Determination of remoxipride in human plasma and urine by reversed-phase ion-pair high-performance liquid chromatography.

A sensitive method is described for the measurement of remoxipride in human plasma and urine. Remoxipride and its internal standard are extracted from plasma or urine at pH 12 with a mixture of hexane and methyl tert.-butyl ether. After washing the organic phase with base, the compounds are extracted into acid and analyzed on a C18 column with ultraviolet detection at 214 nm. The mobile phase is composed of acetonitrile and aqueous buffer (sodium perchlorate and phosphoric acid, pH 1.7). The limits of reliable quantitation for remoxipride are 12.5 and 50 ng/ml for plasma and urine, respectively. The run times are 6 min for plasma and 3 min for urine. The method has been successfully used to assay remoxipride clinical study samples. This mobile phase has also been successfully applied to the analysis of other basic drugs such as cimetidine, codeine, diltiazem and quinidine with minor modifications.     

Determination of ranitidine and its metabolites in human urine by reversed-phase ion-pair high-performance liquid chromatography

A method using ion-pair high-performance liquid chromatography is presented for determining ranitidine, ranitidine N-oxide, ranitidine S-oxide and desmethyl ranitidine in the urine from four volunteers, given on separate occasions an intravenous and oral dose of 100 mg ranitidine. This method has been used to study the metabolism and pharmacokinetics of ranitidine by man. It was found that the elimination half-life of ranitidine ranged from 110-246 min. The mean renal clearance of ranitidine in these four volunteers was 512 ml/min.     

Determination of pemoline in plasma, urine and tissues by reversed-phase high-performance liquid chromatography

A simple and selective high-performance liquid chromatographic method with ultraviolet detection at 215 nm for the determination for pemoline in rat plasma, urine and tissues is described. Pemoline in the samples was extracted with methylene chloride at pH 10 and the organic phase was evaporated after adding 5-methyl-5-phenylhydantoin used as an internal standard. Pemoline and the internal standard were separated on a Kaseisorb LC C8-60-5 reversed-phase column. The limits of determination of pemoline in 0.1-0.2 ml of plasma, urine and tissue homogenates were 2, 100 and 20 ng, respectively. The method should be useful for studies of the pharmacokinetics and distribution of pemoline in small animals.     

Determination of teicoplanin in human plasma and urine by affinity and reversed-phase high-performance liquid chromatography

A sensitive, highly selective and simple high-performance liquid chromatographic method for the determination of teicoplanin, a novel glycopeptide antibiotic, composed of six components, in human plasma and urine is described. After an isolation step by affinity chromatography, the antibiotic substances were chromatographed on a Nucleosil C18 column with phosphate buffer-acetonitrile according to a gradient profile. All the components were detected by their UV absorption at 240 nm. The concentration of teicoplanin was determined by using the external standard procedure. This method was applied to the sum of the six major components as well as to each of them separately. The linearity of the method was checked between 0.5 and 50 micrograms/ml for plasma and between 2 and 50 micrograms/ml for urine. The limit of detection was 0.1 microgram/ml for both biological fluids. The coefficients of variation of the between-day assays did not exceed 8.6 and 8.9% in plasma and urine, respectively. The application of the method to a pharmacokinetic study of teicoplanin after a single intravenous therapeutic dose in a patient is reported. This rapid technique also appears to be suitable for drug monitoring.     

Determination of nitrophenol derivatives in various crops by reversed-phase ion-pair high-performance liquid chromatography

A method for the determination of nitrophenol derivatives in various crops and soil is described. The sample is extracted with dichloromethane; the extract is evaporated to dryness and the residue is dissolved in an alkaline methanol/water mixture. After filtration this solution is injected. The eluent contains methanol, water, a phosphate buffer and hexadecyltrimethylammonium as the pairing ion. Blank chromatograms for various crops do not show interfering peaks with detection at 365 or 405 nm. The limit of detection is usually around 0.01 mg kg^-1; recoveries generally exceed 80% (coefficient of variation, 5–10%). For 2,4-dinitrophenylthiocyanate and dinocap, recoveries are somewhat lower.     

Determination of L-alanosine in plasma and urine by reversed-phase high-performance liquid chromatography of the Dns derivative

L-Alanosine is an antitumour antibiotic that has recently been placed in clinical trial. We have developed a relatively rapid and specific assay for urinary and plasma alanosine, based on formation of the Dns derivative and separation of this from other Dns compounds by reversed-phase high-performance liquid chromatography. Dns-Alanosine is detected by its absorption at 254 nm, since alanosine is atypical in that it forms a Dns derivative with very low fluorescence. The lower limit of detection of alanosine in plasma is 0.1 microgram/ml. The assay has been used to measure the levels of alanosine in the plasma and urine of rabbits and of man.     

Determination of carnosine and other biogenic imidazoles in equine plasma by isocratic reversed-phase ion-pair high-performance liquid chromatography.

The isocratic reversed-phase ion-pair high-performance liquid chromatographic technique presented provides a sensitive, rapid and reproducible analytical method for the selective determination of carnosine and other biogenic imidazoles in equine plasma. Plasma was deproteinized with 5-sulphosalicylic acid and the compounds of interest were isolated by sorbent extraction on Bond Elut PRS cartridges. Recoveries were 97-105% and the lowest limits of detection were 58.3-80.1 nM. All compounds of interest were well resolved within a maximum retention time of 9.2 min. The mean equine plasma carnosine level determined by this method was 11.31 microM. Comparative determinations were made in canine and human plasma. Carnosine was not detected in human plasma. Concentrations of imidazole in canine plasma are reported here for the first time.     

Determination of paracetamol and its four major metabolites in mouse plasma by reversed-phase ion-pair high-performance liquid chromatography.

A reversed-phase ion-pair high-performance liquid chromatographic method has been used for the separation of paracetamol and its four major metabolites (glucuronide, sulphate, cysteine and mercapturate conjugates) in mouse plasma samples. An ODS column was used and the mobile phase consisted of an aqueous solution of 0.01 M tetrabutylammonium chloride and 0.01 M Tris buffered to pH 5.0 with phosphoric acid, with methanol as the organic solvent. The gradient elution started with 30% methanol. After a delay of 0.5 min the methanol concentration was increased linearly to 75% over 7.5 min. The column was returned to the initial conditions after a delay of 1 min. A methanol solution of theophylline was added to the mouse plasma sample, centrifuged and immediately injected into the chromatographic system. The advantages of this method include good and rapid separation (last metabolite detected at 6.86 min), well resolved peaks, only a small amount of sample required for assay, adequate precision (no coefficient of variation was greater than 10% for paracetamol metabolites) and a high sensitivity (particularly for unchanged paracetamol and the cysteine conjugate).     

Identification of cosmetic dyes by ion-pair reversed-phase high-performance liquid chromatography

A method based on ion-pair reversed-phase high-performance liquid chromatography with detection at four wavelengths between 400 and 600 nm is reported for the separation and identification of the most common synthetic colour additives in cosmetic products. All the dyes generally employed in the U.S.A. and almost all those in current use in cosmetics in the European Community have been taken into account. The chromatography was performed on a C8 bonded silica packed column, with a 60-min gradient changing from 10 to 95% acetonitrile in water containing 10(-2) M sodium perchlorate (pH 3.0) as mobile phase (flow-rate 2.5 ml/min). Detection limits are in the range 20-100 ng for all dyes investigated. The method has been applied to the analysis of commercial lipsticks.     

Determination of cortisol in human plasma by reversed-phase high-performance liquid chromatography

A method is described for the measurement of cortisol in human plasma using 45% aqueous methanol eluent on a 120 mm x 4.5 mm I.D. Hypersil octadecylsilane column with UV detection at 239 nm after a simple dichloromethane extraction and evaporation with a prednisone internal standard. The sample preparation time and chromatography time are each about 15 min and linear correlations have been obtained with plasma samples assayed by the Mattingly fluorimetric technique and a commercial-kit competitive protein binding method. Concentration down to 30 nmol/l may be measured and the method can be used when fluorimetry is invalidated by interference, particularly from spironolactone.     

Determination of p-chloronitrobenzene in plasma by reversed-phase high-performance liquid chromatography.

A simple, accurate and precise isocratic reversed-phase high-performance liquid chromatographic method was developed and validated for the determination of p-chloronitrobenzene (p-CNB) in rat plasma. A plasma sample was deproteinized with methanol containing the internal standard (p-bromonitrobenzene). The resulting methanol eluate obtained after centrifugation was filtered and injected into a high-performance liquid chromatograph (50 microliters each). A column packed with 5 microns octadecylsilane (ODS) spherical particles was used with isocratic elution of methanol-water (45:55, v/v) at a flow-rate of 1.0 ml/min. The compounds were detected by ultraviolet absorbance at 280 nm. The retention times of p-CNB and the internal standard were 12.5 and 15.5 min, respectively, at a column oven temperature of 30 degrees C. The results were linear from 0.05 to 100 micrograms/ml (r = 0.999), and the detection limit was 0.01 microgram/ml. The relative error and the coefficient of variation on replicate assays were less than 7 and 10%, respectively, for all concentrations studied. The overall recoveries of p-CNB were between 97 and 105%. Plasma samples could be stored for up to one month at -20 degrees C.     

Determination of bromazepam in plasma and of its main metabolites in urine by reversed-phase high-performance liquid chromatography

A high-performance liquid chromatographic method for the determination of bromazepam in plasma and of its main metabolites in urine is described. The unchanged drug is extracted from plasma with dichloromethane, using Extrelut 1 extraction tubes. The residue from this extract is subsequently analysed by reversed-phase high-performance liquid chromatography with ultraviolet detection (230 nm). The limit of detection is 6 ng/ml of plasma, using a 1-ml specimen. For the determination of the metabolites, the urine samples are incubated to effect enzymatic deconjugation and are then extracted with dichloromethane. Following two clean-up steps (back extractions), the final residue is analysed on the same reversed-phase system as the plasma samples. The limit of detection for the two metabolites is 200 ng/ml.     

Determination of remoxipride in plasma and urine by reversed-phase column liquid chromatography

A reversed-phase column liquid chromatographic method for the determination of remoxipride, a novel antipsychotic drug, in biological fluids is described. A simple one-step extraction is used followed by liquid chromatography on a 3-microns octadecylsilica column and ultraviolet absorbance detection. The method is accurate and precise for clinical remoxipride levels in both plasma and urine. For situations where a higher sensitivity is necessary a two-step extraction and a modified mobile phase are used. With this modification plasma concentrations down to 2 nM can be determined with acceptable precision.     

Determination of amdinocillin in plasma and urine by high-performance liquid chromatography

A rapid, sensitive and specific high-performance liquid chromatographic (HPLC) assay was developed for the determination of amdinocillin (formerly mecillinam) in human plasma and urine. The assay is performed by direct injection of a plasma protein-free supernatant or a dilution of urine. A 10 micrometer muBondapak phenyl column with an eluting solvent of water--methanol--1 M phosphate buffer, pH 7 (70:30:0.5) was used, with UV detection of the effluent at 220 nm. Azidocillin potassium salt [potassium-6-(D-(-)-alpha-azidophenyacetamido)-penicillanate] was used as the internal standard and quantitation was based on peak height ratio of amdinocillin to that of the internal standard. The assay has a recovery of 74.4 +/- 6.3% (S.D.) in the concentration ranges of 0.1-20 microgram per 0.2 ml of plasma with a limit of detection equivalent to 0.5 microgram/ml plasma. The urine assay was validated over a concentration range of 0.025-5 mg/ml of urine, and has a limit of detection of 0.025 mg/ml (25 microgram/ml) using a 0.1-ml urine specimen per assay. The assay was applied to the determination of plasma and urine concentrations of amdinocillin following intravenous administration of a 10 mg/kg dose of amdinocillin to two human subjects. The HPLC and microbiological assays were shown to correlate well for these samples.