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Glucagon plays a central role in amino acid (AA) homeostasis. The dog is an established model of glucagon biology, and recently, metabolomic changes in people associated with glucagon infusions have been reported. Glucagon also has effects on the kidney; however, changes in urinary AA concentrations associated with glucagon remain under investigation. Therefore, we aimed to fill these gaps in the canine model by determining the effects of glucagon on the canine plasma metabolome and measuring urine AA concentrations. Employing two constant rate glucagon infusions (CRI) - low-dose (CRI-LO: 3 ng/kg/min) and high-dose (CRI-HI: 50 ng/kg/min) on five research beagles, we monitored interstitial glucose and conducted untargeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) on plasma samples and urine AA concentrations collected pre- and post-infusion. The CRI-HI induced a transient glucose peak (90-120 min), returning near baseline by infusion end, while only the CRI-LO resulted in 372 significantly altered plasma metabolites, primarily reductions (333). Similarly, CRI-HI affected 414 metabolites, with 369 reductions, evidenced by distinct clustering post-infusion via data reduction (PCA and sPLS-DA). CRI-HI notably decreased circulating AA levels, impacting various AA-related and energy-generating metabolic pathways. Urine analysis revealed increased 3-methyl-l-histidine and glutamine, and decreased alanine concentrations post-infusion. These findings demonstrate glucagons glucose-independent modulation of the canine plasma metabolome and highlight the dogs relevance as a translational model for glucagon biology. Understanding these effects contributes to managing dysregulated glucagon conditions and informs treatments impacting glucagon homeostasis.

There is great concern in equine sport over the potential use of pharmaceutical agents capable of editing the genome or modifying the expression of gene products. Synthetic oligonucleotides are short, single-stranded polynucleotides that represent a class of agents capable of modifying gene expression products with a high potential for abuse in horseracing. As these substances are not covered by most routine anti-doping analytical approaches, they represent an entire class of compounds that are not readily detectable. The nucleotide sequence for each oligonucleotide is highly specific, which makes targeted analysis for these agents problematic. Accordingly, we have developed a non-targeted approach to detect the presence of specific product ions that are not naturally present in ribonucleic acids. Briefly, serum samples were extracted using solid-phase extraction with a mixed-mode cartridge following the disruption of protein interactions to isolate the oligonucleotides. Following the elution and concentration steps, chromatographic separation was achieved utilizing reversed-phase liquid chromatography. Following an introduction to a Thermo Q Exactive HF mass spectrometer using electrospray ionization, analytes were detected utilizing a combination of full-scan, parallel reaction monitoring and all ion fragmentation scan modes. The limits of detection were determined along with the accuracy, precision, stability, recovery, and matrix effects using a representative 13mer oligonucleotide. Following method optimization using the 13mer oligonucleotide, the method was applied to successfully detect the presence of specific product ions in three unique oligonucleotide sequences targeting equine-specific transcripts.

Monofluoroacetate is a highly lethal toxin that causes death by inhibiting cellular adenosine triphosphate (ATP) production. The heart and brain are the primary target organs. Acute death is attributed to cardiac fibrillation and/or convulsions. Although it occurs naturally in some plants, a major source of animal intoxication is access to sodium monofluoroacetate (NaMFA) pesticide, which continues to be a concern in the USA and around the world despite restricted use in some countries including the USA. There are also concerns about misuse of this pesticide for malicious poisoning. Currently, a tissue-based diagnostic method for NaMFA intoxication in animals is lacking. There is a critical need by the veterinary diagnostic community for a simple, sensitive and reliable tissue-based diagnostic test to confirm NaMFA poisoning in animals. We have developed and extensively evaluated a sensitive novel liquid chromatography combined with tandem mass spectrometry method suitable for this purpose. The limits of detection and limits of quantitation are 1.7 and 5.0 ng/g, respectively. The accuracy and precision met or exceeded expectations. The method performance was verified using the incurred kidney obtained from animal diagnostic cases. This novel kidney-based method is now available for clinical use and can help with diagnostic purposes, including detecting potential issues related to animal foods.

Protein lipidation dynamically controls protein localization and function within cellular membranes. A unique form of protein O-fatty acylation in Corynebacterium, termed protein O-mycoloylation, involves the attachment of mycolic acids─unusually large and hydrophobic fatty acids─to serine residues of proteins in these organisms outer mycomembrane. However, as with other forms of protein lipidation, the scope and functional consequences of protein O-mycoloylation are challenging to investigate due to the inherent difficulties of enriching and analyzing lipidated peptides. To facilitate the analysis of protein lipidation and enable the comprehensive profiling and site mapping of protein O-mycoloylation, we developed a chemical proteomics strategy integrating metabolic labeling, click chemistry, cleavable linkers, and a novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method employing LC separation and complementary fragmentation methods tailored to the analysis of lipophilic, MS-labile O-acylated peptides. Using these tools in the model organism Corynebacterium glutamicum, we identified approximately 30 candidate O-mycoloylated proteins, including porins, mycoloyltransferases, secreted hydrolases, and other proteins with cell envelope-related functions─consistent with a role for O-mycoloylation in targeting proteins to the mycomembrane. Site mapping revealed that many of the proteins contained multiple spatially proximal modification sites, which occurred predominantly at serine residues surrounded by conformationally flexible peptide motifs. Overall, this study (i) discloses the putative protein O-mycoloylome for the first time, (ii) yields new insights into the undercharacterized proteome of the mycomembrane, which is a hallmark of important pathogens (e.g., Corynebacterium diphtheriae, Mycobacterium tuberculosis), and (iii) provides generally applicable chemical strategies for the proteomic analysis of protein lipidation.

Sphingolipids are an essential subset of bioactive lipids found in most eukaryotic cells that contribute to membrane biophysical properties and are involved in cellular differentiation, recognition, and mediating interactions. The described nanoHPLC-ESI-Q/ToF methodology utilizes known biosynthetic pathways, accurate mass detection, optimized collision-induced disassociation, and a robust nanoflow chromatographic separation for the analysis of intact sphingolipids found in human tissue, cells, and serum. The methodology was developed and validated with an emphasis on addressing the common issues experienced in profiling these amphipathic lipids, which are part of the glycocalyx and lipidome. The high sensitivity obtained using nanorange flow rates with robust chromatographic reproducibility over a wide range of concentrations and injection volumes results in confident identifications for profiling these low-abundant biomolecules.

Isomerized amino acid residues have been identified in many peptides extracted from tissues or excretions of humans and animals. These isomerized residues can play key roles by affecting biological activity or by exerting an influence on the process of aging. Isomerization occurs spontaneously and does not result in a mass shift. Thus, identifying and localizing isomerized residues in biological samples is challenging. Herein, we introduce a fast and efficient method using tandem mass spectrometry (MS) to locate isomerized residues in peptides. Although MS2 spectra are useful for identifying peptides that contain an isomerized residue, they cannot reliably localize isomerization sites. We show that this limitation can be overcome by utilizing MS3 experiments to further evaluate each fragment ion from the MS2 stage. Comparison at the MS3 level, utilizing statistical analyses, reveals which MS2 fragments differ between samples and, therefore, must contain the isomerized sites. The approach is similar to previous work relying on ion mobility to discriminate MS2 product ions by collision cross-section. The MS3 approach can be implemented using either ion-trap or beam-type collisional activation and is compatible with the quantification of isomer mixtures when coupled to a calibration curve. The method can also be implemented in combination with liquid chromatography in a targeted approach. Enabling the identification and localization of isomerized residues in peptides with an MS-only methodology will expand accessibility to this important information.

Liquid chromatography with mass spectrometry (LC-MS)-based metabolomics detects thousands of molecular features (retention time-m/z pairs) in biological samples per analysis, yet the metabolite annotation rate remains low, with 90% of signals classified as unknowns. To enhance the metabolite annotation rates, researchers employ tandem mass spectral libraries and challenging in silico fragmentation software. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) may offer an additional layer of structural information in untargeted metabolomics, especially for identifying specific unidentified metabolites that are revealed to be statistically significant. Here, we investigate the potential of hydrophilic interaction liquid chromatography (HILIC)-HDX-MS in untargeted metabolomics. Specifically, we evaluate the effectiveness of two approaches using hypothetical targets: the post-column addition of deuterium oxide (D2O) and the on-column HILIC-HDX-MS method. To illustrate the practical application of HILIC-HDX-MS, we apply this methodology using the in silico fragmentation software MS-FINDER to an unknown compound detected in various biological samples, including plasma, serum, tissues, and feces during HILIC-MS profiling, subsequently identified as N1-acetylspermidine.

Short-chain fatty acids (SCFAs) comprise the largest group of gut microbial fermentation products. While absorption of most nutrients occurs in the small intestine, indigestible dietary components, such as fiber, reach the colon and are processed by the gut microbiome to produce a wide array of metabolites that influence host physiology. Numerous studies have implicated SCFAs as key modulators of host health, such as in regulating irritable bowel syndrome (IBS). However, robust methods are still required for their detection and quantitation to meet the demands of biological studies probing the complex interplay of the gut-host-health paradigm. In this study, a sensitive, rapid-throughput, and readily expandible UHPLC-QqQ-MS platform using 2-PA derivatization was developed for the quantitation of gut-microbially derived SCFAs, related metabolites, and isotopically labeled homologues. The utility of this platform was then demonstrated by investigating the production of SCFAs in cecal contents from mice feeding studies, human fecal bioreactors, and fecal/bacterial fermentations of isotopically labeled dietary carbohydrates. Overall, the workflow proposed in this study serves as an invaluable tool for the rapidly expanding gut-microbiome and precision nutrition research field.

BACKGROUND: A better understanding of treatment progression and recovery in pulmonary tuberculosis (TB) infectious disease is crucial. This study analyzed longitudinal serum samples from pulmonary TB patients undergoing interventional treatment to identify surrogate markers for TB-related outcomes. METHODS: Serum that was collected at baseline and 8, 17, 26, and 52 weeks from 30 TB patients experiencing durable cure were evaluated and compared using a sensitive LC-MS/MS proteomic platform for the detection and quantification of differential host protein signatures relative to timepoint. The global proteome signature was analyzed for statistical differences across the time course and between disease severity and treatment groups. RESULTS: A total of 676 proteins showed differential expression in the serum over these timepoints relative to baseline. Comparisons to understand serum protein dynamics at 8 weeks, treatment endpoints at 17 and 26 weeks, and post-treatment at 52 weeks were performed. The largest protein abundance changes were observed at 8 weeks as the initial effects of antibiotic treatment strongly impacted inflammatory and immune modulated responses. However, the largest number of proteome changes was observed at the end of treatment time points 17 and 26 weeks respectively. Post-treatment 52-week results showed an abatement of differential proteome signatures from end of treatment, though interestingly those proteins uniquely significant at post-treatment were almost exclusively downregulated. Patients were additionally stratified based upon disease severity and compared across all timepoints, identifying 461 discriminating proteome signatures. These proteome signatures collapsed into discrete expression profiles with distinct pathways across immune activation and signaling, hemostasis, and metabolism annotations. Insulin-like growth factor (IGF) and Integrin signaling maintained a severity signature through 52 weeks, implying an intrinsic disease severity signature well into the post-treatment timeframe. CONCLUSION: Previous proteome studies have primarily focused on the 8-week timepoint in relation to culture conversion status. While this study confirms previous observations, it also highlights some differences. The inclusion of additional end of treatment and post-treatment time points offers a more comprehensive assessment of treatment progression within the serum proteome. Examining the expression dynamics at these later time periods will help in the investigation of relapse patients and has provided indicative markers of response and recovery.

This paper presents the detailed design configuration and investigation of a small-scale dual-band metamaterial absorber (MTMA) for solid and liquid sensing applications. The overall dimension of the MTMA unit cell is 10 × 10 × 1.57 mm3 and constitutes an affordable FR-4 substrate. The absorber exhibits dual absorption peaks at 3.470 GHz for the S-band and 7.219 GHz for the C-band, respectively. Both absorption characteristics have been validated through comprehensive simulation and experimental procedures. The dual-band absorption rate exceeded 99% during simulations, and experimental validation showed an absorption rate above 98%. For sensing applications, various solid materials, including different Rogers substrates (RT 5880, RT 5870, RT 4003 and RT 4835) and liquids such as sunflower and crown oil, were utilized. Our findings indicate that the proposed MTMA achieves a maximum Q-factor of 191 and a sensitivity of up to 2.5 for both solid and liquid sensing applications compared to previous studies. The simulation and experimental validation of the result indicate that suggested MTMA can be effectively used in different sensing applications such as the medical and communications industry. [ABSTRACT FROM AUTHOR]

Introduction: Self-reports of electronic cigarette (ECIG) device and liquid characteristics are not always accurate or consistent with characteristics as measured by researchers. Two methods for measuring ECIG characteristics were compared: user self-reports and rater-coded pictures. Methods: Exclusive ECIG users (N = 321) reported on device (disposable, refillable, adjustable power, brand) and liquid (nicotine concentration, formulation, flavor) characteristics. To measure device type, they chose the term that best described their device ("cig-alike," "vape pen," "mod," "pod," "don't know") and the picture that best resembled their device (cig-alike, vape pen, box mod, USB-shaped pod, teardrop-shaped pod, none). Respondents uploaded device and liquid pictures, and independent raters coded these same features. Agreement between methods was examined with Cohen's kappa and intra-class correlations, including with "don't know" responses included and excluded from analyses. Results: Regardless of how "don't know" responses were treated, agreement was highest for disposable (95.3–97.7%), refillable (96.3%), adjustable power (83.6–88.7%), and brand (77.9–80.4%), and lower for nicotine concentration (72.7%), nicotine formulation (58.6–79.4%), and flavor (66.2%). For device type, agreement was moderate using both term-based (67.9–78.8%) and picture resemblance-based (71.7%) items. For terms, the greatest discrepancy was for devices classified as "vape pens" by self-reports; of these, 70.6% were classified as "pods" by raters. For picture resemblance, ∼13% of users reported that their device resembled none of the pictures; raters classified these devices as USB-shaped pods (50.0%) and mods (23.8%). Conclusions: Self-reports may be sufficient for measuring some characteristics (brand, disposable, refillable, adjustable power), but not others (nicotine concentration and formulation, and some flavor). [ABSTRACT FROM AUTHOR]

Purpose: This study was designed to conduct pooled comparisons of the relative clinical efficacy and safety of computed tomography (CT)-guided localization for pulmonary nodules (PNs) using either coil- or liquid material-based approaches. Material and methods: Relevant articles published as of July 2023 were identified in the Web of Science, PubMed, and Wanfang databases, and pooled analyses of relevant endpoints were then conducted. Results: Six articles that enrolled 287 patients (341 PNs) and 247 patients (301 PNs) that had respectively undergone CT-guided localization procedures using coil- and liquid material-based approaches prior to video-assisted thoracic surgery (VATS) were included in this meta-analysis. The liquid material group exhibited a significantly higher pooled successful localization rate as compared to the coil group (p = 0.01), together with significantly lower pooled total complication rates (p = 0.0008) and pneumothorax rates (p = 0.01). Both groups exhibited similar rates of pulmonary hemorrhage (p = 0.44) and successful wedge resection (p = 0.26). Liquid-based localization was also associated with significant reductions in pooled localization and VATS procedure durations (p = 0.004 and 0.007). Conclusions: These data are consistent with CT-guided localization procedures performed using liquid materials being safer and more efficacious than coil-based localization in patients with PNs prior to VATS resection. [ABSTRACT FROM AUTHOR]

Phononic crystal (PnC) sensors are recognized for their capability to control acoustic wave propagation through periodic structures, presenting considerable potential across various applications. Despite advancements, the effects of fluid viscosity on PnC performance remain intricate and inadequately understood. This study theoretically investigates the influence of shear (dynamic) and bulk viscosity on acoustic wave damping in defective one-dimensional phononic crystal (1D PnC) sensors designed for detecting liquid analytes. Acetic acid with varying viscosities is considered to fill a cavity layer intermediated by a multilayer stack of lead and epoxy. The effects of dynamic and bulk viscosity on the resonance characteristics of the defective mode were analyzed. Numerical results reveal that increased dynamic viscosity leads to substantial broadening and decreased intensity of resonance peaks, accompanied by a shift to higher frequencies due to enhanced elastic wave attenuation and damping. At low dynamic viscosity (η = 0.2 ηd), numerous resonance peaks with varying intensities are observed. However, at higher viscosities (η = 2.0 ηd to η = 10.0 ηd), only one prominent peak appears in the spectrum. The intensity of this resonant peak starts at 98% for η = 2 ηd and decreases to 58.8% as the dynamic viscosity increases to η = 10 ηd. Additionally, the combined effect of dynamic and bulk viscosity introduces further damping, causing a strong shift of the resonance peak to higher frequencies, along with an increase in the full width at half maximum (FWHM) and a decrease in the quality factor (QF). These findings emphasize the necessity of incorporating both shear and bulk viscosity in the design of PnC sensors to enhance their sensitivity and accuracy in practical applications. This theoretical framework provides critical insights for optimizing sensor performance and bridging gaps between theoretical predictions and experimental observations, especially in 1D PnCs, offering potential solutions to challenges in real-world PnC sensor applications. [ABSTRACT FROM AUTHOR]

The liquid is discharged into the surrounding atmospheric environment through the nozzle or orifice with small inner diameter. When the outlet pressure or flow is increased, the flow mode of the liquid at the end of the capillary can change from periodic dripping (PD) to dripping faucet (DF) with latter jetting (J). Through changing the flow rate, three typical atomization modes of PD, DF and J for deionized water and ethanol were recorded by high-speed digital camera, and the critical Weber number of the transition of atomization mode was obtained. The results show that in the process of PD, the shape of droplets is gradually changed from spherical to pear-shape with the increasing of outer diameter of the orifice. As the flowrate is increased, two liquids are gradually reached the jetting regime from the dripping mode through the dripping faucet. Compared with deionized water, the transition of ethanol exists in small range and only in the inner diameter of 0.33-0.60 mm, and it is very easy to transition directly from periodic dripping to jetting regime. The difference of atomization mode transition between deionized water and absolute ethanol is caused by the different surface tension. The critical Weber number corresponding to the transition from dripping to jetting mode can realize the prediction of the atomization mode transition process. [ABSTRACT FROM AUTHOR]

Simple Summary: Vitamins A and E are crucial for animal health, contributing to key functions such as vision, immune response, growth, reproduction, and neurological development. A deficiency in these micronutrients can lead to serious health issues. Supplementation, especially with innovative liquid formulations, is important for addressing these deficiencies and enhancing animal health and productivity. Administering these vitamins through drinking water offers a convenient method, especially during times of stress or increased nutritional demands. Injectable forms can enhance reproductive health, growth, and immune function in livestock, while liquid drops provide a precise and easy way to dose companion animals. Future research may focus on optimizing these formulations, developing specialized treatments, and leveraging nutrigenomics to customize animal nutrition, leading to advancements in veterinary care and livestock management. Vitamins A and E are vital fat-soluble micronutrients with distinct yet intertwined roles in various biological processes. This review delves into their functions, nutritional requirements across different animal species, the consequences of deficiencies, and the impact of liquid formulations on veterinary medicine and livestock production. Vitamin A exists in multiple forms, essential for vision, immunity, and growth, while vitamin E acts primarily as an antioxidant, safeguarding cell membranes from oxidative damage. Hypovitaminosis in these vitamins can lead to severe health consequences, affecting vision, immunity, growth, reproduction, and neurological functions. Hence, supplementation, particularly through innovative liquid formulations, becomes pivotal in addressing deficiencies and enhancing overall animal health and productivity. Injectable forms of vitamins A and E show promise in enhancing reproductive performance, growth, and immune function in livestock. Administering these vitamins through drinking water offers a convenient way to enhance livestock health and productivity, particularly during times of stress or increased nutritional needs. Liquid vitamin A and E drops offer a flexible and effective solution in veterinary practice, allowing precise dosing and easy administration, particularly for companion animals. Future research may aim to optimize formulations and explore targeted therapies and precision feeding via nutrigenomics, promising advancements in veterinary medicine and livestock production. [ABSTRACT FROM AUTHOR]

This study has aimed to determine the potential use of catechins derived from gambier as an anti-microbial raw material in the manufacture of hand washing liquid soap. The study started from the purification process of catechins from gambier and continued with the addition of gambier to the soap formula looking for hand washing. In this study, there were some difficulties in obtaining catechin extracts that could dissolve in water due to the character of catechins that did not dissolve completely in water. Therefore, the optimization of the catechin extraction process is optimized first. The catechin extraction process was carried out in two ways, namely by using a homogenizer at a speed of 15000 rpm for 30 minutes and a reflux process with hot water (temperature 70 degrees Celsius) for one hour. It was found that the concentration of catechin dissolved in water both in the homogenizer and reflux results was relatively the same, therefore for the manufacture of hand washing liquid soap, the process with the reflux technique would be recommended because of the cheaper operational costs.

Abstract Phononic crystal (PnC) sensors are recognized for their capability to control acoustic wave propagation through periodic structures, presenting considerable potential across various applications. Despite advancements, the effects of fluid viscosity on PnC performance remain intricate and inadequately understood. This study theoretically investigates the influence of shear (dynamic) and bulk viscosity on acoustic wave damping in defective one-dimensional phononic crystal (1D PnC) sensors designed for detecting liquid analytes. Acetic acid with varying viscosities is considered to fill a cavity layer intermediated by a multilayer stack of lead and epoxy. The effects of dynamic and bulk viscosity on the resonance characteristics of the defective mode were analyzed. Numerical results reveal that increased dynamic viscosity leads to substantial broadening and decreased intensity of resonance peaks, accompanied by a shift to higher frequencies due to enhanced elastic wave attenuation and damping. At low dynamic viscosity (η = 0.2 ηd), numerous resonance peaks with varying intensities are observed. However, at higher viscosities (η = 2.0 ηd to η = 10.0 ηd), only one prominent peak appears in the spectrum. The intensity of this resonant peak starts at 98% for η = 2 ηd and decreases to 58.8% as the dynamic viscosity increases to η = 10 ηd. Additionally, the combined effect of dynamic and bulk viscosity introduces further damping, causing a strong shift of the resonance peak to higher frequencies, along with an increase in the full width at half maximum (FWHM) and a decrease in the quality factor (QF). These findings emphasize the necessity of incorporating both shear and bulk viscosity in the design of PnC sensors to enhance their sensitivity and accuracy in practical applications. This theoretical framework provides critical insights for optimizing sensor performance and bridging gaps between theoretical predictions and experimental observations, especially in 1D PnCs, offering potential solutions to challenges in real-world PnC sensor applications.

Introduction. The essence of the acoustic – cavitation processes is that the liquid is passed through sound with a pressure at the wave surface of more than 3 bar that causes local breaks of the liquid in the vacuum phase of the wave and the collapse in the manometric phase. The opposite walls of each cavern in the collapse approach at a speed exceeding two speed of sound, due to which a high energy density is achieved at the meeting point, and what is especially valuable is the mutual transitions of energies from one form to another, unattainable under normal conditions, and, moreover, as inside cavitation area and near it. The novelty of the work is confirmed by the results of a periodic information and patent analysis, and by four patents received for inventions on the topic under consideration. Aim of the Study. The study is aimed at improving the acoustic-cavitation qualities of a disk-shaped vortex chamber used as a liquid whistle. Materials and Methods. In the study, there were used numerical modeling of flows in the FlowVision program, experimental determination of flow rates using a pitot tube, film method, removal of frequency response using SpectraPLUS 5.0, and visualization of flows and processes on optically transparent devices by the method of color indicators in stroboscopic lighting high-speed video shooting. Results. The mechanism of sound generation and noise in the flow transiting through the device has been found. The corrective effect of pump pulsations f = 300 Hz on the sound generation mechanism was revealed. The disc-shaped character of the device, which encloses the input flow in cross section from three directions, contributes to creating a more expressive acoustic signal, forming two conjugate torus vortices along the shell that ensures uniformity of the circumferential flow, attenuation of longitudinal high-frequency oscillations f = 200 kHz, and the creation of periodic zones of increased pressure along the shell. The concentrated tangential entrance to the device determines the central asymmetry of the flows in it and a number of processes that create acoustic noise. Discussion and Conclusion. The frequency of the useful acoustic signal in the vortex chamber is proportional to the speed of the transit flow, and the amplitude is proportional to the dimensions of the device. Along with the useful signal created by the interaction of the peripheral and input parts of the transit flow, noise of similar frequencies is created in the device. Other sources of noise generation are due to the presence of a concentrated tangential input. The formation of two conjugate torus vortices along the shell can be used as a means of controlling the process of interaction between parts of the transit flow. The disc-shaped vortex chamber combines the functions of sound generation and the ability to create a centrifugal field, which expands its technological capabilities.

Epidemiologic studies of low-frequency exposures or outcomes using metabolomics analyses of neonatal dried blood spots (DBS) often require assembly of samples with substantial differences in duration of storage. Independent assessment of stability of metabolites in archived DBS will enable improved design and interpretation of epidemiologic research utilizing DBS. Neonatal DBS routinely collected and stored as part of the California Genetic Disease Screening Program between 1983 and 2011 were used. The study population included 899 children without cancer before age 6 years, born in California. High-resolution metabolomics with liquid-chromatography mass spectrometry was performed, and the relative ion intensities of common metabolites and selected xenobiotic metabolites of nicotine (cotinine and hydroxycotinine) were evaluated. In total, we detected 26,235 mass spectral features across 2 separate chromatography methods (C18 hydrophobic reversed-phase chromatography and hydrophilic-interaction liquid chromatography). For most of the 39 metabolites related to nutrition and health status, we found no statistically significant annual trends across the years of storage. Nicotine metabolites were captured in the DBS with relatively stable intensities. This study supports the usefulness of DBS stored long-term for epidemiologic studies of the metabolome. -Omics-based information gained from DBS may also provide a valuable tool for assessing prenatal environmental exposures in child health research.

Four alpine goats developed diarrhea soon after the owner placed plant clippings believed to be yellow oleander (Thevetia peruviana) into their pen on a suburban property near Palm Desert, CA, USA. A 1-y-old female goat died suddenly ~1 h after eating the plant clippings and was submitted to the San Bernardino Branch of the California Animal Health and Food Safety Laboratory System for postmortem examination. The main autopsy and histopathologic findings were myocardial hemorrhage and necrosis, consistent with cardiac glycoside intoxication. Rumen contents were analyzed by LC-MS/MS; peruvoside, a cardiac glycoside, was detected, but oleandrin, the cardiac glycoside of common oleander (Nerium oleander), was not. An LC-high-resolution MS (LC-HRMS) analysis revealed the presence of peruvoside and neriifolin in the rumen contents and in a tested plant fragment, indicating that the plant was a member of the Thevetia genus. A clipping from the plant fed to the goats and submitted by the owner was identified as yellow oleander, Thevetia peruviana (also known as Cascabela thevetia).

Camponotus floridanus ants show altered behaviors followed by a fatal summiting phenotype when infected with manipulating Ophiocordyceps camponoti-floridani fungi. Host summiting as a strategy to increase transmission is also observed with parasite taxa beyond fungi, including aquatic and terrestrial helminths and baculoviruses. The drastic phenotypic changes can sometimes reflect significant molecular changes in gene expression and metabolite concentrations measured in manipulated hosts. Nevertheless, the underlying mechanisms still need to be fully characterized. To investigate the small molecules producing summiting behavior, we infected C. floridanus ants with O. camponoti-floridani and sampled their heads for LC-MS/MS when we observed the characteristic summiting phenotype. We link this metabolomic data with our previous genomic and transcriptomic data to propose mechanisms that underlie manipulated summiting behavior in "zombie ants." This "multiomic" evidence points toward the dysregulation of neurotransmitter levels and neuronal signaling. We propose that these processes are altered during infection and manipulation based on (1) differential expression of neurotransmitter synthesis and receptor genes, (2) altered abundance of metabolites and neurotransmitters (or their precursors) with known behavioral effects in ants and other insects, and (3) possible suppression of a connected immunity pathway. We additionally report signals for metabolic activity during manipulation related to primary metabolism, detoxification, and anti-stress protectants. Taken together, these findings suggest that host manipulation is likely a multi-faceted phenomenon, with key processes changing at multiple levels of molecular organization.

The use of high temperatures in conventional liquid arenga sugar processing can trigger the Maillard reaction, which causes some of the nutritional content of liquid arenga sugar to decrease. Therefore, liquid arenga sugar processing needs to be processed below 100 °C, using the vacuum evaporator process. The evaporation rate in the previously developed liquid arenga sugar processing technology is still lower than the conventional technology, which is due to the unoptimized temperature and vacuum pressure settings. In this study, a number of experiments were carried out with the aim of investigating the vacuum pressure and temperature that has the optimum evaporation rate. By utilizing the venturi effect, it can maintain vacuum conditions during the experiment. Data acquisition starts when the sample is boiling and finishes when the sample mass remains at 20%. The temperature variants comprise 60, 70, and 80 °C. While the vacuum pressure variants were 0.4 bar(-0.6 gauge), 0.3 bar(-0.7 gauge), and 0.2 bar(-0.8 gauge). Based on the conducted experiments, at a vacuum pressure of 0.3 bar(-0.7 gauge), the temperature variable dominantly affects the evaporation rate. Variant C3, at a temperature of 80 °C and a vacuum pressure of 0.2 bar(-0.8 gauge), showed better performance than the other variants. The peak value of mass loss of 7.18 kg and the peak value of mass reduction percentage of 73.12% represented it. Based on the vacuum pressure and temperature conditions of variant C3, it is estimated that processing 10 kg of liquid arenga sugar takes 2 h and 17 min, there was a reduction of almost 80% from the previous process which took 9 h 20 min.

The study of plasma generation in liquids is relevant for many applications, especially for increasing the efficiency of terahertz radiation generation. This work investigates the relationship between the laser excitation wavelength and the plasma electron density in liquid water in the near-infrared spectral range. Using numerical simulation methods based on the Keldysh theory, patterns of changes in the ionization rate and changes in the plasma electron density depending on the excitation wavelength are analyzed. The results show the mutual influence of above-threshold ionization and tunneling effects when the Keldysh parameter is close to one. A decrease in plasma electron density with increasing excitation wavelength has been shown. However, in certain wavelength ranges a local increase in plasma electron density was observed. The theoretical results obtained are consistent with the experimental data of other scientific groups. This theoretical study provides valuable information on the modulation of plasma electron density by changing laser excitation wavelengths, which is important for increasing the efficiency of terahertz radiation generation.

Hybrid halide perovskites (HHPs) are very promising absorber materials for solar cells due to their high power conversion efficiency and the low-cost solution-based processing methods. We applied small angle X-ray scattering to MAPbI3, FAPbI3 and MAPbBr3 precursor solutions in different solvents (GBL, DMF, and mixtures) to gain a deeper understanding of the building blocks during the early stage of HHP formation. We present a core–shell model where the core is formed by [PbX6] octahedra surrounded by a shell of solvent molecules, which explains the arrangement of the precursors in solution and how the solvent and the halide influence such arrangement. [ABSTRACT FROM AUTHOR]

Bovine leukemia virus (BLV) is prevalent worldwide, causing serious problems in the cattle industry. The BLV proviral load (PVL) is a useful index for estimating disease progression and transmission risk. We previously developed a quantitative real-time PCR (qPCR) assay to measure the PVL using the coordination of common motif (CoCoMo) degenerate primers. Here, we constructed a novel duplex BLV-CoCoMo qPCR assay that can amplify two genes simultaneously using a FAM-labeled MGB probe for the BLV LTR gene and a VIC-labeled MGB probe for the BoLA-DRA gene. This liquid duplex assay maintained its original sensitivity and reproducibility in field samples. Furthermore, we developed a dry duplex assay composed of PCR reagents necessary for the optimized liquid duplex assay. We observed a strong positive correlation between the PVLs measured using the dry and liquid duplex assays. Validation analyses showed that the sensitivity of the dry duplex assay was slightly lower than that of the other methods for the detection of a BLV molecular clone, but it showed similar sensitivity to the singleplex assay and slightly higher sensitivity than the liquid duplex assay for the PVL quantification of 82 field samples. Thus, our liquid and dry duplex assays are useful for measuring the BLV PVL in field samples, similar to the original singleplex assay. [ABSTRACT FROM AUTHOR]

Typically, polymer composites and ceramics are used to create biosensors. Materials with properties that are ideal for biosensors and chemical sensors include AgNO3 (silver nitrate), PVDF (polyvinylidene fluoride), and HA (hydroxyapatite). Polyvinylidene fluoride (PVDF) polymer has been widely used in several applications because of its well-known superior ferroelectric characteristics and biocompatibility. The brittleness and low bending strength of hydroxyapatite limit its applicability. Several HA and polymer composite formulations have been developed to compensate for HA's mechanical weakness. The final product contains a significant amount of HA, making HA/polymer composites highly biocompatible. When the right amount of silver is deposited, the maximum piezoelectric activity is generated, and silver nitrate has antimicrobial properties. The non-toxic solvent DMSO (dimethyl sulfoxide) and the solvent casting method were chosen for the preparation of the film. Surface roughness was chosen to measure the Str and Sdr properties of the thin film. For liquid preparation, the multifractal spectra analysis was chosen for each sample. SEM was used to examine the samples morphologically. EDX and mapping analyses were presented for chemistry distribution in the samples. [ABSTRACT FROM AUTHOR]

The Ornstein–Zernike integral equations for thermodynamically equilibrium liquids are considered, taking into account irreducible diagrams. An algorithm for calculating the first irreducible diagram is formulated for a spatially homogeneous fluid. For liquids bordering a solid surface, an equation is obtained that implicitly considers all irreducible diagrams. [ABSTRACT FROM AUTHOR]

In this work, the successful synthesis of nanostructures based on MXenes by initiating a pulse discharge between titanium wires immersed in carbon tetrachloride is presented. By choosing carbon tetrachloride as the environment, the formation of oxide structures in the synthesized samples could be avoided. The characterization of the obtained structures was carried out using various techniques, including scanning electron microscopy, X-ray phase analysis, EDS, Raman and FTIR spectroscopy. The results confirmed the successful formation of MXenes with a Cl− occupying the interlayer space. The properties of the discharge initiated in CCl4 were also determined. The results suggest the potential of this plasma-liquid method for the synthesis of high-purity MXenes without impurities commonly found in conventionally prepared samples. [ABSTRACT FROM AUTHOR]

The detection of the liquid-to-ice transition is an important challenge for many applications. In this paper, a method for multi-parameter characterization of the liquid-to-ice phase transition is proposed and tested. The method is based on the fundamental properties of bulk acoustic waves (BAWs). BAWs with shear vertical (SV) or shear horizontal (SH) polarization cannot propagate in liquids, only in solids such as ice. BAWs with longitudinal (L) polarization, however, can propagate in both liquids and solids, but with different velocities and attenuations. Velocities and attenuations for L-BAWs and SV-BAWs are measured in ice using parameters such as time delay and wave amplitude at a frequency range of 1–37 MHz. Based on these measurements, relevant parameters for Rayleigh surface acoustic waves and Poisson's modulus for ice are determined. The homogeneity of the ice sample is also detected along its length. A dual sensor has been developed and tested to analyze two-phase transitions in two liquids simultaneously. Distilled water and a 0.9% solution of NaCl in water were used as examples. [ABSTRACT FROM AUTHOR]

The elasto-optic properties of liquids on the basis of the first principles of acousto-optics were theoretically investigated. A relationship for calculating the elasto-optic constant of liquids using only the refractive index was obtained. The refractive index values corresponding to the maximum elasto-optic constant for polar and nonpolar liquids were determined. Calculations for about 100 liquids were performed and compared with known experimental data. This study significantly extends our understanding of the acousto-optic effect and has practical applications for predicting the elasto-optic constant of a liquid and estimating its wavelength dispersion. [ABSTRACT FROM AUTHOR]

A review of the author's data, partly still unpublished, on studying of liquid tellurium and cesium is given. No definite proofs indicating phase transitions in these liquids were found. New developments in studying the liquid–liquid phase transition are briefly described. [ABSTRACT FROM AUTHOR]

Fragile X-associated Tremor/Ataxia Syndrome (FXTAS) is a neurodegenerative disorder associated with the FMR1 premutation. Currently, it is not possible to determine when and if individual premutation carriers will develop FXTAS. Thus, with the aim to identify biomarkers for early diagnosis, development, and progression of FXTAS, along with associated dysregulated pathways, we performed blood proteomic profiling of premutation carriers (PM) who, as part of an ongoing longitudinal study, emerged into two distinct groups: those who developed symptoms of FXTAS (converters, CON) over time (at subsequent visits) and those who did not (non-converters, NCON). We compared these groups to age-matched healthy controls (HC). We assessed CGG repeat allele size by Southern blot and PCR analysis. The proteomic profile was obtained by liquid chromatography mass spectrometry (LC-MS/MS). We identified several significantly differentiated proteins between HC and the PM groups at Visit 1 (V1), Visit 2 (V2), and between the visits. We further reported the dysregulated protein pathways, including sphingolipid and amino acid metabolism. Our findings are in agreement with previous studies showing that pathways involved in mitochondrial bioenergetics, as observed in other neurodegenerative disorders, are significantly altered and appear to contribute to the development of FXTAS. Lastly, we compared the blood proteome of the PM who developed FXTAS over time with the CSF proteome of the FXTAS patients recently reported and found eight significantly differentially expressed proteins in common. To our knowledge, this is the first report of longitudinal proteomic profiling and the identification of unique biomarkers and dysregulated protein pathways in FXTAS.

Non-targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a widely used tool for metabolomics analysis, enabling the detection and annotation of small molecules in complex environmental samples. Data-dependent acquisition (DDA) of product ion spectra is thereby currently one of the most frequently applied data acquisition strategies. The optimization of DDA parameters is central to ensuring high spectral quality, coverage, and number of compound annotations. Here, we evaluated the influence of 10 central DDA settings of the Q Exactive mass spectrometer on natural organic matter samples from ocean, river, and soil environments. After data analysis with classical and feature-based molecular networking using MZmine and GNPS, we compared the total number of network nodes, multivariate clustering, and spectrum quality-related metrics such as annotation and singleton rates, MS/MS placement, and coverage. Our results show that automatic gain control, microscans, mass resolving power, and dynamic exclusion are the most critical parameters, whereas collision energy, TopN, and isolation width had moderate and apex trigger, monoisotopic selection, and isotopic exclusion minor effects. The insights into the data acquisition ergonomics of the Q Exactive platform presented here can guide new users and provide them with initial method parameters, some of which may also be transferable to other sample types and MS platforms.

Roles of different ecological classes of algal exometabolites in regulating microbial community composition are not well understood. Here, we identify exometabolites from the model diatom Phaeodactylum tricornutum and demonstrate their potential to influence bacterial abundances. We profiled exometabolites across a time course of axenic algal growth using liquid chromatography-tandem mass spectrometry. We then investigated growth of 12 bacterial isolates on individual-identified exometabolites. Lastly, we compared responses of a P. tricornutum-adapted enrichment community to additions of two contrasting metabolites: selective growth substrate 4-hydroxybenzoic acid and putative signaling/facilitator molecule lumichrome. We identified 50 P. tricornutum metabolites and found distinct temporal accumulation patterns. Two exometabolites (of 12 tested) supported growth of distinct subsets of bacterial isolates. While algal exudates and algal presence drove similar changes in community composition compared with controls, exogenous 4-hydroxybenzoic acid addition promoted increased abundances of taxa that utilized it in isolation, and also revealed the importance of factors relating to algal presence in regulating community composition. This work demonstrates that secretion of selective bacterial growth substrates represents one mechanism by which algal exometabolites can influence bacterial community composition and illustrates how the algal exometabolome has the potential to modulate bacterial communities as a function of algal growth.

Osteogenesis is modulated by multiple regulatory networks. Recent studies showed that RNA modifications and their reader, writer, and eraser (RWE) proteins are involved in regulating various biological processes. Few studies, however, were conducted to investigate the functions of RNA modifications and their RWE proteins in osteogenesis. By using LC-MS/MS in parallel-reaction monitoring (PRM) mode, we performed a comprehensive quantitative assessment of 154 epitranscriptomic RWE proteins throughout the entire time course of osteogenic differentiation in H9 human embryonic stem cells (ESCs). We found that approximately half of the 127 detected RWE proteins were down-regulated during osteogenic differentiation, and they included mainly proteins involved in RNA methylation and pseudouridylation. Protein-protein interaction (PPI) network analysis unveiled significant associations between the down-regulated epitranscriptomic RWE proteins and osteogenesis-related proteins. Gene set enrichment analysis (GSEA) of publicly available RNA-seq data obtained from osteogenesis imperfecta patients suggested a potential role of METTL1 in osteogenesis through the cytokine network. Together, this is the first targeted profiling of epitranscriptomic RWE proteins during osteogenic differentiation of human ESCs, and our work unveiled potential regulatory roles of these proteins in osteogenesis. LC-MS/MS data were deposited on ProteomeXchange (PXD039249).

Distinct CD4+ T cell epitopes have been associated with spontaneous control of HIV-1 replication, but analysis of antigen-dependent factors that influence epitope selection is lacking. To examine these factors, we used a cell-free antigen processing system that incorporates soluble HLA-DR (DR1), HLA-DM (DM), cathepsins, and full-length protein antigens for epitope identification by LC-MS/MS. HIV-1 Gag, Pol, Env, Vif, Tat, Rev, and Nef were examined using this system. We identified 35 novel epitopes, including glycopeptides. Epitopes from smaller HIV-1 proteins mapped to regions of low protein stability and higher solvent accessibility. HIV-1 antigens associated with limited CD4+ T cell responses were processed efficiently, while some protective epitopes were inefficiently processed. 55% of epitopes obtained from cell-free processing induced memory CD4+ T cell responses in HIV-1+ donors, including eight of 19 novel epitopes tested. Thus, an in vitro processing system utilizing the components of Class II processing reveals factors influencing epitope selection of HIV-1 and represents an approach to understanding epitope selection from non-HIV-1 antigens.

Animal and epidemiologic studies suggest that there may be adverse health effects from exposure to glyphosate, the most highly used pesticide in the world, and its metabolite aminomethylphosphonic acid (AMPA). Meanwhile, consumption of organic foods (presumably grown free of chemical pesticides) has increased in recent years. However, there have been limited biomonitoring studies assessing the levels of human glyphosate and AMPA exposure in the United States. We examined urinary levels of glyphosate and AMPA in the context of organic eating behavior in a cohort of healthy postmenopausal women residing in Southern California and evaluated associations with demographics, dietary intake, and other lifestyle factors. 338 women provided two first-morning urine samples and at least one paired 24-h dietary recall reporting the previous day's dietary intake. Urinary glyphosate and AMPA were measured using LC-MS/MS. Participants reported on demographic and lifestyle factors via questionnaires. Potential associations were examined between these factors and urinary glyphosate and AMPA concentrations. Glyphosate was detected in 89.9% of urine samples and AMPA in 67.2%. 37.9% of study participants reported often or always eating organic food, 30.2% sometimes, and 32.0% seldom or never. Frequency of organic food consumption was associated with several demographic and lifestyle factors. Frequent organic eaters had significantly lower urinary glyphosate and AMPA levels, but not after adjustment for covariates. Grain consumption was significantly associated with higher urinary glyphosate levels, even among women who reported often or always eating organic grains. Soy protein and alcohol consumption as well as high frequency of eating fast food were associated with higher urinary AMPA levels. In conclusion, in the largest study to date examining paired dietary recall data and measurements of first-void urinary glyphosate and AMPA, the vast majority of subjects sampled had detectable levels, and significant dietary sources in the American diet were identified.

Bacterial-fungal interactions (BFIs) can shape the structure of microbial communities, but the small molecules mediating these BFIs are often understudied. We explored various optimization steps for our microbial culture and chemical extraction protocols for bacterial-fungal co-cultures, and liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed that metabolomic profiles are mainly comprised of fungi derived features, indicating that fungi are the key contributors to small molecules in BFIs. LC-inductively coupled plasma MS (LC-ICP-MS) and MS/MS based dereplication using database searching revealed the presence of several known fungal specialized metabolites and structurally related analogues in these extracts, including siderophores such as desferrichrome, desferricoprogen, and palmitoylcoprogen. Among these analogues, a novel putative coprogen analogue possessing a terminal carboxylic acid motif was identified from Scopulariopsis sp. JB370, a common cheese rind fungus, and its structure was elucidated via MS/MS fragmentation. Based on these findings, filamentous fungal species appear to be capable of producing multiple siderophores with potentially different biological roles (i.e. various affinities for different forms of iron). These findings highlight that fungal species are important contributors to microbiomes via their production of abundant specialized metabolites and that elucidating their role in complex communities should continue to be a priority.

Isomeric molecules are important analytes in many biological and chemical arenas, yet their similarity poses challenges for many analytical methods, including mass spectrometry (MS). Tandem-MS provides significantly more information about isomers than intact mass analysis, but highly similar fragmentation patterns are common and include cases where no unique m/z peaks are generated between isomeric pairs. However, even in such situations, differences in peak intensity can exist and potentially contain additional information. Herein, we present a framework for comparing mass spectra that differ only in terms of peak intensity and include calculation of a statistical probability that the spectra derive from different analytes. This framework allows for confident identification of peptide isomers by collision-induced dissociation, higher-energy collisional dissociation, electron-transfer dissociation, and radical-directed dissociation. The method successfully identified many types of isomers including various d/l amino acid substitutions, Leu/Ile, and Asp/IsoAsp. The method can accommodate a wide range of changes in instrumental settings including source voltages, isolation widths, and resolution without influencing the analysis. It is shown that quantification of the composition of isomeric mixtures can be enabled with calibration curves, which were found to be highly linear and reproducible. The analysis can be implemented with data collected by either direct infusion or liquid-chromatography MS. Although this framework is presented in the context of isomer characterization, it should also prove useful in many other contexts where similar mass spectra are generated.

Plant responses to environmental change are mediated via changes in cellular metabolomes. However,

BackgroundAcetaminophen has been evaluated in horses for treatment of musculoskeletal pain but not as an antipyretic.ObjectivesTo determine the pharmacokinetics and efficacy of acetaminophen compared to placebo and flunixin meglumine in adult horses with experimentally induced endotoxemia.AnimalsEight university owned research horses with experimentally induced endotoxemia.MethodsRandomized placebo controlled crossover study. Horses were treated with acetaminophen (30 mg/kg PO; APAP), flunixin meglumine (1.1 mg/kg, PO; FLU), and placebo (PO; PLAC) 2 hours after administration of LPS. Plasma APAP was analyzed via LC-MS/MS. Serial CBC, lactate, serum amyloid A, heart rate and rectal temperature were evaluated. Serum IL-1β, IL-6, IL-8, IL-10, and TNF-α were evaluated by an equine-specific multiplex assay.ResultsMean maximum plasma APAP concentration was 13.97 ± 2.74 μg/mL within 0.6 ± 0.3 hour after administration. At 4 and 6 hours after treatment, both APAP (P =

Regular, moderate exercise modifies the gut microbiome and contributes to human metabolic and immune health. The microbiome may exert influence on host physiology through the microbial production and modification of metabolites (xenometabolites); however, this has not been extensively explored. We hypothesized that 6 weeks of supervised, aerobic exercise 3×/week (60%-75% heart rate reserve [HRR], 30-60 min) in previously sedentary, lean (n = 14) and obese (n = 10) adults would modify both the fecal and serum xenometabolome. Serum and fecal samples were collected pre- and post-6 week intervention and analyzed by liquid chromatography/tandem mass spectrometry (LC-MS/MS). Linear mixed models (LMMs) identified multiple fecal and serum xenometabolites responsive to exercise training. Further cluster and pathway analysis revealed that the most prominent xenometabolic shifts occurred within aromatic amino acid (ArAA) metabolic pathways. Fecal and serum ArAA derivatives correlated with body composition (lean mass), markers of insulin sensitivity (insulin, HOMA-IR) and cardiorespiratory fitness ( V̇O2max$$ \dot{\mathrm{V}}{\mathrm{O}}_{2\max } $$ ), both at baseline and in response to exercise training. Two serum aromatic microbial-derived amino acid metabolites that were upregulated following the exercise intervention, indole-3-lactic acid (ILA: fold change: 1.2, FDR p

Cell-to-cell transmission and subsequent amplification of pathological proteins promote neurodegenerative disease progression. Most research on this has focused on pathological protein seeds, but how their normal counterparts, which are converted to pathological forms during transmission, regulate transmission is less understood. Here we show in cultured cells that phosphorylation of soluble, nonpathological α-synuclein (α-Syn) at previously identified sites dramatically affects the amplification of pathological α-Syn, which underlies Parkinson's disease and other α-synucleinopathies, in a conformation- and phosphorylation site-specific manner. We performed LC-MS/MS analyses on soluble α-Syn purified from Parkinson's disease and other α-synucleinopathies, identifying many new α-Syn post-translational modifications (PTMs). In addition to phosphorylation, acetylation of soluble α-Syn also modified pathological α-Syn transmission in a site- and conformation-specific manner. Moreover, phosphorylation of soluble α-Syn could modulate the seeding properties of pathological α-Syn. Our study represents the first systematic analysis how of soluble α-Syn PTMs affect the spreading and amplification of pathological α-Syn, which may affect disease progression.

Carbohydrates are the most abundant biomolecules in nature, and specifically, polysaccharides are present in almost all plants and fungi. Due to their compositional diversity, polysaccharide analysis remains challenging. Compared to other biomolecules, high-throughput analysis for carbohydrates has yet to be developed. To address this gap in analytical science, we have developed a multiplexed, high-throughput, and quantitative approach for polysaccharide analysis in foods. Specifically, polysaccharides were depolymerized using a nonenzymatic chemical digestion process followed by oligosaccharide fingerprinting using high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS). Both label-free relative quantitation and absolute quantitation were done based on the abundances of oligosaccharides produced. Method validation included evaluating recovery for a range of polysaccharide standards and a breakfast cereal standard reference material. Nine polysaccharides (starch, cellulose, β-glucan, mannan, galactan, arabinan, xylan, xyloglucan, chitin) were successfully quantitated with sufficient accuracy (5-25% bias) and high reproducibility (2-15% CV). Additionally, the method was used to identify and quantitate polysaccharides from a diverse sample set of food samples. Absolute concentrations of nine polysaccharides from apples and onions were obtained using an external calibration curve, where varietal differences were observed in some of the samples. The methodology developed in this study will provide complementary polysaccharide-level information to deepen our understanding of the interactions of dietary polysaccharides, gut microbial community, and human health.