High-performance liquid chromatographic analysis of hippuric acid in human blood plasma

A method has been developed for the isocratic high-performance liquid chromatographic analysis of hippuric acid in human blood plasma. After the addition of an internal standard (3-methoxysalicylic acid), plasma samples (1 ml) were made alkaline and extracted stepwise with methylene chloride and ethyl acetate. The detection limit was 50 pmol of hippuric acid per ml of plasma. The concentrations of hippuric acid in plasma from house painters (n = 8), with long-term exposure to solvent vapours from alkyd paints, were in the range 1-21 nmol/mol (median 11 nmol/ml). These values were statistically significantly higher than those for controls (n = 9): 2-8 nmol/ml (median 3 nmol/ml).     

High-performance liquid chromatographic analysis of isoxicam in human plasma and urine

A sensitive, selective, and rapid high-performance liquid chromatographic procedure was developed for the determination of isoxicam in human plasma and urine. Acidified plasma or urine were extracted with toluene. Portions of the organic extract were evaporated to dryness, the residue dissolved in tetrahydrofuran (plasma) or acetonitrile (urine) and chromatographed on a mu Bondapak C18 column preceded by a 4-5 cm X 2 mm I.D. column packed with Corasil C18. Quantitation was obtained by UV spectrometry at 320 nm. Linearity in plasma ranged from 0.2 to 10 micrograms/ml. Recoveries from plasma samples seeded with 1.8, 4 and 8 micrograms/ml isoxicam were 1.86 +/- 0.077, 4.10 +/- 0.107 and 8.43 +/- 0.154 micrograms/ml with relative standard deviations of 3.3%, 2.5% and 5.4%, respectively. The linearity in urine ranged from 0.125 to 2 micrograms/ml. The precision of the method was 3.3-9.0% relative standard deviation over the linear range.     

High-performance liquid chromatographic analysis of dinitrobenzene isomers

Summary An efficient, reproducible and rapid high-performance liquid chromatographic method, in normal phase mode, for the analysis of the three dinitrobenzene isomers is described. The method affords good linearity for each isomer in the range 10–160 g ml^–1. The total analysis time is only 10 minutes, and the method shows an accuracy of ±1.25% with a coefficient of variation from 0.30% to 2.85% for different levels of the dinitrobenzene isomers.     

High-performance liquid chromatographic analysis of steroid hormones

In the present work, a practical, rapid, reliable and isocratic reversed-phase high-performance liquid chromatographic (HPLC) method is described for the qualitative and quantitative analysis of estriol, estradiol-17 beta, estrone, testosterone, and progesterone. Chromatographic separation is complete in 16 min using a mobile phase of 50% acetonitrile (v/v) in water. The order of elution is estriol, testosterone, estradiol-17 beta, estrone, and progesterone; retention times are 2.5, 5.5, 5.6, 6.9, 16.3 min, respectively. Absorbance maxima of individual steroids is the limiting factor in quantitative determination. The recommended wavelengths for UV monitoring are E3 214, E2 280, T 254, E1 214, and P4 254 nm.     

High-performance liquid chromatographic analysis of azlocillin in serum

Summary A rapid and sensitive high performance liquid chromatographic method is described for the determination of azlocillin in serum. This method involves a short manual protein precipitation of the sample followed by an injection into a PR 18 column for separation and quantitation. The mobile phase was a 22% (V/V) solution acetonitrile in phosphate buffer pH 4.8 at a flow rate of 2,5 ml/min. The spectrophotometer detector was set at 220 nm with a sensitivity of 0.08 AUFS.     

High-performance liquid chromatographic analysis of dehydroepiandrosterone.

Qualitative and quantitative analysis of dehydroepiandrosterone and its conjugates in biological matrices and establishment of their relationships with physiological functions is a very active field. This review article discusses methods of separation and quantification of dehydroepiandrosterone and its conjugates using high-performance liquid chromatographic techniques.     

High-performance liquid chromatographic determination of benzodiazepines in human plasma

A simple and sensitive method for determination of benzodiazepines in plasma has been developed using high-performance liquid chromatography in a reverse-phase mode. The method is illustrated by application to plasma samples containing diazepam and N-desmethyldiazepam at concentrations which would be encountered during therapy, with limits of detection of 10 ng/ml and 2 ng/ml for diazepam and N-desmethyldiazepam, respectively.     

High-performance liquid chromatographic determination of bromazepam in human plasma

An assay using high-performance liquid chromatography has been developed for the determination of bromazepam in plasma. After a single-step extraction from basified samples with dichloromethane, using decarboxyloflazepate as an internal standard, samples were analysed using a reversed-phase Nova Pak 5-microns column with a mobile phase of methanol - phosphate buffer (60 + 40) adjusted to pH 7.6. The drugs were detected at 239 nm and the limit of detection was found to be 3 micrograms l-1 for bromazepam. The method is simple, rapid and sensitive and permits bromazepam levels in clinical and pharmacokinetic studies to be monitored.     

High-performance liquid chromatographic determination of arecoline in human saliva

Arecoline (methyl-1,2,5,6-tetrahydro-1-methyl nicotinate) is an alkaloid found in the areca catechu nut which is a major component of the 'betel quid' chewed by a large proporation of the population in India, South Asia and the South Pacific islands. It is commonly associated with the development of oral leukoplakia, oral submucous fibrosis and oral cancer. We have developed a new ion-pairing reversed-phase high-performance liquid chromatographic (HPLC) method for the determination of arecoline in saliva, using arecaidine (1,2,5,6-tetrahydro-1-methylnicotinic acid) as an internal standard. The optimal wavelength was established using UV absorbance scans. It was showed that 215 nm is the optimal wavelength to maximise the signal in detecting arecoline in the mobile phase. Arecoline was extracted from saliva with hexane-isoamyl alcohol (1%) and reconstituted with mobile phase for HPLC analysis. The developed method is an easy and reliable method of determining arecoline concentrations in saliva. Sensitivity, specificity, precision, accuracy and reproducibility of the method were demonstrated to be satisfactory for measuring the arecoline level.