Population distributions of phenylbutazone and oxyphenbutazone after oral and i.v. dosing in horses.

Experiments to determine the residual plasma concentrations of phenylbutazone and its metabolites found in horses racing on a 'no-race day medication' or 24-h rule were carried out. One dosing schedule (oral-i.v.) consisted of 8.8 mg/kg (4 g/1000 lbs) orally for 3 days, followed by 4.4 mg/kg (2 g/1000 lbs) intravenously on day 4. A second schedule consisted of 4.4 mg/kg i.v. for 4 days. The experiments were carried out in Thoroughbred and Standardbred horses at pasture, half-bred horses at pasture, and in Thoroughbred horses in training. After administering the i.v. schedule for 4 days to Thoroughbred and Standardbred horses at pasture, the mean plasma concentrations of phenylbutazone increased from 0.77 microgram/ml on day 2 to 2.5 micrograms/ml on day 5. The shape of the frequency distribution of these populations was log-normal. These data are consistent with one horse in 1,000 yielding a plasma level of 8.07 micrograms/ml on day 5. After administration of the oral-i.v. schedule to Thoroughbred and Standardbred horses at pasture, the mean plasma concentrations of phenylbutazone were 3.4 micrograms/ml on day 2 and 3.5 micrograms/ml on day 5. The range on day 5 was from 1.4 to 8.98 micrograms/ml and the frequency distribution was log-normal. These data are consistent with one horse in 1000 having a plasma level of 15.8 micrograms/ml on day 5. In a final experiment, the oral dosing schedule was administered to 62 Thoroughbred horses in training. Plasma concentrations on day 5 in these horses averaged 5.3 micrograms/ml. The range was from 1.3 to 13.6 micrograms/ml and the frequency distribution was log-normal. Statistical projection of these values suggests that following this oral dosing schedule in racing horses about one horse in 1000 will yield a plasma level of 23.5 micrograms/ml of phenylbutazone 24 h after the last dose.

The effects of naloxone on endotoxic and hemorrhagic shock in horses.

The effects of naloxone on the cardiovascular, hematologic and metabolic derangements associated with endotoxic and hemorrhagic shock were studied in unanesthetized horses. In the first of 3 experiments blood glucose and lactate levels, hematocrit, white, red and differential white cell counts, rectal temperature and clinical signs were obtained before and after endotoxin (10 micrograms/Kg) administration in 5 horses. In the second experiment, two groups of 3 horses received either intravenous naloxone (0.04 mg/Kg) or saline, 7 minutes prior to endotoxin. In a third experiment two groups of 4 horses received either saline or naloxone (0.20 mg/Kg) immediately following acute hemorrhage. In the second and third experiments, pulse, mean arterial and right ventricular pressures, and heart rate were also observed. Endotoxin and acute hemorrhage produced hypothermia, leukopenia, lymphopenia, neutropenia, elevations in hematocrit, blood glucose and blood lactate, and clinical signs of shock. Naloxone (0.040 mg/Kg IV) significantly lowered endotoxin-induced increases in right ventricular pressure and heart rate, and at a higher dose (0.20 mg/Kg) antagonized the decrease in pulse and heart rate, and tachycardia observed after acute hemorrhage. These results suggest endogenous opioids are involved in the pathogenesis of shock. Naloxone appeared to attenuate some of the cardiovascular responses associated with shock and thus may be of therapeutic value in shock management.

Pharmacokinetics and protein binding of morphine in horses.

Morphine could be detected in horses dosed with 0.1 mg of drug/kg of body weight for up to 48 hours in blood and 144 hours in urine. This dose of morphine elicited no observable effects and is a suggested analgesic dose. Computer analysis revealed that a 3-compartment open system was the best fitting model with a serum half life (t1/2(beta)) of 87.9 minutes and a urine t1/2(beta) of 101.1 minutes. Binding to equine serum proteins was linear over a drug concentration range of 3.88 X 10(-5)M to 3.50 X 10(-8)M and averaged 31.6%. In RBC-partitioning experiments, 78.1% of the drug was found in the plasma fraction. The data indicated that a horse should not be given morphine closer than 1 week before a race.

The pharmacology of furosemide in the horse. V. Pharmacokinetics and blood levels of furosemide after intravenous administration.

Studies were undertaken to determine blood levels of furosemide in horses after 0.5- and 1.0-mg/kg doses administered iv. Analyses indicated that the pharmacokinetic parameters were dose independent and best described by a three-compartment open model. The alpha-, beta-, and gamma-phase half-lives of 5.6, 22.3, and 158.5 min, respectively, were observed after the 0.5-mg/kg dose. Similarly, the respective half-lives after the 1.0-mg/kg dose were 5.8, 24.1, and 177.2 min. After a 0.5-mg/kg dose of furosemide, population frequency distributions were evaluated at 1 hr and 4 hr post-drug administration. At 1 hr after dosing, the blood levels of furosemide in 30 horses were normally distributed. The mean plasma level was 97.9 ng/ml with a range of 41.9 ng/ml to 155.8 ng/ml and a SD of 25.0 ng/ml. Analyses of blood levels of furosemide in 47 horses at 4 hr after drug administration indicated that the population distribution was better fit by a normal curve after log transformation of the values. The mean plasma level at 4 hr post-dosing was 9.6 ng/ml with a range of 4.0 ng/ml to 19.4 ng/ml and a SD of 3.1 ng/ml. Based on this population distribution of the plasma levels, the probability of finding furosemide plasma concentrations above 24.6 ng/ml at 4 hr after anti-epistaxis dose was estimated at less than 1 in 1000.

Effects of enkephalins versus opiates on locomotor activity of the horse.

The enkephalins are small, pentapeptide neurotransmitter molecules which have reportedly been used in racing horses. In our experiments, D-Ala2-Metenkephalinamide and leucine enkephalin were administered to horses intravenously (IV) and intracisternally (IC). Leucine enkephalin had little effect on locomotor activity by either route at doses of 0.01 mg/Kg or less. Methionine enkephalinamide, an enzyme resistant enkephalin analog, had no significant effect when given IV (0.002 and 0.008 mg/kg). Other experiments involving intracisternal dosing with this long acting form at higher levels (0.005-.011 mg/Kg), elicited an initial increase in locomotor activity, a rise in temperature, a marked increase in blood pressure, hyperventilation, the appearance of a rapid eye blinking reflex, lack of coordination and quivering. In contrast, dosing with fentanyl either IV (0.01) mg/Kg) or Ic (0.0002 mg/Kg) produced a tenfold increase in locomotor activity without accompanying adverse clinical symptoms. The data suggest that very large doses of IV administered enkephalins or their analogs may be necessary to increase locomotor activity but such doses may also elicit a number of less desirable side effect.

Morphine glucuronide hydrolysis: superiority of beta-glucuronidase from Patella vulgata.

beta-Glucuronidase from Patella vulgata, Helix aspersa, Helix pomatia, and bovine liver were evaluated for usefulness in routine hydrolysis of drug-glucuronic acid conjugates from equine urine samples. Factors affecting the reaction rate (enzyme concentration, ligand concentration, temperature, and pH) were optimized. A 3-h incubation at 65 degrees C with 5000 U of beta-glucuronidase from P. vulgata per milliliter of urine resulted in complete hydrolysis of all morphine glucuronide in the urine samples. Not only was the enzyme preparation from P. vulgata the most cost-effective beta-glucuronidase source studied, but also its thermal stability is such that it can be used at a temperature high enough to substantially shorten the incubation interval. Preliminary work on other drugs that form glucuronide conjugates indicates that this same procedure is similarly superior for use in their hydrolysis.

Furosemide, Patella vulgata beta-glucuronidase and drug analysis: conditions for enhancement of the TLC detection of apomorphine, butorphanol, hydromorphone, nalbuphine, oxymorphone and pentazocine in equine urine.

We have investigated the action of five sources of beta-glucuronidase enzymes on the hydrolysis of glucuronides of apomorphine, butorphanol, hydromorphone, nalbuphine, oxymorphone and pentazocine in equine urine. For all glucuronides tested, Patella vulgata beta-glucuronidase yielded the largest thin layer chromatographic (TLC) spots. For oxymorphone, P. vulgata was the only treatment to yield detectable TLC spots under test parameters. For these six drugs, TLC spot size and chromatographic quality were compared between control horses and horses pretreated with furosemide four hours earlier. Furosemide pretreatment produced a statistically significant increase in spot size and was found to enhance chromatogram quality. These findings support previous suggestions that P. vulgata is a superior drug-glucuronide hydrolyzing enzyme. They also support earlier reports that administration of furosemide at four hours pre-race is unlikely to result in significant interference with routine drug testing procedures.