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Topic:Physiology

The physiology of horses encompasses the study of the biological functions and processes that occur within the equine body. This includes the examination of various systems such as the cardiovascular, respiratory, musculoskeletal, digestive, and nervous systems. Understanding equine physiology is essential for comprehending how horses adapt to different environmental conditions, perform physical activities, and respond to health challenges. Research in this field often focuses on the mechanisms of energy metabolism, thermoregulation, and muscle function during exercise, as well as the physiological responses to stress and disease. This page compiles peer-reviewed research studies and scholarly articles that explore the diverse aspects of equine physiology, providing insights into the biological processes that support the health and performance of horses.
Effects of injection of botulinum toxin type B into the external anal sphincter on anal pressure of horses.
American journal of veterinary research    January 15, 2004   Volume 65, Issue 1 26-30 doi: 10.2460/ajvr.2004.65.26
Adam-Castrillo D, White NA, Donaldson LL, Furr MO.To determine effects on anal pressure of horses after local injection of the external anal sphincter with Clostridium botulinum toxin type B. Methods: 11 healthy adult horses. Methods: Peak and resting anal sphincter pressures were measured with a custom-made rectal probe that was connected to a pressure transducer. Pressures were measured before treatment and after injection with botulinum toxin type B (BTB) or saline (0.9% NaCl) solution. Dose titration with 500, 1,000, 1,500, and 2,500 U of BTB was completed. Physical changes, behavior, and anal pressure were recorded for each horse. Result...
Evaluation of plasma carboxy-terminal cross-linking telopeptide of type I collagen concentration in horses.
American journal of veterinary research    January 15, 2004   Volume 65, Issue 1 104-109 doi: 10.2460/ajvr.2004.65.104
Carstanjen B, Hoyle NR, Gabriel A, Hars O, Sandersen C, Amory H, Remy B.To evaluate a human assay for quantification of carboxy-terminal cross-linking telopeptide of type I collagen (CTX-I), assess the influence of age on plasma CTX-I concentration, investigate the relationship between plasma CTX-I and serum osteocalcin concentrations, and determine whether concentrations of plasma CTX-I or serum osteocalcin fluctuate in circadian manner in horses. HORSES: 75 clinically normal horses. Methods: Cross-reactivity between equine serum CTX-I and CTX-I antibodies in an automated electrochemiluminescent sandwich antibody assay (ECLIA) was evaluated via a specificity test...
Human chorionic gonadotropin-dependent regulation of 17beta-hydroxysteroid dehydrogenase type 4 in preovulatory follicles and its potential role in follicular luteinization.
Endocrinology    January 15, 2004   Volume 145, Issue 4 1906-1915 doi: 10.1210/en.2003-1715
Brown KA, Boerboom D, Bouchard N, Doré M, Lussier JG, Sirois J.17Beta-hydroxysteroid dehydrogenase type 4 (17betaHSD4) has a unique multidomain structure, with one domain involved in 17beta-estradiol inactivation. The objective of the study was to investigate the regulation of 17betaHSD4 during human chorionic gonadotropin (hCG)-induced ovulation/luteinization. The equine 17betaHSD4 cDNA was cloned and was shown to encode a 735-amino acid protein that is highly conserved (81-87% identity) compared with other mammalian orthologs. RT-PCR/Southern blot analyses were performed to study the regulation of 17betaHSD4 transcripts in equine preovulatory follicles ...
Moments and power generated by the horse (Equus caballus) hind limb during jumping.
The Journal of experimental biology    January 14, 2004   Volume 207, Issue Pt 4 667-674 doi: 10.1242/jeb.00808
Dutto DJ, Hoyt DF, Clayton HM, Cogger EA, Wickler SJ.The ability to jump over an obstacle depends upon the generation of work across the joints of the propelling limb(s). The total work generated by one hind limb of a horse and the contribution to the total work by four joints of the hind limb were determined for a jump. It was hypothesized that the hip and ankle joints would have extensor moments performing positive work, while the knee would have a flexor moment and perform negative work during the jump. Ground reaction forces and sagittal plane kinematics were simultaneously recorded during each jumping trial. Joint moment, power and work wer...
Interaction of fibronectin type II proteins with membranes: the stallion seminal plasma protein SP-1/2.
Biochemistry    January 14, 2004   Volume 43, Issue 2 464-472 doi: 10.1021/bi035647l
Greube A, Müller K, Töpfer-Petersen E, Herrmann A, Müller P.Seminal plasma of mammalians contains, among others, proteins that are characterized by the fibronectin (Fn) type II module. Our knowledge about the structure and the physiological function of seminal Fn type II proteins mainly originates from studies on PDC-109, the bovine representative of this protein family. The present work focuses on the equine protein SP-1/2 (also named HSP-1/2) with particular emphasis on its interaction with lipid membranes by employing the intrinsic protein fluorescence and a number of spin-labeled and fluorescent lipid analogues. The results indicate that the intera...
Critical role of insulin-like growth factor system in follicle selection and dominance in mares.
Biology of reproduction    January 14, 2004   Volume 70, Issue 5 1374-1379 doi: 10.1095/biolreprod.103.026195
Ginther OJ, Gastal EL, Gastal MO, Beg MA.The role of the insulin-like growth factor (IGF) system in the deviation in growth rates among follicles (follicle selection) was studied in mares using an IGF binding protein (BP) to reduce the follicular-fluid concentrations of IGFs. The future dominant follicle (F1) was treated by intrafollicular injection at the expected beginning of deviation (F1 > or = 20 mm; Day 0). The experimental groups were control (no injection, n = 8), vehicle (injection of vehicle; n = 6), and BP (injection of 250 microg of recombinant human IGFBP-3; n = 6). A sample of follicular fluid was taken from F1 on Da...
Scintigraphy of the sacroiliac joint region in asymptomatic riding horses: scintigraphic appearance and evaluation of method. Erichsen C, Eksell P, Widström C, Berger M, Holm KR, Johnston C.The sacroiliac region in 33 clinically normal riding horses was examined with scintigraphy to determine the normal scintigraphic appearance, to evaluate a semiquantitative method used to calculate a sacroiliac joint (SI joint) ratio, and to identify factors that affect the scintigraphic appearance of the pelvis. The scintigraphic examination included dorsal views of each SI joint region and one lateral view of the urinary bladder. Two predefined areas (the SI joint and the area between the tuber sacrale and the SI joint) were evaluated subjectively by comparing the intensity in each area to th...
Effect of trabecular orientation on mechanical resistance and ultrasound propagation in specimens of equine vertebrae.
Ultrasound in medicine & biology    December 31, 2003   Volume 29, Issue 12 1777-1785 doi: 10.1016/j.ultrasmedbio.2003.08.003
Cavani F, Fini M, de Terlizzi F, Cadossi M, Ciminelli L, Ortolani S, Cherubini R, De Aloysio D, Giavaresi G, Cadossi R, Canè V.Osteoporosis involves alterations, not only in density, but also in the architectural organisation of the bone; in particular, trabecular orientation, following the skeletal load directions, lends a high degree of stiffness to the whole bone. We investigated the relationship between trabecular orientation, density, stiffness and ultrasound (US) propagation in two orthogonal directions (par. = parallel to, and ort. = orthogonal to the main orientation of the trabeculae) in cylindrical equine bone specimens (thoracic vertebrae) where a preferential orientation is present. A total of 15 cylinders...
On the ultrasonic properties of tendon.
Ultrasound in medicine & biology    December 31, 2003   Volume 29, Issue 12 1787-1797 doi: 10.1016/s0301-5629(03)01069-x
Garcia T, Hornof WJ, Insana MF.The strong dependence of tendon echogenicity on insonation angle is explored by analyzing echo spectra. Combining echo spectra with high-resolution images from several modalities reveals that fluid spaces surrounding fascicles and bundles are likely sources of ultrasonic scatter. Mathematical models of tendon structure are proposed to explain how the anisotropic microstructure of tendon gives rise to angle-dependent echogenicity. Echo spectra from spontaneously damaged equine tendon samples were compared with normal equine tendon and found to exhibit a dramatic decrease in anisotropic properti...
Blastocyst formation rates in vivo and in vitro of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection.
Biology of reproduction    December 26, 2003   Volume 70, Issue 5 1231-1238 doi: 10.1095/biolreprod.103.023903
Choi YH, Roasa LM, Love CC, Varner DD, Brinsko SP, Hinrichs K.This study was conducted to evaluate in vivo and in vitro development of in vitro-matured equine oocytes fertilized by intracytoplasmic sperm injection. Oocytes were collected from slaughterhouse-derived ovaries, matured in vitro, and injected with frozen-thawed stallion sperm. In vivo development was assessed after transfer of injected oocytes to the oviducts of recipient mares. Mares were killed 7.5-8.5 days after transfer and the uterus and oviducts flushed for embryo recovery. Of 132 injected oocytes transferred, 69 (52%) were recovered; of these, 25 (36%) were blastocysts with a blastocoe...
Spatial and temporal expression of types I and II receptors for transforming growth factor beta in normal equine skin and dermal wounds.
Veterinary surgery : VS    December 23, 2003   Volume 33, Issue 1 70-76 doi: 10.1111/j.1532-950x.2004.04008.x
De Martin I, Theoret CL.To describe immunolocalization of TGF-beta receptors (RI and RII) in normal equine skin and in thoracic or limb wounds, healing normally or with exuberant granulation tissue (EGT). Methods: Group A: six wounds on one metacarpus and one midthoracic area. Group B: six wounds on both metacarpi, one of which was bandaged to stimulate EGT. Immunohistochemistry was used to detect RI and RII expression in wound margins. Methods: Eight horses, randomly assigned to one of two study groups. Methods: Neutralizing polyclonal anti-rabbit RI and RII antibodies were used to detect spatial expression of RI an...
Effects of extracorporeal shock wave therapy on bone.
Veterinary surgery : VS    December 23, 2003   Volume 33, Issue 1 40-48 doi: 10.1111/j.1532-950x.2004.04013.x
McClure SR, Van Sickle D, White MR.Extracorporeal shock waves have been used for 30 years to fragment uroliths for nonsurgical treatment for urolithiasis in humans. Applied to bone, shock waves delivered at the appropriate energy and pulse number, can stimulate osteogenesis. In Europe, shock waves are routinely used to treat nonunions in humans despite poor understanding of the mechanism of action. Shock wave therapy has also been used clinically in horses. Preliminary experimental studies indicate that shock wave therapy does not damage soft tissue in the distal aspect of the equine limb and can stimulate osteogenesis througho...
Effect of focused and radial extracorporeal shock wave therapy on equine bone microdamage.
Veterinary surgery : VS    December 23, 2003   Volume 33, Issue 1 49-55 doi: 10.1111/j.1532-950x.2004.040005.x
Da Costa Gómez TM, Radtke CL, Kalscheur VL, Swain CA, Scollay MC, Edwards RB, Santschi EM, Markel MD, Muir P.To determine whether bone microcracks are altered after application of focused and radial extracorporeal shock wave therapy (ESWT) to the equine distal limb. Methods: An ex vivo experimental model. Methods: A contralateral limb specimen was obtained from 11 Thoroughbred racehorses with a unilateral catastrophic injury. Distal limb specimens were also obtained from 5 non-racing horses. Methods: Three separate skin-covered bone segments were obtained from the mid-diaphysis of the metacarpus (MC3) or metatarsus (MT3). Focused (9,000 shockwaves, 0.15 mJ/mm2, 4 Hz) and radial (9,000 shockwaves, 0.1...
Scanning electron microscopic examination of third metacarpal/third metatarsal bone failure surfaces in thoroughbred racehorses with condylar fracture.
Veterinary surgery : VS    December 23, 2003   Volume 33, Issue 1 2-10 doi: 10.1111/j.1532-950x.2004.04007.x
Stepnik MW, Radtke CL, Scollay MC, Oshel PE, Albrecht RM, Santschi EM, Markel MD, Muir P.To examine the fracture failure surfaces from Thoroughbred horses that had sustained a catastrophic condylar fracture. Methods: Bone specimens from the failure surface were obtained from 12 Thoroughbred racehorses with catastrophic injury and 2 non-racing horses with accidental long bone fracture. Methods: Bone specimens from the failure surface of each fracture were incubated with gold microspheres to label microcracks before examination at x50 to x60,000 using scanning electron microscopy. Microcracking at the failure surface was assessed using a visual analog scale. Results: Branching array...
In vitro inhibition of blood cholinesterase activities from horse, cow, and rat by tetrachlorvinphos.
International journal of toxicology    December 19, 2003   Volume 22, Issue 6 429-433 doi: 10.1177/109158180302200604
Karanth S, Pope C.The organophosphorus insecticide tetrachlorvinphos (TCVP) is commonly used as a feed-through larvicide in many livestock species, including cattle and horses. Cholinesterase (ChE) activity in blood (generally plasma or whole blood) is often employed to assess organophosphorus insecticide intoxication in animals as well as humans. In many species, including horse and man, plasma contains predominantly butyrylcholinesterase whereas red blood cells in all species express exclusively acetylcholinesterase. To evalulate the comparative interaction of TCVP with blood ChEs in different species, we com...
Therapeutic clenbuterol treatment does not alter Ca2+ sensitivity of permeabilized fast muscle fibres from exercise trained or untrained horses.
Journal of muscle research and cell motility    December 18, 2003   Volume 24, Issue 7 471-476 doi: 10.1023/a:1027377731137
Plant DR, Kearns CF, McKeever KH, Lynch GS.Clenbuterol is a beta2-adrenoceptor agonist primarily used for treating bronchospasm and alleviating the symptoms of chronic obstructive pulmonary disease (COPD) in the horse. In other species (rats, mice, sheep, and cattle), chronic high doses of clenbuterol (typically in the milligram per kilogram body weight range) has been shown to cause a muscle directed protein anabolic response. Clenbuterol can also modify muscle fibre composition and therefore potentially affect muscle function. This has implications for the performance of exercising horses being treated with therapeutic doses of clenb...
Validation of a method for collection and assay of pentane in the exhaled breath of the horse.
Research in veterinary science    December 16, 2003   Volume 76, Issue 2 109-112 doi: 10.1016/j.rvsc.2003.08.007
Wyse CA, Love S, Christley RM, Yam PS, Cooper JM, Cumming DR, Preston T.Oxidative stress refers to an imbalance between the production of oxidising free radicals and the antioxidant defenses of the cell, and is associated with many pathogenic processes. Oxidative damage to cellular lipids results in the evolution of pentane and ethane gas, and detection of these hydrocarbons in the exhaled breath can be used to monitor in vivo oxidative stress. The aim of this study was to validate a gas chromatography (GC) method for measurement of breath pentane in the horse. The GC-system developed showed good specificity for discrimination of pentane from other breath hydrocar...
Effects of exercise and glucose administration on content of insulin-sensitive glucose transporter in equine skeletal muscle.
American journal of veterinary research    December 16, 2003   Volume 64, Issue 12 1500-1506 doi: 10.2460/ajvr.2003.64.1500
Lacombe VA, Hinchcliff KW, Devor ST.To characterize insulin-sensitive glucose-transporter (GLUT-4) protein in equine tissues and determine effects of exercise and glucose administration on content of GLUT-4 protein in equine skeletal muscle. Methods: Tissue samples from 9 horses. Methods: Western blot analyses were performed on crude membrane preparations of equine tissues to characterize GLUT-4. In a crossover, randomized study, horses were strenuously exercised for 3 consecutive days and then administered 13.5% glucose or isotonic saline (0.9% NaCl; control) solution, i.v., at similar infusion rates for 12.1 hours. Samples wer...
Changes in heart rate variability in horses during immersion in warm springwater.
American journal of veterinary research    December 16, 2003   Volume 64, Issue 12 1482-1485 doi: 10.2460/ajvr.2003.64.1482
Kato T, Ohmura H, Hiraga A, Wada S, Kuwahara M, Tsubone H.To determine the effects of immersion in warm springwater (38 degrees to 40 degrees C) on autonomic nervous activity in horses. Methods: 10 male Thoroughbreds. Methods: Electrocardiograms were recorded from horses for 15 minutes during a warm springwater bath after being recorded for 15 minutes during stall rest. Variations in heart rate (HR) were evaluated from the power spectrum in terms of low frequency (LF, 0.01 to 0.07 Hz) power and high frequency (HF, 0.07 to 0.6 Hz) power as indices of autonomic nervous activity. Results: Mean (+/- SE) HR during stall rest and immersion in warm springwa...
Evaluation of iohexol clearance used to estimate glomerular filtration rate in clinically normal foals.
American journal of veterinary research    December 16, 2003   Volume 64, Issue 12 1486-1490 doi: 10.2460/ajvr.2003.64.1486
Gonda KC, Wilcke JR, Crisman MV, Ward DL, Robertson JL, Finco DR, Braselton WE.To determine whether pharmacokinetic analysis of data derived from a single i.v. dose of iohexol could be used to predict creatinine clearance and evaluate simplified methods for predicting serum clearance of iohexol with data derived from 2 or 3 blood samples in clinically normal foals. Methods: 10 healthy foals. Methods: Serum disposition of iohexol and exogenous creatinine clearance was determined simultaneously in each foal (5 males and 5 females). A 3-compartment model of iohexol serum disposition was selected via standard methods. Iohexol clearance calculated from the model was compared ...
NaHCO(3) does not affect arterial O(2) tension but attenuates desaturation of hemoglobin in maximally exercising Thoroughbreds.
Journal of applied physiology (Bethesda, Md. : 1985)    December 12, 2003   Volume 96, Issue 4 1349-1356 doi: 10.1152/japplphysiol.01083.2003
Manohar M, Goetz TE, Hassan AS.The objective of the present study was to examine the effects of preexercise NaHCO(3) administration to induce metabolic alkalosis on the arterial oxygenation in racehorses performing maximal exercise. Two sets of experiments, intravenous physiological saline and NaHCO(3) (250 mg/kg i.v.), were carried out on 13 healthy, sound Thoroughbred horses in random order, 7 days apart. Blood-gas variables were examined at rest and during incremental exercise, leading to 120 s of galloping at 14 m/s on a 3.5% uphill grade, which elicited maximal heart rate and induced pulmonary hemorrhage in all horses ...
Relationship between seminal plasma lactoferrin and gonadal function in horses.
The Journal of veterinary medical science    December 11, 2003   Volume 65, Issue 11 1273-1274 doi: 10.1292/jvms.65.1273
Kikuchi M, Takao Y, Tokuda N, Ohnami Y, Orino K, Watanabe K.Total 78 semen samples were obtained from 27 Thoroughbred stallions (aged 6 to 27 years), and were subjected to quantification of lactoferrin (Lf) in seminal plasma and examination of the seminal properties. The seminal plasma Lf concentration varied from 21 to 689 microg/ml, with a mean value of 244 +/- 151 microg/ml (S.D.). The seminal plasma Lf concentration and total seminal plasma Lf positively correlated with the sperm concentration (r=0.5938, P<0.001) and with the total sperm number (r=0.6959, P<0.001), respectively. There was no correlation between seminal plasma Lf and sperm mot...
Interactions of substrate availability, exercise performance, and nutrition with muscle glycogen metabolism in horses.
Journal of the American Veterinary Medical Association    December 11, 2003   Volume 223, Issue 11 1576-1585 doi: 10.2460/javma.2003.223.1576
Lacombe VA, Hinchcliff KW, Taylor LE.No abstract available
Parathyroid hormone (PTH) secretion, PTH mRNA and calcium-sensing receptor mRNA expression in equine parathyroid cells, and effects of interleukin (IL)-1, IL-6, and tumor necrosis factor-alpha on equine parathyroid cell function.
Journal of molecular endocrinology    December 11, 2003   Volume 31, Issue 3 609-620 doi: 10.1677/jme.0.0310609
Toribio RE, Kohn CW, Capen CC, Rosol TJ.Parathyroid hormone (PTH) is secreted by the chief cells of the parathyroid gland in response to changes in ionized calcium (Ca(2+)) concentrations. In this study, we measured PTH secretion, and PTH mRNA and calcium-sensing receptor (CaR) mRNA expression by equine parathyroid chief cells in vitro. We also evaluated the effects of interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF)-alpha on PTH secretion, and PTH and CaR mRNA expression. The relationship between PTH and Ca(2+) was inversely related. PTH secretion decreased from 100% (day 0) to 13% (day 30). PTH mRNA expression decline...
Immunocytochemical localization of cytochrome P450 aromatase in the testis of prepubertal, pubertal, and postpubertal horses.
Theriogenology    December 10, 2003   Volume 61, Issue 2-3 293-299 doi: 10.1016/s0093-691x(03)00237-1
Hess MF, Roser JF.The large amount of testicular estrogens produced by the stallion is unique compared to the amounts found in other domestic species. Although the cellular locale of the cytochrome P450 aromatase (P450arom) enzyme that converts C19 androgens to C18 estrogens has been identified in the Leydig cell of adult equine testis, the location in the immature equine testis is not known. The goal of this work was to localize the enzyme in colts and stallions during sexual development. Testes were obtained from prepubertal (n=7), pubertal (n=6), and postpubertal (n=8) colts and stallions during both the bre...
The effect of acetylsalicylic acid and captopril on uterine and ovarian blood flow during the estrous cycle in mares.
Theriogenology    December 10, 2003   Volume 61, Issue 2-3 301-309 doi: 10.1016/s0093-691x(03)00214-0
Bollwein H, Weber F, Steffen S, Stolla R.In previous studies, transrectal color Doppler sonography was used to demonstrate an increase in genital blood flow resistance in subfertile mares. The objectives of the present study were to determine the effects of an anticoagulant (acetylsalicylic acid) and a vasodilator (captopril) on uterine and ovarian perfusion and plasma progesterone concentrations in cycling mares. From Day 1 to 11 of an estrous cycle (Day 0=day of ovulation following prostaglandin-induced luteolysis), five Trotter mares were given 2500 mg lactose, 2500 mg ASA, or 50 mg captopril twice daily in their feed (one compoun...
Dose-response study of intrafollicular injection of insulin-like growth factor-I on follicular fluid factors and follicle dominance in mares.
Biology of reproduction    December 10, 2003   Volume 70, Issue 4 1063-1069 doi: 10.1095/biolreprod.103.024844
Ginther OJ, Gastal EL, Gastal MO, Checura CM, Beg MA.The effect of insulin-like growth factor-I (IGF-I) on the concentrations of follicular fluid factors during follicle deviation and the development of dominance was studied in mares in two experiments. Transvaginal ultrasound guidance was used for intrafollicular injection and subsequent sequential sampling of follicular fluid. Treatment involved a single injection of IGF-I into the second-largest follicle (F2) at the expected beginning of deviation (Hour 0) based on diameter (> or =20 mm) of the largest follicle (F1). Mares in IGF-I groups were given a dose of 500 microg (experiment 1) or 2...
Factors affecting plasma progesterone concentration and the retrospective determination of time of ovulation in cyclic mares.
Theriogenology    December 10, 2003   Volume 61, Issue 2-3 203-214 doi: 10.1016/s0093-691x(03)00211-5
Nagy P, Huszenicza G, Reiczigel J, Juhász J, Kulcsár M, Abaváry K, Guillaume D.Factors influencing plasma progesterone concentration were investigated in seven mares. Two-phase logistic curves were fitted (r=0.98) to plasma progesterone concentrations of blood samples collected once daily. In addition to the effect of time (P<0.001), there were differences (P<0.01) among mares in the peak height of the progesterone plateau and in the (area under the curve) AUC. Plasma progesterone concentrations were higher (P<0.001) after a multiple versus single ovulation. There was an effect of season (P<0.001), but no significant effect of luteal morphology. The retrospec...
[Enlarged ovary in a mare: review of the literature and a case report].
Tijdschrift voor diergeneeskunde    December 6, 2003   Volume 128, Issue 22 692-696 
Westermann CM, Parlevliet JM, Meertens NM, Sloet van Oldruitenborgh-Oosterbaan MM.Mares regularly have an enlarged ovary. The main causes are haematomas, anovulatory follicles, abscesses, and neoplasia. The granulosa-theca-cell tumour is by far the most common neoplasia of the ovary (about 97%) and accounts for 2.5% of all equine tumours. In this article the differential diagnosis of an enlarged ovary and the background of granulosa-theca cell tumours are reviewed. A case is described of a mare with a very large granulosa-theca cell tumour in the left ovary, which was discovered 1 month after delivery of a healthy foal. This case is special not only because the tumour was e...
Liquid chromatography/electrospray ionization tandem mass spectrometric screening and confirmation methods for beta2-agonists in human or equine urine.
Journal of mass spectrometry : JMS    December 4, 2003   Volume 38, Issue 11 1197-1206 doi: 10.1002/jms.542
Thevis M, Opfermann G, Schänzer W.Electrospray ionization (ESI) mass spectra of 19 common beta(2)-agonists were investigated in terms of fragmentation pattern and dissociation behavior of the analytes, proving the origin of fragment ions and indicating mechanisms of charge-driven and charge-remote fragmentation. Based on these data, liquid chromatographic/ESI tandem mass spectrometric (LC/ESI-MS/MS) screening and confirmation methods were developed for doping control purposes. These procedures employ established sample preparation steps including either acidic or enzymatic hydrolysis, alkaline extraction and, in the case of eq...