Per-Olof Lagerström

Determination of omeprazole and metabolites in plasma and urine by liquid chromatography

Omeprazole, a substituted benzimidazole and a new gastric acid inhibitor, has been determined in plasma and urine, together with three of its metabolites--the sulphide, the sulphone and the hydroxy compound. The methods comprise extraction from the biological materials with methylene chloride, followed either by direct injection of the extract onto a normal-phase liquid chromatography column or evaporation, dissolution and injection onto a reversed-phase system. The compounds were detected using ultraviolet spectrometry. The absolute recoveries obtained were mostly above 95%. The minimum determinable concentration for omeprazole was 20 nmol/l in plasma (relative standard deviation 10-15%) and 50 nmol/l in urine. The metabolites could also be determined at the same levels.

Reversed retention order and other stereoselective effects in the separation of amino alcohols on Chiralcel OD

Abstract The enantioselective resolution of a series of amino alcohols on Chiralcel OD was studied with respect to the effect of temperature, alcohol additive and water content in a mobile phase of hexane with added diethylamine. The chain length between the hydroxy and the amino groups in the solutes had a considerable influence on the stereoselectivity. For one of the amino alcohols a reversal of the retention order between the antipodes was obtained by varying the above parameters. The amino alcohols are retained by at least two chiral sites, one of which is highly dependent on hydrogen bonding. This bonding ability can be directly controlled by the water content of the mobile phase.

Stereoselective effects in the separation of enantiomers of omeprazole and other substituted benzimidazoles on different chiral stationary phases

Abstract The enantioselective separation of omeprazole on different chiral stationary phases was investigated. The two enantiomers could be resolved on three different phases with immobilized protein, Chiral-AGP, Ultron ES-OVM and BSA-DSC, employing aqueous mobile phases with 2-propanol as organic modifier. On Chiralpak AD, an amylose-based chiral stationary phase, the enantiomers of omeprazole and three analogues could be separated using a non-polar hexane-ethanol mobile phase. For omeprazole the retention order was reversed when 2-propanol was replaced with ethanol or methanol as the modifier of hexane in the mobile phase.

The pharmacokinetics of omeprazole in humans--a study of single intravenous and oral doses.

The pharamacokinetics of omeprazole, hydroxyomeprazole, omeprazolesulfone, and “remaining metabolites” have been studied in eight young healthy subjects following an acute i.v. and oral dose of 10 and 20 mg of 14C-labeled drug, respectively. The oral dose was given as a buffered solution. Two subjects exhibited essentially higher and more sustained plasma levels of omeprazole than the others. This was due to a higher bioavailability, lower clearance, and longer t1/2 of omeprazole in these two subjects. Maximum concentration (0.7–4.6 mUmol/L) was reached between 10 and 25 min after oral dosing. The median bioavailability was 39% (25–117%) and the median systemic plasma clearance was 624 ml/min (range of 59–828 ml/min). The corresponding t1/2 for the i.v. dose was 35 min (16–150 min) and 39 min (14–186 min) after oral administration. The drug was rapidly distributed to extravascular sites (mean t1/2Λ1 = 3.0 \pm 0.8 min). Mean Vss was 0.23 \pm 0.04 L/kg. Low systemic clearance of omeprazole was associated with a decreased formation rate of hydroxyomepraxole and “remaining metabolites” while omeprazole-sulfone formation seemed to be less affected. However, there was a clear-cut correlation between the t1/2 of omeprazole and of its omeprazolesulfone metabolite, indicating that the elimination of these two compounds is mediated by the same isoenzyme. The mean urinary recovery of the radioactive dose during 96 h was 78.3 \pm 2.3 and 75.7 \pm 2.6% for the i.v. and oral dose, respectively. Insignificant amounts were due to unchanged drug and omeprazolesulfone. The excretion of hydroxyomeprazole during the first 12 h varied between 4.6 to 15.5% of a given dose. The mean recovery of radioactivity in the feces was 19.3 \pm 3.1% of a given i.v. dose and 18.2 \pm 2.3% when given orally. It is concluded that omeprazole is mainly eliminated metabolically and that there is a substantial interindividual variation in the rate of formation of primary and secondary metabolites. This variation in omeprazole disposition is probably of limited clinical importance. The half-life, with a maximum of 3 h, is too short to cause accumulation when the drug is administered in a once-daily regimen.

A study of the interaction between omeprazole and phenytoin in epileptic patients.

This study was performed to determine the effect of omeprazole, given in therapeutically recommended doses, on the steady-state plasma levels of phenytoin in epileptic patients. Five men and three women of median age 34 years participated in the study. Steady-state plasma levels of phenytoin were measured once a week for 2 weeks before and after, respectively, and during 3 weeks of concomitant omeprazole treatment with 20 mg daily. Urinary excretion of phenytoin and its metabolite [5-(p-hydroxyphenyl)-5-phenyl-hydantoin were determined before and at the end of the omeprazole treatment period. The steady-state plasma phenytoin levels as well as urinary excretion of phenytoin and its main metabolite were unchanged during omeprazole treatment. The results from this study suggest that concomitant omeprazole treatment in therapeutically recommended doses (20 mg daily) will not significantly affect the steady-state plasma levels of phenytoin in epileptic patients.

Omeprazole Treatment Does Not Affect the Metabolism of Caffeine

This study was performed to investigate the possible influence of repeated omeprazole dosing on the metabolism of caffeine, which has been shown to reflect the activity of one specific enzyme within the hepatic cytochrome P450 family, P450IA2. Ten healthy, nonsmoking young men participated in this placebo-controlled double-blind trial. Each subject was given omeprazole, 20 mg, every morning for 1 week and placebo every morning for 1 week in random order and separated by a 2-3 week washout period. On the sixth and seventh days of each period urine was collected twice daily, and urinary metabolites of caffeine were determined by high-performance liquid chromatography. The urinary metabolite ratio of three paraxanthine 7-demethylation products relative to a paraxanthine-hydroxylation product corresponds to caffeine clearance and, therefore, to P450IA2 activity. This calculated ratio was 4.8 (95% confidence interval, 3.9-5.6) in the placebo and 4.6 (95% confidence interval, 3.6-5.5) in the omeprazole period. These results show that the metabolism of caffeine was unaltered following omeprazole treatment, indicating that omeprazole treatment has no influence on cytochrome P450IA2 activity in the clinical situation.

Electron-capture—gas chromatographic determination of atenolol in plasma and urine, using a simplified procedure with improved selectivity

A sensitive gas chromatographic method for the quantitative analysis of atenolol in human plasma and urine is described. Atenolol is extracted with dichloromethane containing heptafluorobutanol to improve the extraction ability. Derivatization with trifluoroacetic anhydride in diethyl ether gives a bistrifluoroacetyl derivative which is more selectively detected by an electron-capture detector than is the corresponding heptafluorobutyryl derivative. The method allows determination down to 20 nmol/l (5 ng/ml) in 1 ml of sample with a relative standard deviation below 10%.

High-performance liquid chromatographic assay for human liver microsomal omeprazole metabolism

Assays for the measurement of omeprazole metabolites in plasma and urine have been reported, but when applied to the determination of omeprazole metabolites formed by human liver microsomal incubations there were obvious limitations in sensitivity. The present high-performance liquid chromatographic (HPLC) assay, which comprises extraction, evaporation and reconstitution, is several-fold more sensitive with a limit of detection of approximately 2 pmol (2 nM in incubate) for omeprazole sulphone and 25 pmol (25 nM in incubate) for hydroxyomeprazole. Extraction efficiency is essentially quantitative and is highly reproducible (coefficient of variation = 2.1% for both metabolites). The assay is linear over a wide range of concentrations and the formation of the metabolites is linear with respect to both time (to 15 min) and protein concentration (to 1.5 mg/ml). Two minor metabolites, one of which was identified tentatively as 5-O-desmethylomeprazole, were also formed by human liver microsomes and could be determined by this method. Preliminary studies of the formation of omeprazole sulphone and hydroxyomeprazole showed that the formation kinetics in human liver microsomes were biphasic for both metabolites, suggesting that at least two different cytochrome P450 isoforms are involved in their formation.

Drug level monitoring: cardiovascular drugs

Methods for the determination of cardiovascular drugs in blood and plasma are critically reviewed with emphasis on gas and liquid chromatographic techniques. The importance of the various procedures is discussed, in particular sample work-up where the conditions for isolation and derivatization of the compounds are decisive for the accuracy and precision of the methods. Compared with other assay techniques chromatographic methods are generally to be preferred owing to their better selectivity. In the review the following groups are discussed: digitalis glycosides, antiarrhythmic agents, beta-adrenoceptor antagonists, vasodilating agents, antihypertensive compounds, and diuretics.

Determination of metaprolol and two major metabolites in plasma and urine by column liquid chromatography and fluorometric detection

Metoprolol and its alpha-hydroxy metabolite were determined in plasma down to 2 nmol/l (S.D. 10-15%) after solvent extraction and bonded-phase liquid chromatography with fluorometric detection. The major metabolite with a carboxylic function was also measured in plasma when liquid-solid extraction on a column activated with dodecyl sulphate was applied. In urine the three components were assayed by direct injection of a diluted sample.