High-performance liquid chromatographic assay for amiloride in plasma and urine

A sensitive and simplified high-performance liquid chromatographic procedure has been developed for quantification of amiloride in rabbit plasma, as well as human plasma and urine. Following protein precipitation with perchloric acid, the supernatant was directly injected into a C18 Nucleosil column. The mobile phase consisted of methanol-water (45:55) containing 0.1 M perchloric acid, and the compound was quantitated using a fluorescence detector at excitation and emission wavelengths of 286 and 418 nm, respectively. The average recovery was 97.6%. The calibration curve was linear over the range 2.0-20.0 ng/ml. The limit of detection was 0.5 ng/ml.     

High-performance liquid chromatographic analysis of amiloride in plasma and urine

A high-performance liquid chromatographic method has been developed for amiloride in rabbit plasma and urine which uses a reversed-phase C18 column, a mobile phase (flow-rate 2 ml/min) consisting of 32% acetonitrile in 0.15 M perchloric acid, pH 2.2, and spectrofluorometric detection via excitation at 286 nm. A simple extraction step with ethyl acetate eliminates interfering peaks. Short retention times of about 2.3 and 3.8 min are observed for amiloride and the internal standard, triamterene, respectively. The method can measure 4 ng/ml amiloride in plasma. This assay has been used to explore the pharmacokinetics of amiloride in rabbits. The plasma disposition profile is biexponential after a 50-mg intravenous bolus dose and there is no evidence for saturable elimination at zero-order infusion rates of 1.8, 3.6 and 7.2 mg/h.     

High-performance liquid chromatographic method using ultraviolet detection for measuring metrifonate and dichlorvos levels in human plasma.

We report high-performance liquid chromatographic methods using ultraviolet detection, developed for the first time in our laboratory with sensitivity to detect clinically significant concentrations of metrifonate (MTF), an experimental drug for Alzheimer disease, and its active anticholinesterase metabolite, dichlorvos (DDVP). The determination limit of the method for MTF and DDVP was 1 microgram/ml and 40 ng/ml, respectively. Stability of MTF and DDVP at various temperatures in water, buffered solutions and in human plasma were also studied.     

High-performance liquid chromatographic method for the determination of spectinomycin in turkey plasma.

A selective high-performance liquid chromatographic (HPLC) method with ultraviolet-visible (UV-VIS) detection was developed to measure therapeutic concentrations of spectinomycin in turkey plasma. Treatment of plasma samples with 3% trifluoroacetic acid in acetonitrile facilitated spectinomycin extraction and protein precipitation. After centrifugation, the stable derivatization reagent, 2,4-dinitrophenyl-hydrazine, was added to an aliquot of the supernatant, and the mixture was incubated for 30 min at 70 degrees C. Excess reagent was quenched with acetone and additional heating. The resulting derivative, a proposed spectinomycin-hydrazone, was separated from other compounds by reversed-phase HPLC during a short gradient run. The absorbance of the effluent was monitored spectrophotometrically with the UV-VIS detector set at 205 nm. The detector response was linear through the range of interest, 2-100 micrograms/ml.     

High-performance liquid chromatographic method for the determination of RGH-5702 in plasma samples.

A quick and selective high-performance liquid chromatographic method has been developed for the determination of RGH-5702 in plasma samples. A simple one-step extraction is used followed by reversed-phase chromatography and UV detection. This method allowed the separation of the compound and internal standard within 7 minutes. Validation of the method was performed prior to the assay of samples and continued throughout the study. Acceptable accuracy and precision was achieved at all concentrations investigated. The quantitation limit was 20 ng/ml using 1 ml of plasma. The method has been applied to the analysis of plasma samples from toxicokinetic studies in dogs.     

High-performance liquid chromatographic method for the determination of nefazodone and its metabolites in human plasma using laboratory robotics.

A quantitative analytical method, using high-performance liquid chromatography and ultraviolet detection, has been established for the determination of nefazodone (NEF) and its metabolites, m-chlorophenylpiperazine (mCPP),p-hydroxynefazodone (PHN), and hydroxynefazodone (HO-NEF), in human plasma. The fully automated, robotic procedure consisted of addition of internal standard (aprindine), extraction with butyl chloride, followed by phase separation, organic phase evaporation, reconstitution of the residue, and injection onto the chromatographic system. The limits of detection for NEF, mCPP, PHN, and HO-NEF were 5, 1, 10, and 5 ng/ml, respectively, at a signal-to-noise ratio of 4. The method had a linear range of 10-1000 ng/ml for NEF and HO-NEF, 20-2000 ng/ml for PHN, and 2.5-250 ng/ml for mCPP. Correlation coefficients of 0.996 or greater were obtained during validation and study sample analysis.     

High-performance liquid chromatographic method for the determination of pindolol in human plasma

A sensitive, simple and highly reliable high-performance liquid chromatographic method using fluorescence detection is reported for the determination of pindolol in plasma. This method involves a single extraction of pindolol from alkalinized plasma into methyl tert.-butyl ether followed by a back-extraction into dilute hydrochloric acid. Injection of the dilute acid phase directly onto an octyl (LC-8) bonded-phase column provides the final separation, and detection of pindolol is achieved by monitoring the intrinsic fluorescence of pindolol at 315 nm following excitation at 255 nm. The method is sensitive enough to measure with confidence pindolol plasma concentrations of 2 ng/ml using a 2-ml sample. No internal standard is required. This method has been applied to the analysis of 1500 human plasma samples by two different laboratories.     

High-performance liquid chromatographic determination of pilocarpine in plasma.

A high-performance liquid chromatographic procedure requiring neither derivatization nor complex sample work-up is reported for reproducibly and sensitively determining pilocarpine in plasma. Following stabilization of pilocarpine against in vitro hydrolysis using sodium fluoride, plasma samples were extracted and the extracts chromatographed on a 5-microns, low-carbon-load (6%) C18 reversed-phase column. The assay was linear between 10 and 300 ng/ml (r = 0.998). It had sufficient sensitivity to quantitate pilocarpine at concentrations as low as 10 ng/ml (signal-to-noise ratio > or = 4) using a 500-microliters sample. The assay appears to be the first published specifically for plasma determinations and has proven capable of supporting pharmacokinetics studies of pilocarpine disposition in the anesthetized dog.     

High-performance liquid chromatographic determination of ganciclovir in plasma.

A rapid, selective and sensitive isocratic reversed-phase high-performance liquid chromatographic method for the determination of ganciclovir in plasma samples was developed. This method, which was applied to the analysis of plasma ganciclovir from heart transplant patients under ganciclovir therapy for cytomegalovirus infections, represents a suitable analytical tool for drug monitoring and pharmacokinetic investigations.     

High-performance liquid chromatographic method for simple and rapid determination of linezolid in human plasma

A simple high-performance liquid chromatographic (HPLC) method was developed and validated for rapid quantification of linezolid in human plasma. Protein precipitation using a mixture of 5% trichloroacetic acid and methanol (3:1, v/v) provided a straightforward method of sample preparation and the internal standard eperezolid was employed. A concentration range from 0.20 to 40.0 mg/L was utilized to construct calibration curves, and analysis of low- (0.40 mg/L), medium- (7.50 mg/L) and high-quality (25.0 mg/L) control samples revealed excellent reproducibility (相似文献   

High-performance liquid chromatographic method for simultaneous determination of honokiol and magnolol in rat plasma

A high-performance liquid chromatographic (HPLC) method is described for the simultaneous determination of honokiol and magnolol in rat plasma. The plasma was deproteinized with acetonitrile which contained an internal standard (diphenyl) and was separated from the aqueous layer by adding sodium chloride. Honokiol and magnolol are extracted into the acetonitrile layer with high yield, and determined by reversed-phase HPLC and ultraviolet detection. The limits of quantitation for honokiol and magnolol were 13 and 25 ng ml⁻¹ in plasma, respectively, and recovery of both analytes was greater than 93%. The assay was linear from 20 to 200 ng ml⁻¹ for honokiol and from 40 to 400 ng ml⁻¹ for magnolol. Variation over the range of the standard curve was less than 15%. The method was used to determine the concentration-time profiles of honokiol and magnolol in the plasma following rectal administration of Houpo extract at a dose of 245 mg kg⁻¹, equivalent to 13.5, 24.4 mg kg⁻¹ of honokiol and magnolol, respectively.