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Topic:Mass Spectrometry
Mass spectrometry is an analytical technique used to measure the mass-to-charge ratio of ions, providing detailed information about the molecular composition of samples. In equine research, mass spectrometry is employed to analyze a wide range of biological materials, including blood, tissues, and other bodily fluids. This technique is utilized for various applications, such as identifying biomarkers for disease, studying metabolic pathways, and assessing nutritional and environmental influences on horse health. By enabling precise molecular characterization, mass spectrometry aids in the understanding of equine physiology and pathology. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and findings of mass spectrometry in equine science.
Bioanalytical Uncertainty Assessment of Ultra-High-Performance Liquid Chromatography-High Resolution Mass Spectrometry Method for Caffeine and Lidocaine in Equine Antidoping: A Dual Perspective on Bottom-up and Top-Down Approaches. The quality of quantitative results in bioanalysis requires not only a validated analytical method but also a rigorous estimation of measurement uncertainty. This study examines the challenges associated with the implementation of two distinct approaches in equine anti-doping control for the assessment of uncertainty associated with an ultra-high-performance liquid chromatography-high resolution mass spectrometry quantitative method for caffeine and lidocaine in horse urine. The bottom-up approach, based on the ISO Guide to the Expression of Uncertainty in Measurement (ISO GUM), was compared t...
Plasma and Urine Pharmacokinetics of Intravenous Pridinol in Thoroughbreds for Its Medication Control. We examined the pharmacokinetics of intravenous pridinol in six thoroughbred horses. Each horse received a single 20 mg dose of pridinol mesylate via the jugular vein, and plasma and urine samples were collected over 72 h. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to quantify pridinol concentrations in plasma and urine, allowing for the calculation of pharmacokinetic parameters. A three-compartment model best fit the plasma elimination data. Using the Toutain model, irrelevant plasma and urine concentrations were estimated to be 0.00284 and 0.000612 ng/mL, respec...
Mapping the Metabolic Fate of Suzetrigine in Equine and Camel Models: A Step Toward Reliable Antidoping Detection. Effective pain management remains a persistent challenge, with opioids limited by tolerance, dependence, respiratory depression, and misuse. This has created demand for safer, nonaddictive alternatives. Suzetrigine (VX-548, Journavx) is a novel analgesic with promising efficacy, but its potential misuse in competitive racing requires investigation. Limited information is available on its metabolism and detection, emphasizing the need for metabolic characterization to aid antidoping strategies. Methods: In vitro metabolism of suzetrigine was evaluated using equine liver microsomes and homogeniz...
Differences in Plasma Exposure of Cannabidiol and Cannabidiolic Acid Following Oral Administration to Horses. There has been a growing interest in the use of cannabinoids in horses in recent years. Several studies have reported on the pharmacokinetics of cannabidiol (CBD) in horses. However, cannabidiolic acid (CBDA) has received less attention, despite limited evidence suggesting clinically beneficial effects in other species. Horses were administered 3 mg/kg of CBD, 3 mg/kg of CBDA, and a placebo per os in a crossover design, with a one-week washout period between treatments. Plasma and urine samples were collected and analyzed using ultra high-performance liquid chromatography coupled to tandem...
Proteomic profiling of endometrial tissue and plasma from mares with endometrial fibrosis†. Endometrial fibrosis leads to subfertility when periglandular fibrosis impairs endometrial gland function. Identifying distinct protein expressions in mares suffering from this condition is essential for understanding the pathogenesis and developing treatments to delay fibrotic progression. To gain a better understanding of protein dynamics underlying this condition, Data-Independent Acquisition Mass Spectrometry was utilized to comprehensively characterize and compare the equine endometrial and plasma proteome from mares with different severities of endometrial fibrosis. Mares in diestrus wer...
Identification of Candidate Biomarkers Detected in the Urine of Racehorses After Anabolic Agent Administration: Use of Orthogonal Methods for Structural Elucidation. Biomarker identification by mass spectrometry represents a key step in the workflow of nontargeted metabolomic studies. Given the complexity of the data, this step, which must be carried out by a trained specialist, is time-consuming, and the biomarkers discovered are not always identified. While this stage is not an obstacle to the development of new screening and classification tools, it is nonetheless crucial to a better understanding of the results obtained. For this reason, the aim of this study was to perform structural elucidation of candidate biomarkers, which had previously been displ...
Development and validation of general urine screening method for performance enhancing drugs in racehorses utilizing liquid chromatography-high resolution mass spectrometry (LC-HRMS). The complexity of the drug market and the constant updating of drugs have been challenging issues for drug regulatory authorities. In this manuscript, a high-throughput automated assay based on Liquid Chromatography High Resolution Mass Spectrometry (LC-HRMS) suitable for use as an initial testing procedure covering multiple classes of compounds prohibited in horse racing is described. The assay requires a 250-μL urine aliquot, which is subjected to enzymatic hydrolysis followed by Biotage Isolute supported liquid extraction plates using Biotage Extrahera system, evaporation, and reconstituti...
Unveiling the Xianbei cavalry: a multidisciplinary approach to restore and analyse the first horse-cavalry armour set in China. Between the third and sixth centuries AD (Anno Domini), the Xianbei emerged as a dominant nomadic power in the Eastern Eurasian Steppe, distinguished by their exceptional equestrian culture and the pivotal role of cavalry in warfare. Despite their historical significance, detailed knowledge of their cavalry's weaponry and equipment-particularly armour-remains fragmentary. As a critical element of military technology, armour offers valuable insights into ancient combat strategies and cultural practices. However, research on early Chinese cavalry armour has been constrained by limited textual re...
Suzetrigine in Equestrian Sports: Optimized Extraction and LC-HRMS Detection Strategies. Suzetrigine, a recently approved Na1.8 sodium channel blocker, shows strong potential in the treatment of neurological, psychiatric, and pain-related conditions. Its peripheral selectivity enables effective pain management while avoiding central nervous system complications and addiction risks linked to opioid use. Following FDA approval in January 2025, concerns have emerged regarding its possible misuse for performance enhancement in sports, highlighting the need for reliable detection tools in doping control. Methods: An analytical procedure was designed and validated to detect suzetrigine ...
Horse model of spontaneous atrial fibrillation share proteomic changes with humans. Horses and humans are among the few mammals susceptible to spontaneous atrial fibrillation (AF), both suffering from high recurrence rates after treatment. Treatment resistance is often attributed to progressive atrial remodeling, but current treatment options fail to effectively address this aspect. Here, we introduce a novel horse model of spontaneous AF to investigate the biological pathway changes in early stages of the disease. Through data-independent acquisition mass spectrometry on biopsies from the right and left atrium and left ventricular chamber of horses with early-stage persisten...
Administration study of somapacitan, a long-acting growth hormone derivative, in horse for doping control purpose. Somapacitan is the second generation of recombinant human growth hormone (rhGH) medication that retains the pharmacological effects of rhGH but exhibits a longer duration of action due to its reversible albumin-binding in the body. In general, the use of all recombinant growth hormone (rGH) analogues is banned by the human and animal sports regulatory authorities due to their anabolic and lipolytic effects. However, little is known about the elimination kinetics and biological effects of the newly introduced long-acting rhGH, somapacitan, in horses. This paper describes the administration stud...
Accuracy of two Sepsityper MALDI-TOF MS methods for bacterial identification in bloodstream infections in dogs, foals, and calves using Bayesian latent class model. Accurate diagnosis of bloodstream infections is crucial for survival and antimicrobial de-escalation in veterinary medicine. Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry enables faster identification of bacteria in BSIs. This study aimed to compare diagnostic accuracy of two Sepsityper methods (Rapid Sepsityper and Sepsityper Extraction) with conventional culture method for bacterial identification in clinical samples. Mean time-to-positivity and frequency of bacteria in BSIs were also determined. This diagnostic test study used bloodstream infection samples fro...
Quantitative proteomics unveils potential plasma biomarkers and provides insights into the pathophysiological mechanisms underlying equine metabolic syndrome. Equine Metabolic Syndrome (EMS) is a multifactorial endocrine disorder characterized by obesity, insulin dysregulation (ID), and an increase in the risk of laminitis, a painful condition that can lead to euthanasia in severe cases. Diagnosing EMS is challenging and often relies on clinical history including obesity, difficulty in losing weight, and recurring episodes of laminitis. The gold standard for laboratory support of an EMS diagnosis is the identification of ID, with basal insulin being the simplest and most accessible method, especially in a field setting. However, various factors such...
Faecal microbiota and serum metabolome association with equine metabolic syndrome in connemara ponies. Faecal microbiome and serum metabolome have been studied in human medicine to provide a better understanding of metabolic derangements including diabetes; however, equivalent studies in equine medicine are limited. This was a case-control study conducted to identify differences in faecal microbiota composition and concurrent serum metabolite patterns between metabolically normal Connemara ponies and those with Equine Metabolic Syndrome (EMS). Thirty privately owned Connemara ponies (15 EMS and 15 controls) were included in the study. EMS was diagnosed by oral sugar test (OST). Blood samples we...
Exposure of horses to biotoxins, phytoestrogens, and pesticides from different feed materials and supplementary feeds. The occurrence of biotoxins and chemical residues in marketed horse feeds has direct influences on horse health but has not been studied yet. Objective: The study investigated the exposure and health implications of contaminants in various horse feedstuffs available on the European market. Methods: A total of 108 feed samples representing diverse product categories such as hay, processed roughage products, grains, and various supplementary feeds were collected from different European countries and analyzed for contaminants, including mycotoxins, phytoestrogens, pesticides, and veterinary drug ...
Analytical Data Review on an Artificial Intelligence Platform for Doping Control in Horse Racing. In the screening of prohibited substances (PS) in horse biological samples with gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS) for doping control, an enormous number of chromatograms are generated. Reviewing these chromatograms to identify suspicious findings requires an extensive manual effort. Recent advancements in Artificial Intelligence (AI) enable its use to classify images into different categories. This can potentially be utilized to perform first-line analysis of chromatograms, which are usually displayed as images, by classifying them...
Alterations in the Peritoneal Fluid Proteome of Horses with Colic Attributed to Ischemic and Non-Ischemic Intestinal Disease. Peritoneal fluid (PF) is intimately associated with the gastrointestinal tract, and changes in the PF may directly reflect abdominal pathology. We aimed to quantify differences in the PF proteome between intestinal lesion type (ischemic vs. non-ischemic) and location (small vs. large intestine). PF samples were collected at hospital admission from horses presenting for abdominal pain (colic). Cases were clinically categorized by lesion type and location after resolution (10 per group). PF proteins were extracted and quantified by label-free liquid chromatography-tandem mass spectroscopy. Data ...
Effects of BCAA supplementation on plasma and mare’s milk amino acid contents in Yili mares and growth performance of suckling foals. Branched-chain amino acids (BCAAs) play a crucial role in regulating nutritional metabolism in lactating animals. However, limited research has been conducted on BCAAs in equines. This study aimed to investigate the effects of different doses of BCAA supplementation on plasma and milk amino acid profiles in Yili mares, as well as the growth performance of their suckling foals, thereby providing a scientific basis for optimizing feeding management practices. Eighteen pairs of Yili mares and their sucklings were randomly assigned to four groups: a control group (Group D, no BCAA supplementation)...
Development and validation of a quantitative UHPLC-HRMS bioanalytical method for equine anti-doping control. The quantification of banned substances in equine antidoping control, especially in racehorse urine, necessitates robust analytical methods with high detection levels due to the extremely low concentrations of the target substances and the significant impact of minor variations on doping test results. Reliable quantification is important for substances near regulatory thresholds, which, if exceeded, are prohibited. This study presents the development and validation of a bioanalytical UHPLC-HRMS method for quantifying doping substances in equine urine, including diazepam and acepromazine with a...
Confirmation of a novel, stable estrogen metabolite, as 5a,6a-epoxy-estrone sulfate in stallion blood by LC-MS/MS. Mass spectrometry (MS) has become pivotal for accurately delineating intricate molecular structures for steroids present in minute quantities within biological samples. This study utilized liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS) to identify and characterize a 'new' estrogen metabolite, 5α,6α-epoxy-estrone sulfate, in stallion serum from three animals. The estrogen structure was predicted previously using radiolabeled steroids. HRMS/MS, in combination with a seamless sample preparation involving liquid-liquid extraction and chromatographic separation, enabl...
The Suitability of Dried Blood Spot Sampling for Pharmacokinetic Studies in Veterinary Medicine. Dried blood spot (DBS) sampling has emerged as a promising microsampling technique in biomedical and clinical research, offering advantages such as reduced invasiveness, minimal blood volume requirements, and enhanced analyte stability. Although well established in human medicine for neonatal screening and diagnostic applications, its potential in veterinary pharmacology remains underexplored. This study investigated the feasibility of using DBS samples to quantify anesthetic agents-ketamine and medetomidine in cats and lidocaine in horses-during routine surgical procedures at a veterinary tea...
Multiplex PCR-Liquid Chromatography High-Resolution Tandem Mass Spectrometry for Simultaneous Detection of Multiple Transgenes in Equine Plasma. The development of gene therapy techniques introduces a potential risk of gene doping, which threatens the integrity of sport. In response to this challenge, we have developed a novel analytical method that employs a multiplex polymerase chain reaction (PCR) in conjunction with liquid chromatography high-resolution tandem mass spectrometry (LC-HRMS/MS) for the simultaneous identification of multiple transgenes in equine plasma within a single reaction. The method targets three potential doping transgenes: equine growth hormone 1 (eGH1), equine growth hormone-releasing hormone (eGHRH), and equi...
Synovial fluid alpha-2-macroglobulin, gelsolin and lubricin distinguish between osteoarthritic and healthy equine joints. Synovial fluid (SF) is an ideal sentinel fluid for osteoarthritis (OA) diagnosis and prognostication due to its critical homeostatic role, proximity to articular tissues and immune cell composition. Untargeted proteomics enable identification of soluble markers for diagnostic and therapeutic applications while minimising bias. Objective: To use liquid chromatography-tandem mass spectrometry (LC-MS/MS) to define the SF proteome in horses with and without carpal OA. The goal was to identify differentially regulated proteins in mild-moderate carpal joint disease compared with healthy joints. Meth...
Surrogate analyte approach for the determination of endogenous cortisol, cortisone, DHEA, DHEAS in horse hair and sheep wool by LC-HRMS/MS. Two new quantitative methods for the determination of four stress-related hormones (cortisol, cortisone, dehydroepiandrosterone and dehydroepiandrosterone sulfate) in horse hair and sheep wool by liquid chromatography coupled with hybrid high resolution mass spectrometry were developed and validated. Hormones were extracted overnight from ground samples with methanol after a washing step with the same solvent to remove external contamination. The extract was evaporated and dry residue was solubilized in an acid mixture of methanol and water. The methods were validated according to ICH guidelin...
Population pharmacokinetics and clinical evaluation of intravenous acetaminophen and its metabolites in Andalusian horses. To date, no intravenous pharmacokinetics (PK) studies have assessed acetaminophen or its major metabolites (acetaminophen-glucuronide and acetaminophen-sulphate) in horses. The influence of sex on acetaminophen metabolism in horses is unclear, and Monte Carlo simulations have not been applied to explore potential clinical applications. This study aimed to determine the intravenous PK and safety of acetaminophen in Andalusian horses at 10 and 20 mg/kg, evaluate sex as a covariate, and simulate clinical regimens. Twenty mares and twenty stallions received intravenous acetaminophen, and plasma ...
Distribution of Alprazolam Into the Milk of Lactating Mares and Subsequent Absorption by Nursing Foals. Alprazolam is used to facilitate mare-foal bonding in aggressive or anxious postpartum mares. In humans, alprazolam crosses the blood-milk barrier, but the amount transferred into milk is minor and compatible with breastfeeding as the relative infant dose is < 10%. Similar data are not available for horses. The aim of this study was to measure alprazolam in serum and milk of mares (milk: serum ratio) administered alprazolam, and to determine alprazolam serum concentrations in nursing foals to estimate the extent of absorption. This was a prospective observational study involving 7 healthy...
A Laboratory Protocol for Routine Therapeutic Drug Monitoring of Beta-Lactams Antimicrobials in Horses and Dogs. Background: Although antibiotic resistance is a well-known issue in veterinary medicine, studies proposing real-time therapeutic monitoring (TDM) are lacking. The objective of the present study was to develop a simple and rapid protocol for the real-time therapeutic monitoring of antibiotics in horses and dogs. Methods: A reliable TDM protocol should encompass guidelines for the definition of plasma/serum collection time points, sample management by the clinical staff, transportation to the laboratory, and the availability of robust and swift analytical technologies. Ampicillin and sulbactam w...
Morphine and codeine in racing horse feed: is there reason for concern? Opiates such as morphine and codeine are substances often misused to improve the performance of racing horses during competitions and are therefore on the International Federation for Equestrian Sports' list of prohibited substances. However, a positive antidoping test may be due to the consumption of feed (mainly lucerne or oats) contaminated by opium poppy containing the alkaloids morphine and codeine. In order to determine whether a positive antidoping test is the result of an intentional abuse of opiates or consumption of feed contaminated by poppies, we optimised conditions for the extrac...
Authenticity of domestic animal milk: An identification strategy by supercritical fluid chromatography quadruple time-of-flight mass spectrometry. Livestock milk is important for daily cooking and food production, but some lawless traders often adulterate the milk product using low-priced raw. Therefore, it is urgent to develop a simple, sensitive and accurate fingerprinting identification method to provide consumers with early warning of risks. Herein, we first systematically separated and authenticated the triacylglycerols composition of milks from holstein cattles, goats, mongolian horses, bactrian camels, yaks and buffaloes, which were less reported in domestic and international studies, using the SFC-Q-TOF-MS technique. Subsequently...
Comparative analysis of proteomics and transcriptomics reveals novel mechanism underlying the antibacterial activity and immune-enhancing properties of horse milk. Horse milk is a highly valuable organic food that is a promising alternative to cow milk, exhibiting plenty of healthy and immune benefits to human. However, identification of proteins associated human wellness and underlying molecular mechanism in horse milk remain unclear. Unassigned: Label-free mass spectrometry-based protein quantification technology was employed to investigate protein composition of animal milk, including cow, goat, camel and horse milk. Prokaryotic expression and disk diffusion assay were applied to acquire and evaluate antimicrobial activity of candidate proteins. RAW2...
