Influence of renal failure on the disposition of morphine, morphine-3-glucuronide and morphine-6-glucuronide in sheep during intravenous infusion with morphine.

The influence of experimentally induced renal failure on the disposition of morphine, morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) was examined in seven sheep infused intravenously with morphine for 6 hr. Between 5 and 6 hr, blood was collected from the aorta, pulmonary artery, hepatic, hepatic portal and renal veins and posterior vena cava. Additional samples from the aorta and urine were collected up to 144 hr. Morphine, M3G and M6G were determined in plasma and urine by high-performance liquid chromatography. Constant concentrations of morphine, but not of M3G and M6G, were achieved in plasma between 5 and 6 hr. Significant (P < .001) extraction of morphine by the liver (0.72 +/- 0.05) and kidney (0.42 +/- 0.15) occurred. Compared with sheep with normal kidneys (Milne et al., 1995), renal failure did not alter (P = .11) the mean total clearance of morphine (1.5 +/- 0.3 liters/min); clearance by the kidney was less (P < .001). However, a paired comparison using sheep common to this study and from the study when their kidneys were normal revealed a significant reduction in mean total clearance of 25%. The renal extraction of M3G and M6G and urinary recovery of the dose as summed morphine, M3G and M6G were reduced by renal failure. The kidney metabolized morphine to M3G. The data suggest that nonrenal elimination of M3G becomes more important during renal failure.

Lack of effect of gender and oral contraceptive steroids on the pharmacokinetics of (R)-ibuprofen in humans.

The effects of gender and oral contraceptive steroids on the pharmacokinetics of (R)-ibuprofen were studied in groups of healthy adult males, females and oral contraceptive steroid (OCS) using females. The values of AUC, CLpo, t1/2 and Vss, app did not differ significantly between the groups. Similarly, the percentage unbound of (R)-ibuprofen in pooled plasma from the three groups was not statistically different. Since chiral inversion is the major determinant of (R)-ibuprofen clearance in humans, it may be inferred from these data that gender and OCS have little or no effect on conversion of (R)-ibuprofen to the pharmacologically active S-enantiomer. Moreover, it is unlikely that hormonal factors influence the activity of the human hepatic long-chain fatty-acid:CoA ligase, the enzyme mediating the rate limiting step of (R)-ibuprofen inversion.

PIP: In Australia, clinical researchers studied the effects of gender and oral contraceptive (OC) steroids on the pharmacokinetics of (R)-ibuprofen in 8 healthy adult males (mean age = 21 years), adult females (24 years), and OC users (22 years). There were no statistically significant differences between males, females, and OC users for any pharmacokinetic parameter for (R)-ibuprofen. These parameters included areas under the plasma total concentration-time curve (AUC), maximal plasma concentration, time to maximal plasma concentration, half-life, CLpo, and apparent steady-state volumes of distribution. The AUC to the last data point observed for (S)-ibuprofen (derived from (R)-ibuprofen) was similar, suggesting that hormonal factors do not affect plasma clearance of (S)-ibuprofen. The average percentages unbound of (R)- and (S)-ibuprofen across the concentration was not statistically different between the groups: 1.82% and 2.84% for males, 1.83% and 3.01% for females, and 2.1% and 2.97% for OC users, respectively. The mean fraction unbound of (S)-ibuprofen was 53.6% greater than that of (R)-ibuprofen. Since chiral inversion may explain 62-92% of (R)-ibuprofen clearance in humans, these data may suggest that gender and OCs do not effect or have only a limited effect on the conversion of (R)-ibuprofen to the pharmacologically active S-enantiomer. These findings indicate that hormonal factors probably do not affect the activity of the human hepatic long-chain fatty-acid:CoA ligase, the enzyme mediating the rate limiting step of (R)-ibuprofen inversion.

Comparative disposition of morphine-3-glucuronide during separate intravenous infusions of morphine and morphine-3-glucuronide in sheep. Importance of the kidney.

The disposition of morphine-3-glucuronide (M3G) in sheep was compared during separate constant infusions of morphine and M3G. Five ewes received a 15-min loading dose, followed by a constant infusion of morphine sulfate (10 mg/hr) or M3G (4 mg/hr for 4 sheep, 7.5 mg/hr for 1 sheep) for a further 5.75 hr. During the 5th-6th hr of infusion, blood was collected simultaneously from the aorta, pulmonary artery, hepatic vein, hepatic portal vein, renal vein, and posterior vena cava. Additional samples were collected from the aorta from 0 to 5 hr and from 6 to 48 hr. Urine was collected via an indwelling catheter from 0 to 6 hr, with further free-flowing urine up to 48 hr. An HPLC assay was used to determine simultaneously morphine, M3G, and morphine-6-glucuronide (M6G) in plasma and urine. Constant concentrations of morphine, M3G, and M6G in plasma were achieved during the 5- to 6-hr period of infusion with morphine, as were the concentrations of M3G while M3G was infused. Regional net extraction ratios and total and regional clearances were calculated during the 5- to 6-hr period. After the infusions were ceased, there was prolonged elimination of M3G formed in situ from morphine compared to when infused as M3G. No morphine or M6G was detected in the plasma during and after infusion with M3G, nor were they found in urine collected up to 6 hr.(ABSTRACT TRUNCATED AT 250 WORDS)

Disposition of morphine and its 3- and 6-glucuronide metabolites during morphine infusion in the sheep.

A sheep preparation was used to examine the regional formation and extraction of morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G), relative to the regional extraction of morphine, at four morphine dose rates. On separate occasions, four ewes received a 15-min loading infusion of morphine sulfate, followed by a constant infusion at 2.5, 5, 10, or 20 mg/hr for an additional 5.75 hr. During the 5th to 6th hr of infusion, blood samples were collected simultaneously from the aorta, pulmonary artery, hepatic vein, hepatic portal and renal veins, posterior vena cava, and coronary and sagittal sinuses. Urine was collected for 48 hr. Morphine, M3G, and M6G in plasma and urine were determined simultaneously by HPLC. The blood/plasma concentration ratio (lambda) for morphine, M3G, and M6G was determined in spiked "blank" blood. Steady-state plasma concentrations were achieved during the sampling period, and dose-normalized concentrations were independent of the infusion rate. There was significant (p < 0.05) extraction (mean +/- SD) of morphine by the liver (0.676 +/- 0.014) and kidney (0.602 +/- 0.039), net extraction of M3G (0.106 +/- 0.046) and M6G (0.104 +/- 0.030) by the kidney, and net formation of M3G (-0.057 +/- 0.017) by the gut. The mean lambda for morphine, M3G, and M6G was 1.25 +/- 0.17, 0.80 +/- 0.03, and 0.82 +/- 0.09, respectively. The mean total body clearance of morphine with respect to blood was 1.58 +/- 0.27 liters/min. Mean (+/-SD) percentage urinary recoveries as morphine, M3G, and M6G were 14.7 +/- 8.5, 75.4 +/- 11.1, and 0.49 +/- 0.39, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Tissue distribution of bupivacaine enantiomers in sheep.

rac-Bupivacaine HCl was infused intravenously to constant arterial blood drug concentrations in sheep using a regimen of 4 mg/min for 15 min followed by 1 mg/min to 24 h. At 24 h, arterial blood was sampled, the animal was killed with a bolus of KCl solution, then rapidly dissected and samples were obtained from heart, brain, lung, kidney, liver, muscle, fat, gut, and rumen. Tissue:blood distribution coefficients for (+)-(R)-bupivacaine exceeded those of (-)-(S)-bupivacaine (P < 0.05) for heart, brain, lung, fat, gut, and rumen by an overall mean of 43%. Blood:plasma distribution coefficients of (-)-(S)-bupivacaine exceeded those of (+)-(R)-bupivacaine by a mean of 29% and this offset the tissue:blood distribution coefficients so that the previously significant enantioselective differences disappeared. It is concluded that although enantioselectivity of bupivacaine distribution is shown by the measured tissue:blood distribution coefficients, it is not shown when tissue:plasma water distribution coefficients are calculated, suggesting that there is no intrinsic difference between the bupivacaine enantiomers in tissue affinity. Sheep given fatal intravenous bolus doses of rac-bupivacaine had significantly greater concentrations of (+)-(R)-bupivacaine than (-)-(S)-bupivacaine in brain (P = 0.028) and ventricle (P = 0.036); these could augment the greater myocardial toxicity of this enantiomer found in vitro.

Failure of the nitrous oxide tissue equilibration method for the determination of brain and myocardial blood flow under controlled conditions.

1. Two adult merino ewes were prepared with intravascular cannule for sampling aortic root blood, sagittal sinus blood and coronary sinus blood. 2. One week after preparation the animals were anaesthetized then ventilated with a gas mixture containing 10% nitrous oxide (N2O) for 60 min. Serial measurements of brain and myocardial blood flow were made using the N2O tissue equilibration method of Kety and Schmidt. 3. N2O failed to achieve matching arteriovenous blood concentration equality and saturation of the relevant tissues. Valid use of the Kety-Schmidt method, therefore, could not be confirmed. 4. Because of the failure of the arteriovenous equilibration, serially determined brain and myocardial blood flows were found to decrease with time. 5. The use of this method in circumstances where tissue saturation with the indicator gas cannot be ascertained is arbitrary.

Physiological disposition of i.v. morphine in sheep.

In a crossover design study we have measured the total body and regional clearances of morphine. Thirteen experiments were performed in four conscious sheep that had been prepared previously with appropriate intravascular cannulae. Morphine (as sulphate pentahydrate) was infused i.v. at 2.5, 5, 10 and 20 mg h-1 to produce constant blood concentrations. Morphine (base) concentrations were measured in blood, urine and tissues with a specific HPLC method. The mean (SEM) total body clearance of morphine was 1.63 (0.21) litre min-1; this comprised 1.01 (0.10) litre min-1 clearance by the liver and 0.55 (0.06) litre min-1 by the kidneys. There was no evidence of dose-dependent clearance or significant extraction of morphine by the lungs, brain, heart, gut or hindquarters at any dose. The kidney clearance of morphine was greater than the 0.21 (0.06) litre min-1 renal clearance determined from the product of the mean total body clearance and the 12.3 (2.4%) of the administered dose recovered as unmetabolized morphine from 48 h urine collection (P less than 0.05). It was concluded that the liver and kidneys account for the majority of morphine clearance, and that the kidneys both excrete and metabolize morphine.

Cardiovascular effects and regional clearances of i.v. bupivacaine in sheep: enantiomeric analysis.

All currently available aminoacylaniline local anaesthetics, except lignocaine, contain a chiral centre but are used as racemates, a fact usually ignored in pharmacokinetic studies. This study reports the cardiovascular effects, and the regional and total body clearances of the enantiomers of bupivacaine determined at two steady state periods (3-4 h and 23-24 h) during continuous i.v. infusion to subtoxic concentrations in conscious sheep. Racemic (RS)-bupivacaine hydrochloride 1 mg min-1, was infused in five sheep that had been prepared at least 1 week previously with appropriate intravascular cannulae. The infusion of RS-bupivacaine produced constant arterial R(+)- and s(-)-bupivacaine concentrations of 0.20-0.68 mg litre-1 and 0.22-0.94 mg litre-1, respectively. This caused no appreciable cardiovascular effects. The hepatic clearance of R(+)- was greater than that of S(-)-bupivacaine (P less than 0.05) with mean (SD) clearance at the two respective time periods being 1.37 (0.78) and 1.47 (0.57) litre min-1 and 1.01 (0.72) and 1.29 (0.47) litre min-1. There was no significant clearance of either enantiomer by the lungs, brain, heart, gut, kidneys or hindquarters. It was concluded that, although the clearances of the enantiomers differed, the total body clearance of both enantiomers was accounted for by hepatic clearance exclusively. There was no evidence of time dependent kinetics.

Failure of the Kety-Schmidt nitrous oxide method for determination of myocardial blood flow.

1. The reliability of the Kety-Schmidt nitrous oxide (N2O) blood-tissue equilibration method was examined in 50 studies of myocardial blood flow in seven conscious, unrestrained sheep using a newly developed carefully validated gas chromatographic assay for N2O. 2. In 10 studies the arterial and coronary sinus N2O blood concentration-time curves converged as expected at the end of the 10 min sampling period. In 14 studies they crossed over, and in 26 studies, the curves failed to converge. 3. A survey of the literature revealed that such results have been encountered previously but have not been accorded particular significance. An ultimate matching equilibrium between arterial and venous blood N2O concentration-time curves is, however, fundamental to the validity of the method. 4. The results indicate that the use of the Kety-Schmidt method with N2O as the indicator gas is invalid as applied to the measurement of myocardial blood flow in this preparation.

Uptake and elution of chlormethiazole, meperidine, and minaxolone in the hindquarters of sheep: implications for clearance calculations.

Mass balance principles were used to describe the uptake and elution of chlormethiazole, meperidine, and minaxolone in the hindquarters of sheep. Sheep received a right atrial infusion of either chlormethiazole (3.71 mg/min) or meperidine (2.70 mg/min) for 180 min, or minaxolone (0.37 mg/min) for 120 min. Paired arterial and inferior vena cava (draining the hindquarters) blood samples were taken during and after the infusion. The mean and SD (n = 4) of the time-averaged extraction ratios across the hindquarters (determined from the relevant arterio-venous area under blood concentration--time curves) were 0.12 (0.10), 0.36 (0.13), and 0.27 (0.05) for chlormethiazole, meperidine, and minaxolone, respectively. The rank order of the rate of uptake of the drugs into the hindquarters was the same as the rank order of their lipophilicity, and uptake still continued when both the arterial and inferior vena cava drug concentrations were essentially constant. For chlormethiazole, meperidine, and minaxolone, 48% (44), 4% (6), and 35% (17), respectively, of the drug taken into the hindquarters eluted from the hindquarters after the infusion. Drug uptake and retention in extravisceral tissues, represented here by the hindquarters, can result in the mean total body drug clearance being overestimated when determined by traditional systemic pharmacokinetic methods.

The in vivo blood, fat and muscle concentrations of lignocaine and bupivacaine in the hindquarters of sheep.

1. A method was developed for sampling muscle and fat from the hindquarters of sheep undergoing spinal anaesthesia. The method was used to measure the concentrations of lignocaine and bupivacaine in the blood, muscle and fat of the hindquarters of sheep during and after 180 min constant-rate infusions of the drugs. 2. For both drugs the muscle drug concentrations were a relatively constant ratio of the simultaneous arterial blood drug concentrations during and after the infusion. 3. There was uptake of both lignocaine and bupivacaine into subcutaneous fat during the infusions. At the end of the infusion the ratio of the fat: arterial blood drug concentrations were 1.54 (SD = 0.57, n = 4) and 3.1 (SD = 1.4, n = 4) for lignocaine and bupivacaine, respectively. 4. The drug concentrations in fat declined relatively slowly after the infusion. The ratio of the fat: arterial blood drug concentrations 180 min after the end of the infusion was 21.5 (SD 4.0, n = 3) and for lignocaine, and 120 min after the end of the infusion was 9.54 (SD 5.2, n = 3) for bupivacaine. 5. It was concluded that the concentrations of lignocaine and bupivacaine in muscle were essentially in equilibrium with the arterial concentrations during and after the infusion. However, the concentrations of lignocaine and bupivacaine in fat were not in equilibrium with the arterial concentrations in the post-infusion period.

Improved method for morphine determination in biological fluids and tissues: rapid, sensitive and selective.

Morphine was assayed using a simple two step solvent extraction--acid back extraction sample preparation method, coupled with normal phase high-performance liquid chromatography (HPLC) and dual electrode coulometric detection. HPLC is performed with a 1.0 M Tris-methanol (5:95) mobile phase with subtle pH adjustments to separate morphine and internal standard from any interfering compounds. The use of normal phase HPLC (silica column) substantially reduces problems from interfering lipophilic substances sometimes encountered with reverse phase HPLC following solvent extraction and which would otherwise require more time-consuming sample preparation. Dual electrode detection further improves the selectivity for morphine and gives excellent sensitivity (0.5 ng mL-1), reproducibility and stability for automated sample injection. This method has proven suitable for pharmacokinetic studies of morphine.

Cardiovascular effects and regional clearances of intravenous ropivacaine in sheep.

The purpose of this study was to determine the cardiovascular effects and the total body and regional clearances of ropivacaine during its continuous intravenous infusion to subtoxic levels in five conscious unrestrained sheep that had been previously prepared with appropriate intravascular cannulas. Ropivacaine HCl.H2O, 1 mg/min, produced constant arterial blood concentrations which ranged from 0.70 to 1.84 mg/L. This caused no appreciable cardiovascular effects. The mean total body clearance (+/- SD) of ropivacaine was 1.00 +/- 0.27 L/min. There was significant clearance of ropivacaine by the liver (0.85 +/- 0.32 L/min), gut (0.09 +/- 0.07 L/min), and kidneys (0.04 +/- 0.03 L/min). There was no significant clearance of ropivacaine by the lungs, brain, heart, or hindquarters. It was concluded that the liver accounts for the majority of ropivacaine clearance.

Propofol: assay and regional mass balance in the sheep.

1. Pharmacokinetic data for propofol, a new intravenous anaesthetic agent, indicate that there may be extensive extrahepatic clearance. This was investigated during intravenous infusions of propofol in adult merino ewes with chronic intravascular cannulae using a newly developed simple and rapid assay for propofol in blood and other biological samples. 2. The assay was based on organic solvent extraction of pH 4.5 buffered blood, urine or tissue homogenate, followed by reverse-phase h.p.l.c. with fluorescence detection. 3. A mean total body clearance of propofol of 3.15 l/min, (SD 0.87 l/min; n = 8) was found, consistent with a high hepatic extraction ratio (overall mean 0.87, SD 0.19; n = 8) and clearance (overall mean 1.12, SD 0.25 l/min; n = 7). The difference between total and hepatic clearances consisted principally of pulmonary clearance, but its extent was variable. 4. Other regional pharmacokinetic data were consistent with propofol distribution into muscle and brain tissues and propofol 'production' by the kidney, probably from a propofol metabolite formed elsewhere. 5. If these data are confirmed in humans then clinical pharmacokinetic data so far derived from peripheral venous blood sampling will require re-evaluation.

Myocardial and cerebral drug concentrations and the mechanisms of death after fatal intravenous doses of lidocaine, bupivacaine, and ropivacaine in the sheep.

This paper reports the cardiovascular effects of intentionally toxic intravenous doses of lidocaine, bupivacaine, and ropivacaine and the mechanisms of death. Fatal doses of lidocaine, bupivacaine, and ropivacaine were established in sheep treated with successive daily dose increments of each drug. The mean fatal dose of lidocaine (+/- SD) was 1450 +/- 191 mg (30.8 +/- 5.8 mg/kg), that of bupivacaine was 156 +/- 31 mg (3.7 +/- 1.1 mg/kg), and that of ropivacaine was 325 +/- 108 mg (7.3 +/- 1.0 mg/kg); thus the ratio of fatal doses was approximately 9:1:2. In four out of four lidocaine-treated animals, respiratory depression with bradycardia and hypotension without arrhythmias were the causes of death. Three out of four bupivacaine-treated animals died after the sudden onset of ventricular tachycardia/fibrillation without hypoxia or acidosis; the fourth died in a similar manner to the lidocaine-treated animals. Three out of five animals given ropivacaine died in a manner resembling the fatal effects of lidocaine-treated animals, but unlike the lidocaine-treated animals, in all three sheep there were also periods of ventricular arrhythmias. The remaining two ropivacaine-treated sheep died as a result of the sudden onset of ventricular tachycardia/fibrillation. The mean percentages of the fatal dose found in the myocardium was 2.8 +/- 0.7 for lidocaine-treated animals, 3.3 +/- 0.9 for bupivacaine-treated animals, and 2.2 +/- 1.4 for ropivacaine-treated animals; the corresponding percentages in whole brain were, respectively, 0.71 +/- 0.01, 0.71 +/- 0.21, and 0.89 +/- 0.27.

The uptake and elution of lignocaine and procainamide in the hindquarters of the sheep described using mass balance principles.

Mass balance principles were used to describe the uptake and elution of lignocaine (lidocaine) and procainamide in the hindquarters of the sheep. Each of four sheep received a right atrial infusion of either lignocaine.HCl (2.7 mg/min) or procainamide.HCl (5.5 mg/min) for 180 min. Paired arterial and inferior vena cava (draining the hindquarters) blood samples were taken at 20-min intervals during the infusion and for 180 min after the infusion. Lignocaine and procainamide mean total body clearances were 2.9 L/min (SD 1.1) and 1.3 L/min (SD 0.2), respectively. An index of the uptake and elution of these drugs in the hindquarters was estimated from the net drug mass per unit hindquarter blood flow; indirect evidence suggested that hindquarter blood flow was constant. All the net mass/flow of procainamide that was taken into the hindquarters during the infusion also eluted after the infusion, demonstrating reversible distribution into the tissues. However, uptake of procainamide was still occurring when blood concentrations were constant, indicating that the concentrations of procainamide in the hindquarters were not in equilibrium with the inferior vena cava concentrations. Lignocaine did not reach constant blood concentrations during the infusion and showed no tendency to reach arterio-venous equilibration; an arterio-venous difference of 22% (SD 5%) across the hindquarters was measured during the last 60 min of the infusion. By 180 min after the lignocaine infusions, 79% (SD 8%) of the lignocaine net mass/flow had not eluted from the hindquarters when arterial and venous lignocaine concentrations were not significantly different. This drug could remain uneluted due to metabolism and/or avid tissue binding, and presents difficulties in the interpretation of pharmacokinetic data whether based on arterial or venous blood sampling.

Sensitive gas liquid chromatography method for the determination of fentanyl concentrations in blood.

A method for the determination of fentanyl blood concentrations using gas liquid chromatography coupled to a nitrogen phosphorus detector (NPD) is presented. A highly inert fused-silica, megabore column coated with a methyl silicone stationary phase was used for the analysis. The mean coefficient of variation for the range of fentanyl concentrations tested (0.25-10 ng/ml) was 4.65%, ranging from 0.85% at 10 ng/ml to 10.8% at 0.25 ng/ml. The assay was used to quantify blood fentanyl concentrations collected from a 56-year-old woman who was administered fentanyl postoperatively via a patient-controlled on-demand analgesic computer (ODAC). The mean hourly fentanyl dose rate over the 44 hr study period was 41.8 micrograms/hr (range 20-120). The sixfold variation in hourly dose rate was not mirrored by similar fluctuations in the fentanyl blood concentration (mean 0.45 ng/ml, range 0.3-0.7 ng/ml). The patient thus titrated herself to a perceived minimum effective concentration (MEC) of fentanyl.

The effects of general anaesthesia on tocainide clearance in the sheep.

The haemodynamic effects and regional clearances of tocainide were investigated in sheep with chronic intravascular cannulae to measure blood flow through, and drug extraction by, lungs, kidneys, liver and gut. 2. Tocainide, at arterial blood concentrations in the therapeutic range, caused no haemodynamic effects and was significantly extracted only by the liver. 3. In the presence of general anaesthesia with halothane, the mean hepatic blood flow and tocainide extraction ratio were each reduced by approximately 25% so that the mean hepatic clearance and intrinsic clearance of tocainide each were reduced by approximately 50%. Thus arterial blood tocainide concentrations were increased by 50%. 4. While the clinical implications of this interaction are unclear because of insufficient information about the margin of safety of tocainide, the pharmacological implications are plain. Because general anaesthesia may alter the relationship between dose and blood drug concentrations, pharmacokinetic and pharmacodynamic data should not be interchanged between awake and anaesthetized subjects.

Effects of combined extradural blockade and general anaesthesia on indocyanine green clearance and halothane metabolism.

Cardiovascular effects, indocyanine green clearance and the reductive metabolism of halothane were compared in 10 patients having peripheral surgery who received either general anaesthesia or general anaesthesia plus extradural blockade. Bupivacaine or saline was given extradurally 1 h before anaesthesia with nitrous oxide:oxygen (70:30) and halothane (end-tidal concentration 0.35%). Indocyanine green clearance was measured before (stage I) and after (stage II) bupivacaine or saline, during anaesthesia before surgery (stage III), and during surgery (stage IV). Reductive metabolism of halothane was assessed by measurement of exhaled 2-chloro-1,1,1-trifluoroethane. In the general anaesthesia plus extradural blockade group, arterial pressure was decreased to 58% (SD = 6) of stage I values over stages II-IV, whereas no changes were observed in the general anaesthesia group. There was no significant difference in indocyanine green clearance or the reductive metabolism of halothane, despite the occurrence of significant decreases in arterial pressure.