EQUINE VETERINARY JOURNAL Equine vet. J. (2009) 41 (5) 419-422 doi: 10.

2746/042516409X382079

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General Articles
Pharmacokinetics of detomidine administered to horses at rest and after maximal exercise
J. A. E. HUBBELL*, R. A. SAMS, L. M. SCHMALL, J. T. ROBERTSON, K. W. HINCHCLIFF and W. W. MUIR Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio 43210; and Florida Racing Laboratory, College of Veterinary Medicine, University of Florida, Gainesville, Florida 32607, USA. Keywords: horse; exercise; sedation; detomidine; pharmacokinetics

Summary Reason for performing study: Increased doses of detomidine are required to produce sedation in horses after maximal exercise compared to calm or resting horses. Objectives: To determine if the pharmacokinetics of detomidine in Thoroughbred horses are different when the drug is given during recuperation from a brief period of maximal exercise compared to administration at rest. Methods: Six Thoroughbred horses were preconditioned by exercising them on a treadmill. Each horse ran a simulated race at a treadmill speed that caused it to exercise at 120% of its maximal oxygen consumption. One minute after the end of exercise, horses were treated with detomidine. Each horse was treated with the same dose of detomidine on a second occasion a minimum of 14 days later while standing in a stocks. Samples of heparinised blood were obtained at various time points on both occasions. Plasma detomidine concentrations were determined by liquid chromatographymass spectrometry. The plasma concentration vs. time data were analysed by nonlinear regression analysis. Results: Median back-extrapolated time zero plasma concentration was significantly lower and median plasma half-life and median mean residence time were significantly longer when detomidine was administered after exercise compared to administration at rest. Median volume of distribution was significantly higher after exercise but median plasma clearance was not different between the 2 administrations. Conclusions and potential relevance: Detomidine i.v. is more widely distributed when administered to horses immediately after exercise compared to administration at rest resulting in lower peak plasma concentrations and a slower rate of elimination. The dose requirement to produce an equivalent effect may be higher in horses after exercise than in resting horses and less frequent subsequent doses may be required to produce a sustained effect.

Introduction Detomidine is an 2 receptor agonist that produces sedation, analgesia and muscle relaxation when administered i.v., i.m. or sublingually to horses (Wagner et al. 1991; Ramsay et al. 2002). Sedatives are usually administered to horses at rest to facilitate veterinary procedures but on occasion, sedatives must be administered to excited horses or horses that are injured while exercising to facilitate emergency care. Doses of detomidine (1020 g/kg bwt) that are effective when administered i.v. to calm or resting horses are ineffective when administered immediately after maximal exercise but doubling the dose produces good effect (Hubbell et al. 1997a). The causes of the increased dose requirement are unknown, but the level of central nervous system excitement and the changes in plasma volume and increased cardiac output associated with exercise are likely contributors (Parks and Manohar 1983; Snow et al. 1992; McKeever et al. 1993; Pagliari and Peyrin 1995; Hubbell et al. 1997a,b, 1999). Concentrations of neuroexcitatory hormones increase during exercise, potentially increasing the plasma concentration of sedative required to produce the desired effect (Snow et al. 1992; Pagliari and Peyrin 1995). The pharmacokinetics of 2 clinically relevant i.v. doses of detomidine (10 and 20 g/kg bwt) have been reported (Elfenbein et al. 2009). A 2-compartment model was described with volumes of distribution of 1640 and 1233 ml/kg bwt, mean residence times of 45 and 59 min, and distributive half-lives of 6.4 and 12.2 min after the 10 and 20 g/kg bwt doses, respectively. The dose of detomidine appeared to alter its pharmacokinetics since clearance was decreased and the terminal elimination half-life increased after the larger dose. An earlier study describing the elimination of a very large dose of detomidine (80 g/kg bwt) reported a clearance of 7.1 ml/min/kg bwt and a terminal half-life of 71 min, further supporting arguments for dose dependent elimination (Salonen et al. 1989). The purpose of the present study was to determine if the pharmacokinetics of detomidine in horses immediately after severe exercise are the same as in calm or resting horses.

*Author to whom correspondence should be addressed. Present addresses: Dr Robertson: 2099 Amity Rd., Hilliard, Ohio 43026, USA; Dr Hinchcliff: Veterinary Clinical Centre, University of Melbourne, Werribee, Victoria 3030, Australia; Dr Muir: 338 W 7th Ave., Columbus, Ohio 43201, USA.
[Paper received for publication 22.08.08; Accepted 09.10.08]

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Materials and methods Horses Six mature Thoroughbred horses (2 geldings, 4 mares; mean body weight, 474 kg) were studied. Horses were entered into an exercise regimen designed to establish and maintain a high level of fitness. The described protocols and procedures were approved by the Institutional Animal Care and Use Committee. Study design Horses were exercised on a treadmill1 at a 4 incline, 5 days each week for a minimum of 5 weeks in the following manner: trot 5 min at 4 m/s; walk 2 min at 1.9 m/s; trot 5 min at 4 m/s; walk 2 min at 1.9 m/s; and canter 5 min at 8 m/s. The horses were then tested to determine their individual maximal rate of oxygen consumption (VO2max). The treadmill speed that caused the horse to exercise at 120% of VO2max was calculated and used to simulate racing conditions (Hinchcliff et al. 1993). The 5 day/week exercise programme was continued throughout the course of the study. In addition, each horse was instrumented for measurement of cardiopulmonary and metabolic indices and ran a simulated race at a 4 incline. The simulated race consisted of a warm-up period (trot 5 min at 4 m/s, walk 2 min at 1.9 m/s, canter 5 min at 8 m/s, and walk 2 min at 1.9 m/s) followed by a sprint at the treadmill speed that caused each horse to exercise at 120% of its VO2max. Horses sprinted until fatigued or for a maximum of 120 s. Fatigue was defined as the inability of the horse to maintain its position on the treadmill despite vigorous oral encouragement. One minute after the end of exercise each horse received i.v. detomidine (40 g/kg bwt). Detomidine was administered via an 8 French catheter introducer placed in the left jugular vein. Samples (10 ml) of heparinised blood were obtained prior to exercise and at 5, 15, 30, 45, 60 and 90 min after exercise via a 90 cm, polyethylene 240 catheter2 placed in the right jugular vein and positioned so that its tip was in the right atrium. Correct placement of the catheter was confirmed before and after exercise by visualisation of characteristic pressure waveforms. The catheters were placed to facilitate measurement of cardiopulmonary indices as reported previously (Hubbell et al. 1999). On a second occasion, a minimum of 2 weeks after the initial administration, the horses were placed in a stocks. The hair over both jugular veins was clipped and the skin was prepared for aseptic placement of i.v. catheters after lidocaine (2% 1.5 ml/site) was injected subcut. A 14 gauge catheter was placed in each jugular vein3. Detomidine (40 g/kg bwt) was administered i.v. via the catheter in the left jugular catheter. Ten ml samples of heparinised blood were obtained prior to drug administration and at 5, 15, 30, 45, 60 and 90 min after drug administration via the right jugular catheter. Samples were centrifuged within 60 min after collection, the plasma harvested and transferred to capped vials then frozen at -60C until analysis. Sample analysis

concentrations of detomidine in plasma samples were determined by the internal standard method (medetomidine) using ion peak area ratios and linear regression analysis. The accuracy (% recovery) and corresponding precision (% coefficient of variation) for control samples were 87.6 and 6.5% at 0.25 ng/ml, 105.7 and 8.0% at 0.4 ng/ml, 105.6 and 0.3% at 1 ng/ml, and 103 and 7.1% at 5 ng/ml, respectively. Pharmacokinetic analysis The plasma concentration vs. time data were analysed by nonlinear regression analysis using a computer program5. The plasma detomidine vs. time curves were analysed initially to determine the best pharmacokinetic model that described the data. Based on visual inspection of the agreement between the experimental data and model-predicted values as well as the goodness-of-fit criteria, a one-compartment model was selected. The plasma detomidine concentrations were weighted by 1/Cp2. Parameters calculated in the analysis were Cp0, the elimination rate constant, the elimination half-life (t), AUC090 and AUC0. Furthermore, values for the volume of distribution and clearance of detomidine for each exercised and nonexercised trial were calculated. All parameters were calculated separately for each horse and trial and the median values and ranges were determined for each trial. Statistical analysis The plasma detomidine concentrations are graphed as mean and s.d. of the mean. The data were not normally distributed. Pharmacokinetic values are reported as median values and range. Differences between values were calculated using the Wilcoxon Signed Rank Test. Level of significance was set at P<0.05.
TABLE 1: Pharmacokinetics of detomidine in 6 horses at rest and after exercise Variable At rest After exercise

Median time zero concentration (ng/ml) 68 (4683) 30* (2535) Median half-life (min) 26 (2128) 46* (3954) Median volume of distribution (ml/kg bwt) 585 (481877) 1296* (11311624) Median mean residence time (min) 37 (3049) 66* (5678) Median clearance (ml/min/kg bwt) 16 (1520) 20 (1622) Values are expressed as the median and range. *Indicates values that are significantly different (P<0.05).

80

Detomidine (ng/ml)

60

40

20

Plasma detomidine concentrations were determined by liquid chromatography-mass spectrometry4 (LC-MS) under electrospray ionisation conditions in the positive ion MS/MS mode of operation using a previously described method (Elfenbein et al. 2009). The

20

40 60 Time (min)

80

100

Fig 1: Plasma detomidine concentration in 6 horses given 40 g/kg bwt i.v., at rest and after exercise. Means 1 s.d. l = Resting; n = Exercised.

J. A. E. Hubbell et al.

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Results The median back-extrapolated time zero plasma concentration of detomidine was significantly lower after exercise than in nonexercised horses (Table 1, Fig 1). The median half-life and the median mean residence time were significantly longer and median volume of distribution was significantly larger when detomidine was administered after exercise. Plasma clearance was not different between exercised and nonexercised horses. Discussion Intravenous detomidine produced lower peak plasma concentrations, was more widely distributed, and was eliminated at a slower rate when administered i.v. to horses immediately after maximal exercise compared to nonexercised horses. These results help to explain the increased dose requirement for sedatives in horses after maximal exercise (Hubbell et al. 1997a) and suggest that subsequent administrations, if necessary, may have a more prolonged effect. Lower peak plasma concentrations may provide a partial explanation for the previously established increased dose requirement for detomidine in horses post exercise (Hubbell et al. 1997a). While cardiac output was not measured prior to detomidine administration in nonexercised horses during this experiment, cardiac output was increased to 180% of pre-exercise values one minute after exercise in these same horses (Hubbell et al. 1997b). Cardiac output is a determinant of intravascular blood mixing with higher cardiac outputs producing lower initial plasma drug concentrations (Upton and Huang 1993), a principle utilised in measurement of cardiac output by indicator dilution (Hillidge and Lees 1975; Muir et al 1976). Drug delivery to target tissues is further influenced by changes in the distribution of cardiac output during and after exercise with reductions in the percentage of cardiac output delivered to target organs such as the brain to <30% of those at rest and blood flow increases (ml/kg bwt) to exercising muscles of up to 76-fold during maximal exercise (Parks and Manohar 1983). A direct relationship between increased cardiac output and increasing dose requirements for rapidly acting central nervous system depressant drugs (thiopental), attributed to increased intravascular drug mixing and tissue distribution has been described in man (Avram et al. 1993), providing further support for these concepts. Changes in muscle intracellular hydrogen ion concentration with exercise could also have contributed to the altered distribution and elimination. Detomidine is a lipophilic weak base with a pKa of 7.2 and a wide distribution (Salonen 1986). Decreases in intracellular pH during exercise increase the ionisation of weak bases leading to accumulation and trapping of weak bases in the cell (Gerweck 1998). Intracellular trapping could decrease peak plasma concentrations, increase the volume of distribution and delay elimination (Byrd et al. 1989; Gerweck 1998). The values reported here for median volume of distribution, mean residence time and median clearance are consistent with previous reports, particularly if the variations in dose are considered (Salonen et al. 1989; Elfenbein et al. 2009). The delayed elimination of detomidine seen in horses post exercise is probably the result of the increased volume of distribution and reductions of hepatic blood due to exercise-induced redistribution away from the viscera (liver) exacerbated by detomidine-induced reductions in cardiac output. Exercise decreases hepatic blood

flow as indexed by bromsulphalein clearance with the decreases extending into the recuperative period (Manohar et al. 1995; Dyke et al. 1998a,b). As previously reported, cardiac output decreased in horses recuperating from maximal exercise when detomidine was administered compared to nonmedicated, nonexercised horses (Hubbell et al. 1999). The delayed elimination of detomidine in horses after exercise has implications with regard to dose and dosing interval if subsequent administrations are required. Limitations of this study include the sampling sites used, the potential for unmeasured differences in haemoconcentration and plasma volume between groups, and the relatively narrow window used for sample collection. Plasma samples were collected via a catheter placed in the right atrium in the exercising horses and via a jugular catheter in the resting horses. Sampling site can have an effect on drug plasma concentrations if there is inadequate mixing or if one sampling site is immediately downstream from the eliminating organ (Upton 2000). The authors could find no reports documenting differences between jugular and right atrial plasma drug concentrations in horses. Further, liver blood flow is greatly reduced in exercising horses (Dyke et al. 1998a), limiting this potential effect. Differences due to sampling site, if any, would be reflected in the absolute plasma concentrations of drug but should not affect the rate of elimination. Packed cell volume and plasma volume were measured in exercised horses but not in resting horses. As previously reported, packed cell volume increased to levels 1.5 times baseline and remained elevated for 45 min and total plasma solids were significantly elevated for 15 min in the exercised group (Hubbell et al. 1999). The increases in packed cell volume and potential decreases in plasma volume (McKeever et al. 1993) would tend to increase the plasma detomidine concentrations unless detomidine partitions into red blood cells. This combination of factors could result in an underestimation of the effect of exercise on drug elimination by artificially elevating the plasma concentrations in exercising horses. The relatively narrow sampling interval was used to focus on the time period immediately after exercise during which exercised-induced alterations in blood flow, cardiac output, vascular tone and other physiological variables were expected to be maximally altered. A longer sampling period may have masked the effects we were seeking to identify when we were interested in the clinical effects of the drug. The potential clinical importance of this information is that increased doses of detomidine are required to achieve sedation in horses immediately after maximal exercise. This increased dose requirement could extend to other high cardiac output states (fever, stress) but the tissue distribution of cardiac output in those states has not been investigated. The duration of the attained post exercise sedation may be prolonged and subsequent doses should therefore be administered cautiously and to effect. Acknowledgements Supported in part by a grant from the Grayson Jockey Cub Research Foundation, Lexington, Kentucky. The authors thank D. Hall, J. Dutson, J. Gadowski, S. Schumacher, B. Badgely and W. Hsu for technical assistance, and Orion Farmos Corporation, Turku, Finland, for the detomidine and medetomidine reference compounds.

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Manufacturers addresses
1Sato

Treadmill, Uppsala, Sweden. Dickinson and Company, Parsippany, New Jersey, USA. Vascular Access Systems, Rutherford, New Jersey, USA. 4Agilent Technologies, Inc., Santa Clara, California, USA. 5InnaPhase Corporation, Philadelphia, Pennsylvania, USA.
2Becton 3Becton-Dickinson

Hubbell, J.A.E., Hinchcliff, K.W., Schmall, L.M., Muir, W.W., Robertson, J.T. and Sams, R.A. (1999) Cardiorespiratory and metabolic effects of xylazine, detomidine, and a combination of xylazine and acepromazine administered after exercise in horses. Am. J. vet. Res. 60, 1271-1279. Manohar, M., Goetz, T.E., Saupe, B., Hutchens, E. and Coney, E. (1995) Thyroid, renal, and splanchnic circulation in horses at rest and during short term exercise. Am. J. vet. Res. 56, 1356-1361. McKeever, K.H., Hinchcliff, K.W., Reed, S.M. and Robertson, J.T. (1993) Role of decreased plasma volume in hematocrit alterations during incremental treadmill exercise in horses. Am. J. Physiol. 265, R404-R408. Muir, W.W., Skarda, R.T. and Milne, D.W. (1976) Estimation of cardiac output in the horse by thermodilution techniques. Am. J. vet. Res. 37, 697-700. Pagliari, R. and Peyrin, L. (1995) Norepinephrine release in the rat frontal cortex under treadmill exercise: a study with microdialysis. J. appl. Physiol. 78, 2121-2130. Parks, C.M. and Manohar, M. (1983) Distribution of blood flow during moderate and strenuous exercise in ponies (Equus caballus). Am. J. vet. Res. 44, 1861-1866. Ramsay, E.C., Geiser, D., Carter, W. and Tobin, T. (2002) Serum concentrations and effects of detomidine delivered orally to horses in three different mediums. Vet. Anaesth. Analg. 29, 219-222. Salonen, J.S. (1986) Pharmacokinetics of detomidine. Acta vet. Scand. 82, 59-66. Salonen, J.S., Vaha-Vahe, T., Vainio, O. and Vakkuri, O. (1989) Single-dose pharmacokinetics of detomidine in the horse and cow. J. vet. Pharmacol. Therap. 12, 65-72. Snow, D.H., Harris, R.C., Macdonald, I.A. Forster, C.D. and Marlin, D.J. (1992) Effects of high-intensity exercise on plasma catecholamines in the Thoroughbred horse. Equine vet. J. 24, 462-467. Upton, R.N. (2000). Relationships between steady state blood concentrations and cardiac output during intravenous infusions. Biopharm. Drug. Dispos. 21, 69-76. Upton, R.N. and Huang, Y.F. (1993) Influence of cardiac output, injection time and injection volume on the initial mixing of drugs with venous blood after I.V. bolus administration to sheep. Br. J. Anaesth. 70, 333-338. Wagner, A.E., Muir, W.W. and Hinchcliff, KW. (1991) Cardiovascular effects of xylazine and detomidine in horses. Am. J. vet. Res. 52, 651-657.

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Author contributions All authors contributed to the initiation, conception, planning, execution and writing for this study. The pathology and statistics were by J.A.E.H. and R.A.S.