Liquid-phase microextraction based on carrier mediated transport combined with liquid chromatography-mass spectrometry. New concept for the determination of polar drugs in a single drop of human plasma.

Recently, we demonstrated for the first time liquid-phase microextraction (LPME) of polar drugs based on carrier mediated transport. In this new extraction technique, selected analytes were extracted as ion-pairs from small volumes of biological samples, through a thin layer of a water immiscible organic solvent immobilised in the pores of a porous hollow fibre (liquid membrane), and into a microl volume of an acidic aqueous acceptor solution placed inside the lumen of the hollow fibre. In the current paper, this new extraction technique was combined with liquid chromatography-mass spectrometry (LC-MS) for the first time. Carrier mediated LPME was evaluated for several new model drugs (0.01

[Toxicologic analysis of some adrenergic-beta blockers in the diagnosis of intoxications]. / Analiza toksykologiczna wybranych beta-adrenolityków w diagnostyce zatruc.

The study aimed at finding effective techniques of qualitative and quantitative analysis of selected beta-adrenergic blockers, useful both for monitoring of therapy and for thanatological diagnosis of intoxications. The studies took advantage of gas chromatography (GLC) and high performance liquid chromatography (HPLC). For isolation of studied compounds from biological material, classical and solid phase extraction procedures (SPE) Extrelut-20 (Merck), Abselut Nexus (Varian), STRATA C--18 E (Phenomenex) were used. The program included the analysis of most frequently applied derivatives: Acebutolol, Atenolol, Bunitrolol, Bupranolol, Labetolol, Metipranolol, Metoprolol, Oxprenolol, Practolol, Propranolol.

Quantitative determination of clenbuterol enantiomers in human plasma by high-performance liquid chromatography using the macrocyclic antibiotic chiral stationary phase teicoplanin.

We report a method for the high-performance liquid chromatographic (HPLC) chiral separation of racemic clenbuterol in human plasma. Human plasma was spiked with stock solutions of clenbuterol hydrochloride and practolol as the internal standard. Following a liquid-liquid extraction procedure with 10% (+/-)-2-butanol/isopropyl ether under alkaline conditions, the dried samples were reconstituted in methanol and chromatographed using a macrocyclic antibiotic chiral stationary phase (CSP) known as Chirobiotic T(trade mark) (teicoplanin). The mobile phase composition was methanol:acetonitrile (70:30, v/v), containing 0.3% (v/v) acetic acid and 0.2% (v/v) triethylamine. The resulting chromatogram achieved baseline separation for the clenbuterol enantiomers. Calibration curves (peak area ratio vs plasma concentration, n = 10) were constructed for the (-)-R-and (+)-S-clenbuterol enantiomers with a plasma concentration range of 0. 25-10 microM. The correlation coefficient (r) range was 0.99988-0. 99999 (mean = 0.99999). The lowest concentration measured was 0.25 microM. Inter- and intra-assay variation was determined for the lowest, medium and highest plasma concentration (0.25, 2 and 10 microM) by calculating the analytical recoveries with a range of 96-104%. The percentage recoveries for the clenbuterol enantiomers were 88.4-102% over the concentration range used. Detailed methodology is presented.

Curve-fitting in pharmacokinetics--a comparison between gamma- and biexponential fits.

Seven sets of plasma concentration-time data were fitted to both a conventional biexponential equation and a gamma equation. The values of clearance (CL) and mean residence time (MRT) were calculated from the fitted parameters and compared with the values calculated by the trapezoid rule. Both the biexponential and gamma equations provided adequate fits to the data. The values of CL and MRT calculated from the biexponential fits correlated very closely with the values calculated by the trapezoid rule, but there were large discrepancies between the values calculated from the gamma fits and the trapezoid rule. The biexponential model appears to be less sensitive to scatter in the data.

Effects of beta-adrenoceptor stimulation on rectosigmoid motility in man.

Effects of selective beta-adrenoceptor agonists on rectosigmoid motility during prolonged rectal distension were studied in 12 healthy volunteers in a double-blind, randomized fashion. Continuous distension was performed with a balloon in the proximal part of the rectum. Pressure was recorded by this balloon and by a catheter in the sigmoid. Contractile activity was quantified for three consecutive periods of 25 min. On separate days prenalterol (beta-1 agonist), terbutaline (beta-2 agonist), and placebo, respectively, were administered intravenously preceded by a control period. Terbutaline, 0.5 mg intravenously, was followed by a significant decrease of sigmoid motility from 4.3 +/- 1.5 (SEM) kPa X min to 2.9 +/- 1.0 kPa X min (P less than 0.01) and of rectal motility from 4.3 +/- 1.3 to 2.4 +/- 0.7 kPa X min (P less than 0.05). After placebo a slight, but not significant, increase of contractile activity was seen compared to the initial control period. The effects of prenalterol, 1.0 and 4.0 mg intravenously, on motility did not differ from that of placebo infusion. Both drugs caused a dose-dependent increase of systolic blood pressure and of heart rate. The study shows that beta-2-adrenoceptor stimulation decreases rectosigmoid colonic pressure in man, while effects of beta-1 stimulation on motility index do not differ from that of placebo.

Long-term hemodynamic effects of prenalterol in patients with severe congestive heart failure.

In a controlled, randomized, double-blind study we investigated the long-term effects of the beta 1-adrenoceptor agonist prenalterol in 16 patients with severe congestive heart failure (NYHA class III or IV). Previous to and 1 week, 3 months, and 6 months after continuous oral intake of 40 to 120 mg prenalterol a day, catheterization of the right heart combined with an ergometer test was carried out; M mode and two dimensional echocardiograms as well as systolic time intervals were also recorded. With prenalterol the heart rate increased within 1 week from 81 +/- 7 to 90 +/- 7 beats/min (mean +/- SD) (p less than .05) and remained increased after 3 months (93 +/- 9 beats/min, p less than .01) and 6 months (91 +/- 6 beats/min, p less than .05). After 1 week the cardiac index rose from 2.7 +/- 0.7 to 3.3 +/- 0.7 l/min/m2 (p less than .01), and after 3 and 6 months it fell again to 3.0 +/- 0.9 l/min/m2 and 2.9 +/- 0.7 l/min/m2, respectively. In the ergometer test the improvement in performance was not significant. The mean velocity of circumferential fiber shortening initially increased from 0.58 +/- 0.20 to 0.79 +/- 0.28 circumferences/sec (p less than .01), but dropped after 3 months to 0.62 +/- 0.31 circumferences/sec. The ejection fraction determined from the two-dimensional echocardiogram rose after 1 week from 20 +/- 10 to 27 +/- 12% (p less than .05), but decreased again after 3 months (23 +/- 11%) and 6 months (20 +/- 10%).(ABSTRACT TRUNCATED AT 250 WORDS)

Inotropic effect of prenalterol in amitriptyline poisoning.

A case of severe amitriptyline poisoning with grade IV coma, seizures, bradycardia and hypotension who did not respond to dopamine was successfully treated with prenalterol, a new cardioselective beta-agonist. The case is discussed with respect to plasma concentrations of dopamine, prenalterol and amitriptyline. Prenalterol, hydrocortisone and insulin may be useful as inotropic agents in tricyclic poisoning where dopamine fails to provide an adequate response.

Prenalterol as long-term therapy for chronic congestive heart failure. A randomized cross-over trial.

Ten patients with severe chronic congestive heart failure (CHF) due to ischaemic heart disease treated with digitalis and diuretics were randomly allocated to oral treatment with prenalterol (100-200 mg daily in addition to their basal treatment) or to intensified treatment with diuretics in a cross-over trial. A wash-out period of 1-4 weeks was allowed between the two modes of treatment. Most of the patients demonstrated subjective improvement during prenalterol therapy, but this improvement could not be verified objectively by exercise test, echocardiography, chest X-ray or weight measurements. No serious side-effects of either mode of treatment were observed. Heart rate was significantly lower during exercise when the patients were treated with prenalterol than during the control periods or during intensified conventional treatment, indicating that prenalterol acts as a beta-adrenergic receptor blocker during exercise in this patient group. The results indicate that prenalterol is a partial beta-receptor agonist without superior beneficial effects compared to those of intensified conventional treatment in patients with chronic, severe CHF.

A dose response study with oral prenalterol in patients with chronic congestive cardiac failure.

Prenalterol is an orally active cardioselective beta agonist, with a long half-life. Previous studies have confirmed its inotropic activity following intravenous infusion in patients with heart failure. It has little chronotropic activity and no significant arrhythmogenicity. We have studied the response to sustained-release oral prenalterol given over four weeks at doses of 20, 40, 100, and 200 mg daily in 10 patients with New York Heart Association class II and III heart failure due to ischemic heart disease. All were in sinus rhythm and already receiving diuretics and digoxin. The drug was well tolerated and without side effects. Nine patients showed a dose-related improvement in their exercise tolerance as measured on the treadmill, up to a dose of 100 mg daily, with a significant increase in estimated oxygen uptake. There was a dose-related reduction in maximum heart rate, systolic blood pressure, and rate-pressure product during exercise, which is suggestive of a reduction in myocardial oxygen consumption. We conclude that prenalterol improves exercise tolerance without any significant cardiovascular or other side effects, and produces a clinically relevant and sustained improvement in patients with chronic heart failure. M-mode echocardiographic measurements of left ventricular dimension and function at rest did not show any change during the study.

Cardioselectivity of prenalterol and isoproterenol.

We examined the hemodynamic effects and kinetics of prenalterol, a new beta-adrenoceptor agonist, in 10 normal subjects. There is some doubt whether prenalterol is selective for beta 1 adrenoceptors in animals; therefore, we also compared its cardioselectivity with that of the nonselective agonist, isoproterenol, with respect to heart rate (HR) and blood pressure (BP) responses after inhibition of cardiovascular reflexes with atropine, clonidine, and phentolamine. After intravenous (2.5 mg) and oral (10 mg and 100 mg) dosing, t 1/2 beta was 2 to 3 hr. Oral bioavailability averaged 33% and was independent of dose. Oral prenalterol, 10 mg and 100 mg, increased resting HR, systolic BP, and cardiac index by up to 27% but had no significant effects during graded exercise. Prenalterol infusions were calculated to attain steady-state plasma concentrations of 10, 20, and 40 ng/ml. HR and BP effects of the levels (10.8, 23.6, and 47.4 ng/ml) were compared with those of 0.5, 1.5, and 2.5 micrograms isoproterenol. Before autonomic block, prenalterol increased HR by 10 bpm at the highest dose and mean arterial pressure (MAP) by 10 mm Hg. In contrast, HR rose and MAP fell after isoproterenol. After block, at the highest doses of prenalterol and isoproterenol, there was an average rise in HR of 42 and 27 bpm; BP was almost maintained after the former but fell by 33 mm Hg after the latter. Prenalterol is an inotropic drug that has the effects of a full cardioselective beta-adrenoceptor agonist. Its inotropic effects are evident at doses that have little effect on HR because of the modifying effect of cardiovascular reflexes. The hemodynamic effects are most obvious at rest when sympathetic tone is low.

Electrophysiological effects of prenalterol on the cardiac conduction system.

The electrophysiological effects of prenalterol, a new beta-receptor agonist, in a dose of 100 micrograms/kg body weight given intravenously for 5 min, were studied in 13 patients with signs of sinus node dysfunction and/or conduction defects within or distal to the atrioventricular (AV) node. In nine patients with signs of sinus node dysfunction a significant reduction was found in corrected sinus node recovery time, on an average by 1955 +/- 640 ms (-61%, P less than 0.05) and in atrial refractoriness, by 61 +/- 21 ms (-20%, P less than 0.05). Similar but insignificant changes were also seen in the four patients with normal sinus node function. In eight patients with AV nodal dysfunction, a significant increase was found in the Wenckebach point, by 51 +/- 10 b.p.m. (+52%. P less than 0.01) and a decrease in the AH interval, by 23 +/- 9 ms (-14%, P less than 0.05). AV nodal refractoriness tended to decrease by 115 +/- 58 ms (-24%, NS). Similar changes were found in the five patients with normal AV conduction. Heart rate increased in all 13 patients, on an average by 28 +/- 5 b.p.m. (+44%, P less than 0.001) and systolic blood pressure by 18 +/- 8 mmHg (+13% P less than 0.01). In conclusion, prenalterol increased sinus node automaticity and atrial and AV nodal conductivity, but did not improve infranodal conduction. Thus, the drug might be useful in the treatment of patients with sinus node dysfunction as well as in patients with spontaneous or induced atrioventricular conduction abnormalities.

Haemodynamic and cardiac metabolic effects of the new beta 1 agonist prenalterol in patients with cardiac failure.

Prenalterol, a beta 1 selective agonist, exerts a positive inotropic action in animal studies as well as in human volunteers and is effective when administered orally. To assess its immediate haemodynamic and myocardial metabolic effects, we studied the response to prenalterol (50 and 100 micrograms kg-1 given intravenously by cardiac catheterization) in 15 patients with congestive heart failure secondary to coronary artery disease or non-ischaemic cardiomyopathy. At peak effect, cardiac index increased from 2.6 +/- 0.5 to 3.2 +/- 0.8 l min-1 m2 (mean +/- S.D.) (P less than 0.001); peak rate of left ventricular pressure development rose from 963 +/- 242 to 1335 +/- 411 mmHg s-1 (P less than 0.001); left ventricular end-diastolic pressure fell from 25 +/- 6 to 17 +/- 7 mgHg (P less than 0.001); coronary sinus blood flow increased from 113 +/- 39 to 148 +/- 55 ml min-1 (P less than 0.01); myocardial oxygen consumption was augmented from 12.7 +/- 3.9 to 16.4 +/- 5.8 ml min-1 (P less than 0.001); and heart rate increased slightly (from 76 +/- 12 to 86 +/- 14 beats min-1; (P less than 0.05)). No significant changes occurred in left ventricular systolic pressure, stroke volume index, myocardial lactate extraction rate and myocardial arteriovenous oxygen difference, and no patients developed angina, ECG changes or ventricular arrhythmias. Infusion of prenalterol effectively improved haemodynamic function and cardiac metabolism in cardiomyopathy. Therefore this agent deserves further investigation to evaluate its possible role for the long-term therapy of patients with chronic heart failure.

Ventilatory and haemodynamic effects of prenalterol and terbutaline in asthmatic patients.

In 8 asthmatic patients a comparative study was performed of the ventilatory and haemodynamic effects of the beta 1-receptor stimulator prenalterol and the beta 2-receptor stimulator terbutaline infused in increasing doses after a placebo. Terbutaline caused a dose-dependent decrease in diastolic blood-pressure (BP) and an increase in systolic BP and heart-rate (HR), while mean arterial pressure (MAP) did not change. Prenalterol produced a dose-dependent increase in MAP and systolic BP, while diastolic BP was unaffected. HR was increased only by the largest dose of prenalterol. The haemodynamic effects of the terbutaline infusion can be explained by a reflex response to the vasodilatation induced by stimulation of the vascular beta 2-receptors, while the effects of prenalterol can mainly be accounted for by a direct action on beta 1-receptors in the heart. These observations show that the cardiac side-effects of beta 2-agonists cannot be avoided by producing more selective agonists. Terbutaline caused a dose-dependent increase in the ventilatory indices. Prenalterol in larger doses caused a limited but significant increase in the ventilatory indices, comparable to the decrease in ventilation caused by the beta 1-selective blocker metoprolol. These findings suggest that the ventilatory effects of metoprolol and prenalterol are mediated via beta 1-receptors in the airways, which apparently play a functional role in asthma.

Assay of prenalterol in plasma and urine by gas chromatography-mass spectrometry.

A sensitive and selective method for the quantitative analysis of prenalterol in plasma and urine is described. Prenalterol and the internal standard, deuterated prenalterol, are extracted with diethyl ether at pH 9.9 under salting-out conditions. Derivatization is performed by means of pentafluoropropionic anhydride in toluene before separation in the gas chromatograph. Detection and quantification of the triacyl derivatives are done by mass spectrometry in the selected ion monitoring mode. The method allows determination of concentrations down to 5 nmol/l (1 ng/ml) in 1 ml of sample, with a relative standard deviation below 10%.

Inotropic, chronotropic and dromotropic effects of prenalterol, a new cardiostimulant drug.

Prenalterol (H 133/22) is an adrenergic beta-receptor stimulator which unlike isoproterenol is both orally active and has a long duration of action. The inotropic, chronotropic and dromotropic effects of prenalterol were investigated in pentobarbital anaesthetized dogs. Prenalterol was found to increase left ventricular maximum dP/dt in a dose-dependent manner up to a dose level of 50 micrograms/kg. Additional doses resulted in only small further increases in cardiac contractility. Electrophysiological studies were done to compare the effects of prenalterol and isoproterenol on sinus node and ventricular pacemaker function. Complete AV block was produced by electrocauterization of the His bundle. Prenalterol accelerated both ventricular and sinus node pacemakers at doses up to 50 micrograms/kg. The increase in ventricular rate was greater than the increase in sinus rate following supramaximal inotropic doses of prenalterol yet no ventricular extrasystoles were ever observed. Similar acceleration of the ventricular and sinus node pacemakers were observed by infusion of isoproterenol. Prenalterol and isoproterenol accelerated AV nodal conduction and ventricular conduction but had little effect upon His-Purkinje conduction. Sinus node reset time was abbreviated as was the functional and effective refractory periods of the atrium and AV nodes.

Metabolic effects of prenalterol in diabetic patients.

In 16 non-diabetic and 16 diabetic patients prenalterol, dobutamine and dopamine infusions (5 micrograms/kg/min for 30 min) were given. The haemodynamic and metabolic changes were similar in diabetics and non-diabetics. Prenalterol has the most pronounced haemodynamic effect on heart rate (increase 25-47%) and the heart rate-blood pressure-product (increase 54-81%). The metabolic effects were moderate. The slight lipolytic effect of prenalterol documented its functional selectivity for beta 1-adrenoceptors.

Echocardiographic comparison between prenalterol and dobutamine.

Prenalterol, a new cardioselective beta 1-adrenoceptor agonist, was compared with the beta-stimulator dobutamine by computer-assisted echocardiography. Prenalterol decreased preload, increased heart rate and induced an increase in contractility for a longer time in comparison with dobutamine. The small influence on blood pressure and afterload indicates selective beta 1-receptor-effect by a result of slight changes in the peripheral vascular resistance.