Analyze Diet

Topic:Metabolism

Equine metabolism encompasses the biochemical processes that occur within horses to maintain life, including the conversion of food into energy, the synthesis of necessary compounds, and the elimination of waste products. These processes are essential for supporting various physiological functions such as growth, reproduction, and physical performance. Key components of equine metabolism include carbohydrate, fat, and protein metabolism, each of which contributes to the overall energy balance and health of the horse. Factors influencing metabolic rate and efficiency in horses include age, breed, diet, exercise, and health status. This page compiles peer-reviewed research studies and scholarly articles that explore the mechanisms, regulation, and implications of metabolic processes in equine physiology.
Elevated dietary selenium rescues mitochondrial capacity impairment induced by decreased vitamin E intake in young exercising horses.
Journal of animal science    August 1, 2022   Volume 100, Issue 8 skac172 doi: 10.1093/jas/skac172
Owen RN, Semanchik PL, Latham CM, Brennan KM, White-Springer SH.Maintenance of mitochondrial health, which is supported in part by dietary antioxidants such as selenium (Se) and vitamin E (vitE), is pertinent to optimizing athletic performance. Deficiencies in Se and vitE negatively impact muscle health but mitochondrial adaptations to various levels of dietary Se and vitE are poorly understood. Young Quarter Horses (mean ± SD: 17.6 ± 0.9 mo) undergoing submaximal exercise training were used to test the hypothesis that a proprietary antioxidant blend containing elevated Se yeast (EconomasE, Alltech, Inc., Nicholasville, KY) would improve mitochondrial ch...
Differential effect of two dietary protein sources on time course response of muscle anabolic signaling pathways in normal and insulin dysregulated horses.
Frontiers in veterinary science    August 1, 2022   Volume 9 896220 doi: 10.3389/fvets.2022.896220
Loos CMM, McLeod KR, Vanzant ES, Stratton SA, Bohannan AD, Coleman RJ, van Doorn DA, Urschel KL.The objective of the study was to characterize the temporal changes of phosphorylation patterns of mTOR signaling proteins in response to two dietary protein sources in insulin dysregulated (ID, n = 8) and non-ID (n = 8) horses. Horses were individually housed and fed timothy grass hay and 2 daily concentrate meals so that protein was the first limiting nutrient and the total diet provided 120% of daily DE requirements for maintenance. On sample days, horses randomly received 0.25 g CP/kg BW of a pelleted alfalfa (AP) or commercial protein supplement (PS). Blood samples were collected before a...
Variations in the fecal microbiota and their functions of Thoroughbred, Mongolian, and Hybrid horses.
Frontiers in veterinary science    July 28, 2022   Volume 9 920080 doi: 10.3389/fvets.2022.920080
Wen X, Luo S, Lv D, Jia C, Zhou X, Zhai Q, Xi L, Yang C.The horse gut is colonized by a rich and complex microbial community that has important roles in horse physiology, metabolism, nutrition, and immune functions. Fewer across-breed variations in horse gut microbial diversity have been illustrated. In this article, the gut microbiota of Thoroughbred, Mongolian, and Hybrid horses [first filial generation (F1) of Mongolian (maternal) and Thoroughbred (paternal)] were studied by second-generation high-throughput sequencing technology. Differences in gut microbiota composition and function between breeds were determined using diversity and functional...
No size-dependent net particle retention in the hindgut of horses.
Journal of animal physiology and animal nutrition    July 27, 2022   Volume 106, Issue 6 1356-1363 doi: 10.1111/jpn.13757
Schwarm A, Clauss M, Ortmann S, Jensen RB.Sieve analyses of hindgut contents of horses as well as observations in horses where plastic markers had been applied to a caecal cannula suggested that there may be a discrimination by particle size in the passage or retention of digesta. Here, we performed a similar experiment with five caecum-cannulated horses (562 ± 31 kg) fed a constant amount (6.81 kg dry matter/day) of grass hay. Passage markers representing the liquid (Co-EDTA) as well as the particulate digesta phase (Yb-undefined; Cr mordanted fibre 1-2 mm; Ce-mordanted fibre 8 mm) were given as a pulse-dose into the can...
Astaxanthin Carotenoid Modulates Oxidative Stress in Adipose-Derived Stromal Cells Isolated from Equine Metabolic Syndrome Affected Horses by Targeting Mitochondrial Biogenesis.
Biomolecules    July 27, 2022   Volume 12, Issue 8 1039 doi: 10.3390/biom12081039
Mularczyk M, Bourebaba N, Marycz K, Bourebaba L.Astaxanthin is gaining recognition as a natural bioactive component. This study aimed to test whether astaxanthin could protect adipose-derived stromal stem cells (ASCs) from apoptosis, mitochondrial dysfunction and oxidative stress. was used to extract astaxanthin, whose biocompatibility was tested after 24, 48 and 72 h of incubation with the cells; no harmful impact was found. ASCs were treated with optimal concentrations of astaxanthin. Several parameters were examined: cell viability, apoptosis, reactive oxygen levels, mitochondrial dynamics and metabolism, superoxide dismutase activity, ...
Effect of Weight Change on Markers of Bone Turnover and Phosphorus Excretion.
Journal of equine veterinary science    July 16, 2022   Volume 118 104080 doi: 10.1016/j.jevs.2022.104080
Fowler AL, Pyles MB, Hayes SH, Crum AD, Harris PA, Krotky A, Lawrence LM.There is little information about how weight change in horses impacts bone turnover and the metabolism of minerals associated with bone. This study evaluated weight change in mature horses as a factor that could alter bone turnover and fecal P output. Fifteen horses (555 ± 8 kg) were assigned to three treatments: weight loss (LO; n = 5), weight maintenance (MA; n = 5), and weight gain (GA; n = 5). Diets contained 75%, 100%, and 145% of maintenance digestible energy requirements for the three treatments, respectively, but contained similar amounts of protein and minerals. At the end of t...
Simultaneous quantification of caffeine and its main metabolites by gas chromatography mass spectrometry in horse urine.
Biomedical chromatography : BMC    July 15, 2022   Volume 36, Issue 10 e5445 doi: 10.1002/bmc.5445
Göktaş EF, Kabil E, Yatanaslan L, Güneş E, Dirikolu L.Caffeine is a naturally occurring alkaloid and it is metabolized to paraxanthine, theophylline and theobromine. Analysis of caffeine and its metabolites is challenging since the metabolites theophylline and paraxanthine generate similar product and precursor ions. In this study, a new method was developed for the simultaneous analysis of caffeine, paraxanthine, theobromine and theophylline in horse urine using gas chromatography-mass spectrometry (GC-MS). Urine samples were treated using solid-phase extraction followed by the elution with dichloromethane-isopropanol (90:10) after the pH was ad...
Investigating the Relationship Between Cardiac Function and Insulin Sensitivity in Horses: A Pilot Study.
Frontiers in veterinary science    July 8, 2022   Volume 9 899951 doi: 10.3389/fvets.2022.899951
Williams NJ, Furr M, Navas de Solis C, Campolo A, Davis M, Lacombe VA.Metabolic syndrome in humans is commonly associated with cardiovascular dysfunction, including atrial fibrillation and left ventricular diastolic dysfunction. Although many differences exist between human and equine metabolic syndrome, both of these conditions share some degree of insulin resistance. The aims of this pilot study were to investigate the relationship between insulin sensitivity and cardiac function. Seven horses (five mares, two geldings, aged 17.2 ± 4.2 years, weight 524 ± 73 kg) underwent insulin-modified frequently sampled intravenous glucose tolerance testing to determine ...
Serum bone metabolism biomarkers in healthy filies and colts from weaning until one year of age.
Research in veterinary science    July 6, 2022   Volume 150 156-163 doi: 10.1016/j.rvsc.2022.06.018
Arfuso F, Assenza A, Tosto F, Giannetto C, Interlandi C, Piccione G, Liotta L.This study aimed to investigate the serum concentration of osteocalcin (OC), parathyroid hormone (PTH), calcitonin (CT), calcium (Ca), phosphorus (P), alkaline phosphatase (ALP) and acid phosphatase (AP) in healthy-weaned-foals from 8 months of age until the first year of life. Moreover, the correlation of investigated parameters and foals' age, as well as the relationship between the serum values of PTH and the other markers of bone remodeling were assessed. From 20 foals (10 fillies, 10 colts), blood samples were monthly collected (from 8 to 12 months of age, T1-T5) and the concentration o...
Equus roundworms (Parascaris univalens) are undergoing rapid divergence while genes involved in metabolic as well as anthelminic resistance are under positive selection.
BMC genomics    July 4, 2022   Volume 23, Issue 1 489 doi: 10.1186/s12864-022-08702-6
Han L, Lan T, Lu Y, Zhou M, Li H, Lu H, Wang Q, Li X, Du S, Guan C, Zhang Y, Sahu SK, Qian P, Zhang S, Zhou H, Guo W, Chai H, Wang S, Liu Q, Liu H....The evolution of parasites is often directly affected by the host's environment. Studies on the evolution of the same parasites in different hosts are of great interest and are highly relevant to our understanding of divergence. Methods: Here we performed whole-genome sequencing of Parascaris univalens from different Equus hosts (horses, zebras and donkeys). Phylogenetic and selection analyses were performed to study the divergence and adaptability of P. univalens. Results: At the genetic level, multiple lines of evidence indicate that P. univalens is mainly separated into two clades (horse-de...
Short communication: Utilisation of n-alkanes to estimate feed intake in horses fed known amounts of a labelled concentrate supplement.
Animal : an international journal of animal bioscience    June 22, 2022   Volume 16, Issue 7 100571 doi: 10.1016/j.animal.2022.100571
Maxfield M, Celaya R, Andrade E, Rodrigues MAM, Santos AS, Ferreira LMM.The assessment of feed intake in stabled horses is a difficult task to accomplish. Faecal markers, namely n-alkanes, have been used successfully for the estimation of this important nutritional parameter. This usually involves the dosing of synthetic n-alkanes via different matrices, a laborious task that may also influence the animal normal foraging behaviour. An experiment was conducted to evaluate a relative simple methodology to quantify feed intake in horses, based on the provision of measured amounts of a concentrate supplement labelled with beeswax and the utilisation of n-alkanes as fa...
Next-Generation Sequencing analysis discloses genes implicated in equine endometrosis that may lead to tumorigenesis.
Theriogenology    June 20, 2022   Volume 189 158-166 doi: 10.1016/j.theriogenology.2022.06.015
Witkowski M, Duliban M, Rak A, Profaska-Szymik M, Gurgul A, Arent ZJ, Galuszka A, Kotula-Balak M.Endometrosis is a periglandular fibrosis associated with dysfunction of affected glandular epithelial cells that is the most common cause of reduced fertility in mares, although it is not fully understood. The etiology of the disease is still partially unknown. This study focuses on understanding the genetic mechanisms potentially underlying endometrosis in mares using the Next Generation Sequencing (NGS) technique. Endometrial samples, used in the study, were obtained in the anestrus phase both from healthy mares and those diagnosed with endometrosis. The NGS data were analyzed for gene invol...
Characterization of growth hormone secretagogue small molecule ibutamoren (MK-0677) and its possible metabolites in thoroughbred horses for doping control.
Rapid communications in mass spectrometry : RCM    June 19, 2022   Volume 36, Issue 18 e9337 doi: 10.1002/rcm.9337
Philip M, Karakka Kal AK, Subhahar MB, Karatt TK, Mathew B, Koshy SA.It is important to remember that performance-enhancing agents such as non-peptide growth hormone secretagogues present a significant risk of abuse. Ibutamoren (MK-0677) is a potent, long-acting, selective non-peptide growth hormone secretagogue that can be taken orally. Methods: This study examines ibutamoren and its metabolites in thoroughbred horses after oral administration. Liquid chromatography/high-resolution mass spectrometry was used to determine the probable structures of the detected metabolites. Results: In this study, 22 metabolites of ibutamoren were identified (17 phase I and 5 p...
Influence of Azadirachta indica and Cnidoscolus angustidens Dietary Extracts on Equine Fecal Greenhouse Gas Emissions.
Journal of equine veterinary science    June 16, 2022   Volume 116 104049 doi: 10.1016/j.jevs.2022.104049
Alvarado TD, Elghandour MMMY, Ekanem NJ, Alcala-Canto Y, Velázquez AE, Pacheco EBF, Purba RAP, Salem AZM.The present study was conducted to investigate the aqueous extracts of Azadirachta indica (AZN), Cnidoscolus angustidens (CNA), and their combination (MIX) at dosages of 0-, 0.6-, 1.2-, and 1.8- mL for their ability to reduce greenhouse gases and fermentation profiles in an in vitro study using horse feces and a nutrient-dense diet (as substrate). The quantity of greenhouse gas and fermentation profiles were determined in in vitro incubation for 48 h. Extracts of AZN, CNA, and MIX reduced total gas production of the incubated and degraded substrates in a dose-dependent and time-dependent manne...
An exercise-inducible metabolite that suppresses feeding and obesity.
Nature    June 15, 2022   Volume 606, Issue 7915 785-790 doi: 10.1038/s41586-022-04828-5
Li VL, He Y, Contrepois K, Liu H, Kim JT, Wiggenhorn AL, Tanzo JT, Tung AS, Lyu X, Zushin PH, Jansen RS, Michael B, Loh KY, Yang AC, Carl CS....Exercise confers protection against obesity, type 2 diabetes and other cardiometabolic diseases1-5. However, the molecular and cellular mechanisms that mediate the metabolic benefits of physical activity remain unclear6. Here we show that exercise stimulates the production of N-lactoyl-phenylalanine (Lac-Phe), a blood-borne signalling metabolite that suppresses feeding and obesity. The biosynthesis of Lac-Phe from lactate and phenylalanine occurs in CNDP2+ cells, including macrophages, monocytes and other immune and epithelial cells localized to diverse organs. In diet-induced obese mice,...
Oxidant and Antioxidant Parameters’ Assessment Together with Homocysteine and Muscle Enzymes in Racehorses: Evaluation of Positive Effects of Exercise.
Antioxidants (Basel, Switzerland)    June 15, 2022   Volume 11, Issue 6 1176 doi: 10.3390/antiox11061176
This study aimed to evaluate the changes in serum oxidant and antioxidant parameters together with the serum values of homocysteine (Hcy) and muscle enzymes including creatine kinase (CK), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) in regularly trained athletic horses during official races of 1200, 1600 and 2000 m. Thirty Thoroughbred horses were divided into three groups of 10 subjects each according to the race distance: Group 1, 1200 m race; Group 2, 1600 m race; Group 3, 2000 m race. Blood samples were collected from horses 1 week prior to the race (1WB), on the day o...
Fueling equine performance: importance of mitochondrial phenotype in equine athletes.
Animal frontiers : the review magazine of animal agriculture    June 14, 2022   Volume 12, Issue 3 6-14 doi: 10.1093/af/vfac023
Latham CM, Guy CP, Wesolowski LT, White-Springer SH.No abstract available
importance of mitochondrial phenotype in equine athletes.
Animal frontiers : the review magazine of animal agriculture    June 14, 2022   Volume 12, Issue 3 5 doi: 10.1093/af/vfac032
No abstract available
Evaluation of peripheral blood polymorphonuclear cell functions after an oral carbohydrate overload in obese and insulin dysregulated horses.
Veterinary immunology and immunopathology    June 13, 2022   Volume 250 110455 doi: 10.1016/j.vetimm.2022.110455
Salinas C, Espinosa G, Morales N, Henríquez C, Morán G, Gajardo G, Burgos RA, Uberti B.Obesity and insulin dysregulation (ID) are increasingly prevalent conditions in equid populations worldwide. Immune impairment is well described in humans with metabolic dysfunction and is reported but still incompletely understood in horses. This study evaluated the effect of acute induced transient hyperglycemia on apoptosis, phagocytosis and oxidative burst activity of peripheral blood polymorphonuclear cells (PMN) of lean and obese adult horses with or without insulin dysregulation. Seventeen adult horses were allocated into three groups based on their body condition score (BCS) and metabo...
Applicability of the auricular temperature for the assessment of body temperature in healthy large and small domestic species, in a normal metabolic state and in controlled environmental conditions.
Journal of thermal biology    June 9, 2022   Volume 108 103281 doi: 10.1016/j.jtherbio.2022.103281
Giannetto C, Di Pietro S, Pennisi M, Acri G, Piccione G, Giudice E.In clinical practice, an important aspect of animal health status evaluation is the body temperature recording obtained using different methods. Within them, the non-contact and non-invasive infrared thermometer can provide an accurate estimation of body temperature improving the quality of care and medical decision. To evaluate the use of the monitoring of auricular temperature to improve the quality of care and medical decision. To evaluate the use of the monitoring of auricular temperature as indices of body temperature, as well as rectal temperature in eleven clinically healthy mixed bred ...
Comparative Analysis of Microbiome Metagenomics in Reintroduced Wild Horses and Resident Asiatic Wild Asses in the Gobi Desert Steppe.
Microorganisms    June 7, 2022   Volume 10, Issue 6 1166 doi: 10.3390/microorganisms10061166
Tang L, Gao Y, Yan L, Jia H, Chu H, Ma X, He L, Wang X, Li K, Hu D, Zhang D.The gut microbiome offers important ecological benefits to the host; however, our understanding of the functional microbiome in relation to wildlife adaptation, especially for translocated endangered species, is lagging. In this study, we adopted a comparative metagenomics approach to test whether the microbiome diverges for translocated and resident species with different adaptive potentials. The composition and function of the microbiome of sympatric Przewalski's horses and Asiatic wild asses in desert steppe were compared for the first time using the metagenomic shotgun sequencing approach....
Diagnostic evaluation of insulin and glucose dynamics in light-breed horses receiving dexamethasone.
The Canadian veterinary journal = La revue veterinaire canadienne    June 4, 2022   Volume 63, Issue 6 617-626 
Timko KJ, Hostnik LD, Watts MR, Chen C, Bercz A, Toribio RE, Belknap JK, Burns TA.Insulin dysregulation is a hallmark of equine metabolic syndrome (EMS) and increases the risk for development of laminitis. Accurate diagnosis of insulin dysregulation is crucial for implementation of preventative strategies in this population. The objective was to assess the effects of dexamethasone administration on insulin and glucose dynamics in light-breed horses and assess the agreement of various diagnostic tests for insulin dysregulation [basal [insulin] (BI), oral sugar test (OST), and combined glucose-insulin test (CGIT)]. Fourteen adult light-breed horses. Prospective, experimental ...
Effects of Bromocriptine on Glucose and Insulin Dynamics in Normal and Insulin Dysregulated Horses.
Frontiers in veterinary science    May 31, 2022   Volume 9 889888 doi: 10.3389/fvets.2022.889888
Loos CMM, Urschel KL, Vanzant ES, Oberhaus EL, Bohannan AD, Klotz JL, McLeod KR.The objectives of the study were to study the effects of the synthetic ergot alkaloid (EA), bromocriptine, on glucose and lipid metabolism in insulin dysregulated (ID, n = 7) and non-ID (n = 8) mares. Horses were individually housed and fed timothy grass hay and two daily concentrate meals so that the total diet provided 120% of daily DE requirements for maintenance. All horses were given intramuscular bromocriptine injections (0.1 mg/kg BW) every 3 days for 14 days. Before and after 14 days of treatment horses underwent a combined glucose-insulin tolerance test (CGIT) to assess insulin sensit...
Pharmacokinetics, adverse effects and effects on thermal nociception following administration of three doses of codeine to horses.
BMC veterinary research    May 25, 2022   Volume 18, Issue 1 196 doi: 10.1186/s12917-022-03299-0
Knych HK, Stucker K, Gretler SR, Kass PH, McKemie DS.In humans, codeine is a commonly prescribed analgesic that produces its therapeutic effect largely through metabolism to morphine. In some species, analgesic effects of morphine have also been attributed to the morphine-6-glucuronide (M6G) metabolite. Although an effective analgesic, administration of morphine to horses produces dose-dependent neuroexcitation at therapeutic doses. Oral administration of codeine at a dose of 0.6 mg/kg has been shown to generate morphine and M6G concentrations comparable to that observed following administration of clinically effective doses of morphine, witho...
A high-starch vs. high-fibre diet: effects on the gut environment of the different intestinal compartments of the horse digestive tract.
BMC veterinary research    May 19, 2022   Volume 18, Issue 1 187 doi: 10.1186/s12917-022-03289-2
Raspa F, Vervuert I, Capucchio MT, Colombino E, Bergero D, Forte C, Greppi M, Cavallarin L, Giribaldi M, Antoniazzi S, Cavallini D, Valvassori E....Horses are often fed high amounts of starch in their diets despite the well-established benefits of a fibre-based diet to promote gut health and animal welfare. The aim of the present study was to compare the effects of two different diets - one based on high amounts of starch (HS) vs. one base on high amounts of fibre (HF) - on specific parameters of the gut environment across different intestinal compartments of the horse digestive tract. To this end differences in the gastrointestinal environment between HS vs. HF fed horses were assessed in terms of dry matter, organic matter and ash conte...
Allogeneic Serum and Macromolecular Crowding Maintain Native Equine Tenocyte Function in Culture.
Cells    May 5, 2022   Volume 11, Issue 9 1562 doi: 10.3390/cells11091562
Rampin A, Skoufos I, Raghunath M, Tzora A, Diakakis N, Prassinos N, Zeugolis DI.The absence of a native extracellular matrix and the use of xenogeneic sera are often associated with rapid tenocyte function losses during in vitro culture. Herein, we assessed the influence of different sera (equine serum and foetal bovine serum) on equine tenocyte morphology, viability, metabolic activity, proliferation and protein synthesis as a function of tissue-specific extracellular matrix deposition (induced via macromolecular crowding), aging (passages 3, 6, 9) and time in culture (days 3, 5, 7). In comparison to cells at passage 3, at day 3, in foetal bovine serum and without macrom...
Comparative Analysis of Gut Microbiota Between Healthy and Diarrheic Horses.
Frontiers in veterinary science    May 2, 2022   Volume 9 882423 doi: 10.3389/fvets.2022.882423
Li Y, Lan Y, Zhang S, Wang X.Increasing evidence reveals the importance of gut microbiota in animals for regulating intestinal homeostasis, metabolism, and host health. The gut microbial community has been reported to be closely related to many diseases, but information regarding diarrheic influence on gut microbiota in horses remains scarce. This study investigated and compared gut microbial changes in horses during diarrhea. The results showed that the alpha diversity of gut microbiota in diarrheic horses decreased observably, accompanied by obvious shifts in taxonomic compositions. The dominant bacterial phyla (, and )...
High throughput deep proteomic analysis of seminal plasma from stallions with contrasting semen quality.
Systems biology in reproductive medicine    April 28, 2022   Volume 68, Issue 4 272-285 doi: 10.1080/19396368.2022.2057257
Talluri TR, Kumaresan A, Paul N, Sinha MK, Ebenezer Samuel King JP, Elango K, Sharma A, Raval K, Legha RA, Pal Y.Seminal plasma proteins and pathways associated with sperm motility have not been elucidated in stallions. Therefore, in the current study, using the high throughput LC/MS-MS approach, we profiled stallion seminal plasma proteins and identified the proteins and pathways associated with sperm motility. Seminal plasma from six stallions producing semen with contrasting sperm motility ( = 3 each high-and low-motile group) was utilized for proteomic analysis. We identified a total of 1687 proteins in stallion seminal plasma, of which 1627 and 1496 proteins were expressed in high- (HM) and low-...
Causative mechanisms and functional correlates of MTT reduction in stallion spermatozoa.
Reproduction (Cambridge, England)    April 22, 2022   Volume 163, Issue 6 341-350 doi: 10.1530/REP-21-0464
Medica AJ, Gibb Z, Sheridan A, Harrison N, Aitken RJ.MTT is a commonly used cell vitality probe, due to its ability to form insoluble formazan deposits at cellular locations of intense oxidoreductase activity. Although this response is considered a reflection of mitochondrial redox activity, extra-mitochondrial sites of MTT reduction have been recognized within the spermatozoa of several mammalian species. Therefore, the aim of this study was to determine the major sites and causative mechanisms of MTT reduction in stallion spermatozoa. Our results show that stallion spermatozoa displayed substantial mitochondrial formazan deposition, as well as...
A quantitative method for the simultaneous detection of SR9009 and SR9011 in equine plasma using liquid chromatography-electrospray ionization-tandem mass spectrometry.
Drug testing and analysis    April 19, 2022   Volume 14, Issue 8 1532-1538 doi: 10.1002/dta.3271
Kwak YB, Yu J, Yoo HH.SR9009 and SR9011 are metabolic modulators pharmacologically targeting REV-ERB receptors as synthetic agonists. A liquid chromatography-tandem mass spectrometry method for the detection of SR9009 and SR9011 in equine plasma was developed and validated. Plasma samples were pretreated by protein precipitation with methanol and were loaded onto an ACQUITY ultra performance liquid chromatography high-strength silica C18 column (2.1 × 150 mm, 1.8 μm) for chromatographic separation. The mobile phase consisted of 5-mM ammonium formate (pH 3.0) in distilled water and 0.1% formic acid in ac...
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