Analytical and preparative enantioseparations in supercritical fluid chromatography using different brands of immobilized cellulose tris (3,5-dichlorophenylcarbamate) columns: Some differences.

The aim of this study was to determine the impact of the origin and the manufacturing processes of the chiral stationary phases (CSPs) on their chromatographic behaviors. Hence, four chiral stationary phases based on immobilized tris (3,5-dichlorophenylcarbamate) derivative of cellulose supplied by four different manufacturers were evaluated. A set of twenty-nine compounds, including commercially available and in-house synthesized compounds, with a broad range of lipophilicity and polarity was chosen. Three main parameters were evaluated on all stationary phases: retention factor, selectivity and loading capacity. This work highlighted that the retention factor strongly varied according to the manufacturer. Regardless of the characteristic of the tested compounds i.e. neutral, acidic or basic, there was a trend in retention ability of the four chiral stationary phases: retention was increasing from CHIRAL ART Cellulose-SC, REFLECT I-Cellulose C, Chiralpak IC to Lux i-Cellulose-5. On the contrary, selectivity did not follow the same trend as retention. The difference in selectivity between each column towards one compound was quite low while the difference in resolution depended on the nature of the compounds investigated and was significant in certain cases. Finally, the four different columns presented similar and high loading capacity.

Ultra high performance liquid chromatography method development for separation of omeprazole and related substances on core-shell columns using a Quality by Design approach.

An updated and improved method for analysis of omeprazole/esomeprazole and related substances on core-shell columns was developed using Fusion LC Method Development™. The method was optimized with respect to column type, column temperature, mobile phase pH level, and gradient time. Four different core-shell columns were examined to develop a method suitable for both high performance- and ultra-high performance liquid chromatography using a Quality by Design approach. The final method offers two alternative columns: Poroshell EC C18 (3.0 × 100 mm, 2.7 µm) or Poroshell HPH (3.0 × 100 mm, 2.7 µm) with the same gradient elution condition and mobile phase composition. Total run time is 18 min with 12 min of gradient elution. Phosphate buffer (15 mM, pH 7.8) is selected as the aqueous mobile phase and acetonitrile as the organic mobile phase. Column temperature is set at 40°C and ultraviolet detection at 302 nm. Furthermore, by studying parameters in a systematic way, an understanding of the effect of the input parameters enhances the method robustness and should allow for regulatory flexibility in terms of post-approval changes. Compared to the current United States Pharmacopeia method, the updated method is faster, more efficient and performs well above acceptance criteria.

New Marine Antifouling Compounds from the Red Alga Laurencia sp.

Six new compounds, omaezol, intricatriol, hachijojimallenes A and B, debromoaplysinal, and 11,12-dihydro-3-hydroxyretinol have been isolated from four collections of Laurencia sp. These structures were determined by MS and NMR analyses. Their antifouling activities were evaluated together with eight previously known compounds isolated from the same samples. In particular, omaezol and hachijojimallene A showed potent activities (EC50 = 0.15-0.23 µg/mL) against larvae of the barnacle Amphibalanus amphitrite.

Novel MWCNTs/graphene oxide/pyrogallol composite with enhanced sensitivity for biosensing applications.

A novel and highly sensitive biosensor employing graphene oxide nano-sheets (GO), multiwalled carbon nanotubes (MWCNTs), and pyrogallol (PG) was fabricated and utilized for the sensitive determination of omeprazole (OME). The morphological and structural features of the composite material were characterized using different techniques. The modified electrode showed a remarkable improvement in the anodic oxidation activity of OME due to the enhancement in the current response compared to the bare carbon paste electrode (CPE). Sensor composition and measurement conditions were optimized using an experimental design. Differential pulse voltammetry (DPVs) exhibited two expanded linear dynamic ranges of 2.0×10-10-6.0×10-6M and 6.0×10-6-1.0×10-4M for OME at pH 7 with a detection limit of 1.02×10-11M. The practical analytical utilities of the modified electrode were demonstrated by the accurate determination of OME in pharmaceutical formulation and human serum samples with mean recoveries of 100.97% and 98.58%, respectively. The results clearly revealed that the proposed sensor is applicable to clinical analysis, quality control and routine determination of drugs in pharmaceutical formulations.

A practical approach for predicting retention time shifts due to pressure and temperature gradients in ultra-high-pressure liquid chromatography.

Large pressure gradients are generated in ultra-high-pressure liquid chromatography (UHPLC) using sub-2µm particles causing significant temperature gradients over the column due to viscous heating. These pressure and temperature gradients affect retention and ultimately result in important selectivity shifts. In this study, we developed an approach for predicting the retention time shifts due to these gradients. The approach is presented as a step-by-step procedure and it is based on empirical linear relationships describing how retention varies as a function of temperature and pressure and how the average column temperature increases with the flow rate. It requires only four experiments on standard equipment, is based on straightforward calculations, and is therefore easy to use in method development. The approach was rigorously validated against experimental data obtained with a quality control method for the active pharmaceutical ingredient omeprazole. The accuracy of retention time predictions was very good with relative errors always less than 1% and in many cases around 0.5% (n=32). Selectivity shifts observed between omeprazole and the related impurities when changing the flow rate could also be accurately predicted resulting in good estimates of the resolution between critical peak pairs. The approximations which the presented approach are based on were all justified. The retention factor as a function of pressure and temperature was studied in an experimental design while the temperature distribution in the column was obtained by solving the fundamental heat and mass balance equations for the different experimental conditions. We strongly believe that this approach is sufficiently accurate and experimentally feasible for this separation to be a valuable tool when developing a UHPLC method. After further validation with other separation systems, it could become a useful approach in UHPLC method development, especially in the pharmaceutical industry where demands are high for robustness and regulatory oversight.

Electrochemical incineration of omeprazole in neutral aqueous medium using a platinum or boron-doped diamond anode: degradation kinetics and oxidation products.

The electrochemical incineration of omeprazole, a widely prescribed gastrointestinal drug which is detected in natural waters, has been studied in a phosphate buffer of pH 7.0 by anodic oxidation with electrogenerated H(2)O(2) (AO-H(2)O(2)) operating at constant current density (j). The experiments were carried out in a cell equipped with either a Pt or a boron-doped diamond (BDD) anode and an air-diffusion cathode to continuously produce H(2)O(2). In these systems, organics are mainly oxidized by hydroxyl radicals formed at the Pt or BDD surface from water oxidation. A partial total organic carbon (TOC) abatement close to 78% for omeprazole was achieved by AO-H(2)O(2) with a BDD anode after consumption of 18 Ah L(-1) at 100 mA cm(-2), whereas the alternative use of Pt did not allow mineralizing the drug. However, the drug was totally removed using both anodes, although it decayed more rapidly using BDD. In this latter system, increasing j accelerated the degradation process, but lowering the mineralization current efficiency. Greater drug content also enhanced the degradation rate with higher mineralization degree and current efficiency. The kinetics for omeprazole decay always followed a pseudo-first-order reaction and its rate constant increased with increasing j and with decreasing its concentration. Seven heteroaromatic intermediates and four hydroxylated derivatives were detected by LC-MS, while nine short-linear carboxylic acids were identified and quantified by ion-exclusion HPLC. These acids were largely accumulated using Pt and rapidly removed using BDD, thus explaining the partial mineralization of omeprazole achieved by AO-H(2)O(2) with the latter anode. The release of inorganic ions such as NO(3)(-), NH(4)(+) and SO(4)(2-) was followed by ionic chromatography. A plausible reaction sequence for omeprazole mineralization involving all intermediates detected is proposed.

RP-HPLC/pre-column derivatization for analysis of omeprazole, tinidazole, doxycycline and clarithromycin.

A validated, reliable and accurate reversed-phase high performance liquid chromatographic method using pre-column derivatization was adopted for the simultaneous determination of two ternary mixtures containing omeprazole, tinidazole and doxycycline hyclate or clarithromycin. Separation was achieved on a C18 column, through a gradient elution system using acetonitrile-methanol-water adjusted to pH = 6.60. Drugs were detected at 277 nm over concentration ranges of 1-112, 5-125, 2.5-550 and 2.5-100 µg/mL for omeprazole, tinidazole, doxycycline hyclate and clarithromycin, respectively. This is the first method that has isolated and identified clarithromycin derivative by infrared and mass spectroscopy. This method is the first study for the simultaneous determination of omeprazole, tinidazole, doxycycline hyclate and clarithromycin in combined mixtures and pharmaceutical formulations.

Removal of trace organic chemical contaminants by a membrane bioreactor.

Emerging wastewater treatment processes such as membrane bioreactors (MBRs) have attracted a significant amount of interest internationally due to their ability to produce high quality effluent suitable for water recycling. It is therefore important that their efficiency in removing hazardous trace organic contaminants be assessed. Accordingly, this study investigated the removal of trace organic chemical contaminants through a full-scale, package MBR in New South Wales, Australia. This study was unique in the context of MBR research because it characterised the removal of 48 trace organic chemical contaminants, which included steroidal hormones, xenoestrogens, pesticides, caffeine, pharmaceuticals and personal care products (PPCPs). Results showed that the removal of most trace organic chemical contaminants through the MBR was high (above 90%). However, amitriptyline, carbamazepine, diazepam, diclofenac, fluoxetine, gemfibrozil, omeprazole, sulphamethoxazole and trimethoprim were only partially removed through the MBR with the removal efficiencies of 24-68%. These are potential indicators for assessing MBR performance as these chemicals are usually sensitive to changes in the treatment systems. The trace organic chemical contaminants detected in the MBR permeate were 1 to 6 orders of magnitude lower than guideline values reported in the Australian Guidelines for Water Recycling. The outcomes of this study enhanced our understanding of the levels and removal of trace organic contaminants by MBRs.

Enantioseparation of omeprazole--effect of different packing particle size on productivity.

Enantiomeric separation of omeprazole has been extensively studied regarding both product analysis and preparation using several different chiral stationary phases. In this study, the preparative chiral separation of omeprazole is optimized for productivity using three different columns packed with amylose tris (3,5-dimethyl phenyl carbamate) coated macroporous silica (5, 10 and 25 µm) with a maximum allowed pressure drop ranging from 50 to 400 bar. This pressure range both covers low pressure process systems (50-100 bar) and investigates the potential for allowing higher pressure limits in preparative applications in a future. The process optimization clearly show that the larger 25 µm packing material show higher productivity at low pressure drops whereas with increasing pressure drops the smaller packing materials have substantially higher productivity. Interestingly, at all pressure drops, the smaller packing material result in lower solvent consumption (L solvent/kg product); the higher the accepted pressure drop, the larger the gain in reduced solvent consumption. The experimental adsorption isotherms were not identical for the different packing material sizes; therefore all calculations were recalculated and reevaluated assuming identical adsorption isotherms (with the 10 µm isotherm as reference) which confirmed the trends regarding productivity and solvent consumption.

Enantioseparation of rabeprazole and omeprazole by nonaqueous capillary electrophoresis with an ephedrine-based ionic liquid as the chiral selector.

An ephedrine-based chiral ionic liquid, (+)-N,N-dimethylephedrinium-bis(trifluoromethanesulfon)imidate ([DMP](+) [Tf(2) N](-) ), served as both chiral selector and background electrolyte in nonaqueous capillary electrophoresis. The enantioseparation of rabeprazole and omeprazole was achieved in acetonitrile-methanol (60:40 v/v) containing 60 mm[DMP](+) [Tf(2) N](-) . The influences of separation conditions, including the concentration of [DMP](+) [Tf(2) N](-) , the electrophoretic media and the buffer, on enantioseparation were evaluated. The mechanism of enantioseparation was investigated and discussed. Ion-pair interaction and hydrogen bonding may be responsible for the main separation mechanism.

Direct HPLC enantioseparation of omeprazole and its chiral impurities: application to the determination of enantiomeric purity of esomeprazole magnesium trihydrate.

Analytical and semipreparative high-performance liquid chromatography (HPLC) enantioseparation of the proton-pump inhibitor omeprazole (OME) and its potential organic chiral impurities were accomplished on the immobilised-type Chiralpak IA chiral stationary phase (CSP) under both polar organic and normal-phase conditions. The (S)-enantiomers were isolated with a purity of >99% ee and their absolute configuration was empirically assigned by circular dichroism (CD) spectroscopy. A chemo- and enantioselective HPLC method was validated to control the enantiomeric purity of the (S)-enantiomer of OME (ESO), an active ingredient contained in drug products, in the presence of chiral and achiral related substances. The precision, linearity and accuracy of the determination of the (R)-impurity as well as the recovery of ESO from a pharmaceutical preparation were determined. The proposed method uses the mixture methyl tert-butylether (MtBE)-ethyl acetate (EA)-ethanol (EtOH)-diethylamine (DEA) 60:40:5:0.1 (v/v/v/v) as a mobile phase. In these conditions, linearity over the concentration range 0.5-25 microg/ml for (R)-enantiomer was obtained. The limits of detection and quantification were 99 and 333 ng/ml, respectively. The intra and inter-day assay precision was less than 2% (RSD%).

Use of (S)-BINOL as NMR chiral solvating agent for the enantiodiscrimination of omeprazole and its analogs.

The application of (S)-1,1'-binaphthyl-2,2'-diol as NMR chiral solvating agent (CSA) for omeprazole, and three of its analogs (lanso-, panto-, and rabe-prazole) was investigated. The formation of diastereomeric host-guest complexes in solution between the CSA and the racemic substrates produced sufficient NMR signal splitting for the determination of enantiomeric excesses by (1)H- or (19)F-NMR spectroscopy. Using of hydrophobic deuterated solvents was mandatory for obtaining good enantiodiscrimination, thus suggesting the importance of intermolecular hydrogen bonds in the stabilization of the complexes. The method was applied to the fast quantification of the enantiomeric purity of in-process samples of S-omeprazole.

Enantioseparation of omeprazole and its metabolite 5-hydroxyomeprazole using open tubular capillary electrochromatography with immobilized avidin as chiral selector.

The present paper demonstrates the enantiomeric separation of omeprazole and its metabolite 5-hydroxyomeprazole performed with open tubular capillary electrochromatography (OT-CEC). The protein avidin was used as the chiral selector. Avidin was immobilized by a Schiffs base type of reaction where the protein was via glutaraldehyde covalently bonded to the amino-modified wall of a fused-silica capillary, 50 microm i.d. Both racemates were baseline resolved. Resolution was 1.9 and 2.3, respectively, using ammonium acetate buffer, pH 5.8, 5% methanol, with UV-detection. These values of resolution using OT-CEC are higher than earlier published results regarding chiral separation of omeprazole and 5-hydroxyomeprazole on packed CEC. The number of theoretical plates also indicated good separation efficiency.

[Chiral separation of tenatoprazole enantiomers using high performance liquid chromatography on vacomycin-bonded chiral stationary phase].

Vacomycin-bonded chiral stationary phase was used for the direct chiral separation of tenatoprazole enantiomers using reversed-phase high performance liquid chromatography (HPLC). The influences of the kinds and concentration of buffer and organic modifier, the pH value of buffer, column length and column temperature on the separation were examined. The chiral HPLC method for the separation of tenatoprazole enantiomers on a Chirobiotic V column (150 mm x 4.6 mm, 5 microm) was established with simplicity and good reproducibility using 0.02 mol/L ammonium acetate buffer (pH 6.0)-tetrahydrofuran (93:7, v/v) as the mobile phase at a flow rate of 0.5 mL/min and 20 degrees C. Under the above conditions, the enantiomers were separated on baseline with the resolution of 1.68. The relative standard deviations (RSDs) for the retention times of tenatoprazole enantiomers were 0.48% and 0.49% (n = 6). The RSDs for the peak areas of tenatoprazole enantiomers were 0.45% and 0.55% (n = 6).

Enantioselective quantification of omeprazole and its main metabolites in human serum by chiral HPLC-atmospheric pressure photoionization tandem mass spectrometry.

Omeprazole is a proton pump inhibitor drug in widespread use for the reduction of gastric acid production. It is also proposed as a test substance for the phenotyping of cytochrome CYP3A4 and CYP2C19 enzyme activities. For this purpose, it is necessary to quantify, additionally to omeprazole, the two main metabolites 5-hydroxyomeprazole and omeprazole-sulfon in human plasma. Since omeprazole is a racemic mixture of two enantiomers and its enzymatic decomposition depends in part on its chiral configuration, full information about its metabolic breakdown can only be gained by enantioselective quantification of the drug and its metabolites. We introduce a new LC-MS/MS method that is capable to simultaneously quantify omeprazole and its two main metabolites enantioselectively in human serum. The method features solid-phase extraction, normal phase chiral HPLC separation and atmospheric pressure photoionization tandem mass spectrometry. As internal standards serve stable isotope labeled omeprazole and 5-hydroxyomeprazole. The calibration functions are linear in the range of 5-750 ng/ml for the omeprazole enantiomers and omeprazole-sulfon, and 2.5-375 ng/ml for the 5-hydroxyomeprazole enantiomers, respectively. Intra- and inter-day relative standard deviations are <7% for omeprazole and 5-hydroxyomeprazole enantiomers, and <9% for omeprazole-sulfon, respectively.

High-performance liquid chromatographic separation of rolipram, bupivacaine and omeprazole using a tartardiamide-based stationary phase influence of flow rate and temperature on the enantioseparation.

Chromatographic separation of the chiral drugs rolipram, bupivacaine and omeprazole on a tartardiamide-based stationary phase commercially named Kromasil CHI-TBB is shown in this work. The effect of temperature on the chromatographic separation of the chiral drugs using the Kromasil CHI-TBB stationary phase was determined quantitatively so as to contribute toward the design for the racemic mixtures of the named compound by using chiral columns. A decrease in the retention and selectivity factors was observed, when the column temperature increased. Van't Hoff plots provided the thermodynamic data. The variation of the thermodynamic parameters enthalpy and entropy are clearly negative meaning that the separation is enthalpy controlled.

Semipreparative enantiomeric separation of omeprazole by supercritical fluid chromatography.

Omeprazole, a widely used antiulcer drug, has been enantiomerically separated at semipreparative scale on a polysaccharide based chiral stationary phase by supercritical fluid chromatography (SFC). For this work, a modular supercritical fluid chromatograph was adapted to operate at semipreparative scale and a Chiralpak AD (250 mm x 10 mm) column was used. The effect of two organic modifiers (ethanol and 2-propanol) was studied, and different injection volumes and concentrations of the omeprazole racemic mixture were evaluated in order to obtain high enantiomeric purities and production rates. Better results were achieved using concentration overloading instead of volume overloading. The recoveries decreased when the requirements of enantiomeric purity or the load increased, but it was possible to recover 100% of both enantiomers at an enantiomeric purity higher than 99.9% under some loading conditions, like injecting 1 and 2 ml of a solution of 3g/l. As far as production rates are concerned, the best result for S-(-)-omeprazole at that purity (27.2mg/h) was achieved with sample concentrations of 10 g/l and the injection of 2 ml, while a volume of 4 ml was better in the case of R-(+)-omeprazole (20.5mg/h).

Determination of lansoprazole and two of its metabolites by liquid-liquid extraction and automated column-switching high-performance liquid chromatography: application to measuring CYP2C19 activity.

A simple and sensitive column-switching high-performance liquid chromatographic (HPLC) method for the simultaneous determination of lansoprazole, a proton pump inhibitor and its major metabolites: 5-hydroxylansoprazole and lansoprazole sulfone in human plasma. The test compounds were extracted from 1 mL of plasma using diethyl ether-dichloromethane (7:3, v/v) mixture and the extract was injected into a column I (TSK-PW precolumn, 10 microm, 3.5 mm x 4.6 mm i.d.) for clean-up and column I (C(18) STR ODS-II analytical column, 5 microm, 150 mm x 4.6 mm i.d.) for separation. The peak was detected by a ultraviolet detector set at a wavelength of 285 nm, and the total time for a chromatographic separation was approximately 25 min. The method was validated for the concentration range from 3 to 5000 ng/mL. Mean recoveries were 74.0% for lansoprazole, 68.3% for 5-hydroxylansoprazole, and 79.4% for lansoprazole sulfone. Intra- and inter-day relative standard derivatives were less than 6.1 and 5.1% for lansoprazole, 5.8 and 5.8% for 5-hydroxylansoprazole, 4.4 and 5.9% for lansoprazole sulfone, respectively, at the different concentration ranges. This method is suitable for use in therapeutic drug monitoring and pharmacokinetic studies, and provides use tool for measuring CYP2C19 activity.

Pharmacokinetic differences between lansoprazole enantiomers in rats.

Because limited information is available about potential differences between the pharmacokinetics and pharmacodynamics of the enantiomers of lansoprazole, the enantioselective pharmacokinetics of the compound have been investigated in rats. There was a noticeable difference between the serum levels of the enantiomers of lansoprazole and of their metabolites, 5-hydroxylansoprazole enantiomers, after oral administration of the racemate (50 mg kg(-1)) to rats. Cmax (maximum serum concentration) and AUC (area under the serum concentration-time curve) for (+)-lansoprazole were 5-6 times greater than those for (-)-lansoprazole, whereas for (+)-5-hydroxylansoprazole both values were significantly smaller than those for the (-) enantiomer. CLtot/F values (where CLtot is total clearance and F is the fraction of the dose absorbed) for (+)-lansoprazole were significantly smaller than those for the (-) enantiomer. There was no significant difference between the absorption rate constants of the lansoprazole enantiomers in the in-situ absorption study. The in-vitro protein-binding study showed that binding of (+)-lansoprazole to rat serum proteins was significantly greater than for the (-) enantiomer. The in-vitro metabolic study showed that the mean metabolic ratio (45.9%) for (-)-lansoprazole was significantly greater than that (19.8%) for the (+) enantiomer in rat liver microsomes at 5.6 microM lansoprazole. These results show that the enantioselective disposition of lansoprazole could be a consequence of the enantioselectivity of plasma-protein binding and the hepatic metabolism of the enantiomers.

Chiral resolution of pantoprazole sodium and related sulfoxides by complex formation with bovine serum albumin in capillary electrophoresis.

The separation of enantiomers of pantoprazole sodium, omeprazole and lansoprazole by capillary zone electrophoresis using bovine serum albumin (BSA) as the chiral selector is described. Baseline separation of the three structurally related drugs was obtained after optimization of the most important experimental parameters. For this purpose, influences such as BSA concentration, pH and concentration of 1-propanol as organic modifier on the separation were investigated. Increasing concentrations of BSA improved the chiral resolution but lowered the sensitivity of the detection system. Discrimination of the enantiomers was observed only in a narrow pH range of 7-8. An optimum of pH 7.4 was a good compromise in terms of enantio-resolution and peak shape. 1-Propanol when added to the buffer system, improved the peak shape of the analytes and the resolution. The optimized method has been validated for pantoprazole sodium and is useful for routine analysis.