dromedary camels along with three racing camels in Al Ain, UAE; these were decontaminated, pulverised, sonicated, and extracted prior to analysis. An LC–MS/MS method was employed to determine the levels of testosterone esters in the hair samples. Conclusions: This novel camel-hair test procedure is accurate, sensitive, rapid, and robust. The findings reported in this study could be significant to evaluate racing camels for suspected doping offenses. Further controlled testosterone supplementation studies are required to evaluate individual esters’ effects on camel health and diseases and on performance enhancement levels. This new hair test could promote further studies in doping control, toxicology, and pharmacology, as well as having other clinical applications relating to camel health, injury, and disease.

Neuropathic pain is a condition affecting the quality of life of a substantial part of the population, but biomarkers and treatment options are still limited. While this type of pain is caused by nerve damage, in which lipids play key roles, lipidome alterations related to nerve injury remain poorly studied. Here, we assessed blood lipidome alterations in a common animal model, the rat sciatic nerve crush injury. We analyzed alterations in blood lipid abundances between seven rats with nerve injury (NI) and eight control (CL) rats in a time-course experiment. For these rats, abundances of 377 blood lipid species were assessed at three distinct time points: immediately after, two weeks, and five weeks post injury. Although we did not detect significant differences between NI and CL at the first two time points, 106 lipids were significantly altered in NI five weeks post injury. At this time point, we found increased levels of triglycerides (TGs) and lipids containing esterified palmitic acid (16:0) in the blood plasma of NI animals. Lipids containing arachidonic acid (20:4), by contrast, were significantly decreased after injury, aligning with the crucial role of arachidonic acid reported for NI. Taken together, these results indicate delayed systematic alterations in fatty acid metabolism after nerve injury, potentially reflecting nerve tissue restoration dynamics. [ABSTRACT FROM AUTHOR]

Sphingolipids (SLs) are essential lipids with important functions in membrane formation and cell signaling. The presence of a long chain base (LCB) structure is common to all SLs. De novo SL synthesis is initiated by the enzyme serine-palmitoyltransferase (SPT), which forms an LCB by the conjugation from serine and fatty acyl-CoAs. SPT can metabolize a variety of acyl-CoA substrates, which form diverse LCB structures within and across species. The LCB then undergoes further metabolic modifications resulting in an extraordinarily diverse spectrum of sphingolipids formed. SL analysis, using liquid chromatography-mass spectrometry (LC-MS)-based methods, poses challenges due to the diverse range of frequently isobaric species. This complexity complicates the identification of underlying LCB structures using standard lipidomics approaches. Here, we describe a simplified method to analyze the LCB profile in cells, tissue, and blood. The procedure involves chemical hydrolysis to remove the conjugated headgroups and N-acyl chains, allowing to specifically resolve the underlying LCB structures by LC-MS. This method can also be combined with an isotope labeling approach to determine in vivo SPT activity and total SL de novo synthesis over time., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Studies suggest that short chain fatty acids (SCFAs), which are primarily produced from fermentation of fiber, regulate insulin secretion through free fatty acid receptors 2 and 3 (FFA2 and FFA3). As these are G-protein coupled receptors (GPCRs), they have potential therapeutic value as targets for treating type 2 diabetes (T2D). The exact mechanism by which these receptors regulate insulin secretion and other aspects of pancreatic β cell function is unclear. It has been reported that glucose-dependent release of acetate from pancreatic β cells negatively regulates glucose stimulated insulin secretion. While these data raise the possibility of acetate's potential autocrine action on these receptors, these findings have not been independently confirmed, and multiple concerns exist with this observation, particularly the lack of specificity and precision of the acetate detection methodology used., Methods: Using Min6 cells and mouse islets, we assessed acetate and pyruvate production and secretion in response to different glucose concentrations, via liquid chromatography mass spectrometry., Results: Using Min6 cells and mouse islets, we showed that both intracellular pyruvate and acetate increased with high glucose conditions; however, intracellular acetate level increased only slightly and exclusively in Min6 cells but not in the islets. Further, extracellular acetate levels were not affected by the concentration of glucose in the incubation medium of either Min6 cells or islets., Conclusions: Our findings do not substantiate the glucose-dependent release of acetate from pancreatic β cells, and therefore, invalidate the possibility of an autocrine inhibitory effect on glucose stimulated insulin secretion.

Oxysterols and bile acids are interconnected bioactive lipids playing pivotal roles in diverse physiological and pathological processes. For this reason, they are increasingly studied together for their implications in various diseases. However, due to analytical challenges inherent to the nature of these analytes, very few methods have been developed for the simultaneous analysis of these lipids. We here report the development of a sensitive LC-MS/MS method for the combined quantification of 18 oxysterols, 11 unconjugated, 15 conjugated bile acids, and 1 bile acid precursor, using 8 isotope-labeled internal standards, addressing the need for a more comprehensive analysis of these interesting lipid families. During the method development, we investigated different extraction protocols, set up a purification step, and achieved chromatographic separation for these lipids, overcoming challenges such as the large number of analytes, isomers, and wide range of polarity across the analytes. Finally, the method was successfully applied to the analysis of preclinical and clinical samples, quantifying 12 oxysterols and 14 bile acids in human plasma, 10 oxysterols and 18 bile acids in mouse plasma from the vena cava, and 10 oxysterols and 20 bile acids in mouse plasma from the portal vein within a single chromatographic run., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Filipendula ulmaria, commonly known as meadowsweet, is a wild herbaceous flowering plant that is widely distributed in Europe. A range of salicylic acid derivatives and flavonol glycosides have been previously associated with the antirheumatic and diuretic properties of F. ulmaria. In the present work, a hydroalcoholic extract from F. ulmaria aerial parts was extensively profiled using an efficient NMR-based dereplication strategy. The approach involves the fractionation of the crude extract by centrifugal partition chromatography (CPC), 13C NMR analysis of the fractions, 2D-cluster mapping of the entire NMR dataset, and, finally, structure elucidation using a natural metabolite database, validated by 2D NMR data interpretation and liquid chromatography coupled with mass spectrometry. The chemodiversity of the aerial parts was extensive, with 28 compounds unambiguously identified, spanning various biosynthetic classes. The F. ulmaria extract and CPC fractions were screened for their potential to enhance skin epidermal barrier function and skin renewal properties using in vitro assays performed on Normal Human Epidermal Keratinocytes. Fractions containing quercetin, kaempferol glycosides, ursolic acid, pomolic acid, naringenin, β-sitosterol, and Tellimagrandins I and II were found to upregulate genes related to skin barrier function, epidermal renewal, and stress responses. This research is significant as it could provide a natural solution for improving hydration and skin renewal properties.

Hepatic AL amyloidosis patients have hepatomegaly and elevated alkaline phosphatase levels; and gastrointestinal AL amyloidosis presents with dysphagia or loss of appetite. AL amyloidosis, also known as primary systemic amyloidosis, is a rare disease with an estimated incidence of 9.7 to 14.0 cases per million person years.1 Ninety-five percent of AL amyloidosis patients are >40 years old with a median age of 64 to 70 years old; 65% to 70% of patients are male. Keywords: amyloid; light chain; liquid chromatography mass spectrometry EN amyloid light chain liquid chromatography mass spectrometry 889 892 4 09/21/22 20221001 NES 221001 INTRODUCTION The three main types of systemic amyloidosis are: primary amyloid light-chain (AL), secondary, and familial amyloidosis. [Extracted from the article]