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Comparative use of three electrokinetic capillary methods for the determination of drugs in body fluids. Prospects for rapid determination of intoxications.

Three electrokinetic capillary methods, micellar electrokinetic capillary chromatography, capillary zone electrophoresis and capillary isotachophoresis, are shown to be well suited for the rapid screening and confirmation of drugs in serum and urine of patients with medical drug overdoses (intoxications), situations where rapid identification without precise quantification is needed. Patients' samples obtained from the emergency care unit were analysed in an instrument featuring on-column, fast forward-scanning multi-wavelength detection and the data were compared with those obtained by conventional methods. The drugs studied included salicylate, acetaminophen (paracetamol) and antiepileptics. In cases with high drug concentrations, body fluids can be injected directly or may have only to be diluted (urine) or ultrafiltered (serum) prior to analysis, providing results within about 30 min. Thus, electrokinetic capillary methods can be employed for rapid drug screening, provided that instrumentation with a database for peak identification is available.

Acetaminophen↗

Separation of synthetic mixtures of sugars and of sugars in body fluids with a high-resolution carbohydrate analyzer.

With use of a specially designed gradient-generating system, of anion-exchange resins of 10-20 mum particle diameter, and of an orcinol solution in concentrated sulfuric acid (1 g/liter) as detection reagent, a mixture of 16 carbohydrates can be separated in less than 4 h. The limit of detection for most sugars is in the 0.1--2.0 nanomole range. The detection reagent is stable for months and does not become colored. The reagent mixes with the column effluent in an exothermic reaction and allows use of relatively short mixing coils. The analyzer is appropriate for investigating body fluids rapidly and with high sensitivity.

Chromatography, Ion Exchange↗

Preliminary beta-tricalcium phosphate coating prepared by discharging in a modified body fluid enhances collagen immobilization onto titanium.

Because of its excellent scaffold properties toward bone cells, collagen has been recognized as a promising extracellular matrix protein for surface modification of titanium implants. Hydroxyapatite (HA) coatings have been investigated as a preliminary coating for collagen immobilization on titanium implants. However, the composition of HA-collagen is recognized as being difficult, and while many studies have suggested that biodegradable beta-tricalcium phosphate (beta-TCP) has better osteoconductivity than HA, the efficiency of preliminary beta-TCP coating for collagen immobilization on titanium surfaces has yet to be evaluated. This investigation aimed to evaluate the applicability of HA and beta-TCP coatings, prepared by discharging in modified body fluids, as preliminary collagen coatings. To increase collagen induction on preliminary HA and beta-TCP coatings, we used a new cathodic polarization method. X-ray photoelectron spectroscopy revealed that the bonding strength between the collagen NH(+) amino groups of collagen and phosphate (PO(4) (3-)) was greater on the beta-TCP coating than the HA coating. The preliminary beta-TCP coating was tightly crosslinked with RCOO(-) carboxyl groups of the collagen molecules and showed high cellular responses, even in the early stage of cell cultivation. Thus, this coating was found to be more effective than HA as a preliminary coating for collagen immobilization on titanium implants.

Animals↗

Improved analysis of malondialdehyde in human body fluids.

The widely used TBA assay for lipid peroxidation was modified to minimize artefactual oxidative degradation of lipids during the assay. Formation of the TBA-MDA condensation product was studied with and without exclusion of oxygen, and the concentration effect of BHT addition was examined. Oxygen was depleted from the reaction mixture by extensive argon gassing. Exclusion of oxygen resulted in decreased TBARS production in plasma but not in standard solutions. High BHT concentrations resulted in a similar effect. At concentrations higher than 3 mmol/l BHT exclusion of oxygen had no additional effect. By measuring n-butanol extracts in a multititer plate reader this modified method was made suitable as a preliminary screening assay of human body fluids for lipid peroxidation.

Adult↗

Postmortem determination of concentrations of stress hormones in various body fluids--is there a dependency between adrenaline/noradrenaline quotient, cause of death and agony time?

To find out whether a certain cause of death or a certain length of an agonal period shows specific adrenaline or noradrenaline profiles, heart blood, femoral vein blood, liquor, urine and vitreous humour were taken from corpses (n = 98) at the Medical School Hannover, and noradrenaline and adrenaline were determined using high-performance liquid chromatography (HPLC). Corpses were classified according to the following five categories: short agony, long agony, state after hanging, state after asphyxiation and state after CPR with documented administration of epinephrine. Once results were collected the adrenaline/noradrenaline quotient was determined. It became clear that there were no significant differences regarding the concentration of adrenaline and noradrenaline in the various body fluids in relation to the above-mentioned categories. The means adrenaline/noradrenaline quotients in femoral vein blood were 0.21 +/- 0.29 for hanged persons, 0.38 +/- 0.47 for asphyxiated persons, 0.17 +/- 0.19 for those with short agony and 0.42 +/- 0.43 for those with long agony, significantly below 1 (p < 0.001; p = 0.001; p = 0.003). For condition after CPR we found an adrenaline/noradrenaline quotient of 2.81 +/- 5.8. In liquor the adrenaline/noradrenaline quotients for short agony was 0.17 +/- 0.17, for hanged persons 0.18 +/- 0.19 and for asphyxiated ones 0.30 +/- 0.38, significantly lower than 1 (p < 0.001). In urine the adrenaline/noradrenaline quotients for all categories are lower than 1 (p < 0.001); short agony (0.13 +/- 0.09), long agony (0.21 +/- 0.16), hanged (0.15 +/- 0.16), asphyxiated (0.14 +/- 0.08) and CPR (0.14 +/- 0.06). In vitreous humour the quotients for short agony (0.14 +/- 0.28), long agony (0.13 +/- 0.12), hanged (0.07 +/- 0.09) and asphyxiated (0.09 +/- 0.11) are lower than 1 (p < 0.001). The spread of data for the adrenaline/noradrenaline quotient did not allow for any conclusions about cause of death and length of agony in individual cases.

Asphyxia↗

Surface modifications of a glass and a glass-ceramic of the MgO-3CaO.P2O5-SiO2 system in a simulated body fluid.

A glass of nominal composition (wt%) 17.25 MgO-52.75 3CaO.P2O5-30 SiO2 and a glass-ceramic obtained from it showed surface modifications when immersed in an acellular medium having a composition similar to that of human blood plasma. A (Ca, P)-rich layer, with an approximate Ca/P atomic ratio of 1.7, identified as hydroxyapatite, developed on both samples. The precipitated film on the glassy sample was weakly bonded, whereas that formed on the glass-ceramic was strongly adherent. The apatite precipitated during the in vitro tests on both samples grew as a needle-like structure with crystals about 150-200 nm long and 50-70 nm thick, as measured on specimens soaked for 1 month in the simulated body fluid (SBF). The presence of calcium and phosphate ions in the SBF contributed to the precipitation of the (Ca, P)-rich layers on both specimens.

Binding Sites↗

Sequential changes in plasma renin activity and plasma catecholamines in mildly hypertensive patients during acute, furosemide-induced body-fluid loss.

To evaluate the role of adrenergic mechanisms in the acute response of renin to furosemide, plasma renin activity (PRA) and plasma catecholamine concentrations were measured for 3 h after i.v. administration of furosemide 1 mg/kg to 8 patients with mild essential hypertension. Furosemide induced a prompt and long-lasting increase in renin, with PRA more than doubled at all times. The increase in PRA within the first 30 min paralleled the peak increases in urinary water and sodium flow rates, and significant decreases in plasma volume and central venous pressure. There was no change in plasma catecholamine concentrations. Plasma noradrenaline was increased significantly at 60 min and adrenaline at 90 min, once furosemide had induced a marked loss of body-fluid and approximately 65% decrease in central venous pressure. Both catecholamines remained elevated until the end of the study, whereas urinary water and sodium flow rates had returned to their pre-treatment values by 150 min. Mean blood pressure was essentially unchanged throughout the study, whereas heart rate increased significantly after 90 min. The findings suggest that in mildly hypertensive patients adrenergic mechanisms are not involved in the initial renin response to furosemide, but they come into play later, probably as a result of reflex sympathetic activation triggered by marked volume depletion.

Adult↗

Total internal reflected resonance light scattering determination of chlortetracycline in body fluid with the complex cation of chlortetracycline-europium-trioctyl phosphine oxide at the water/tetrachloromethane interface.

A highly selective method of chlortetracycline (CTC) is proposed on the basis of the measurements of total internal reflected resonance light scattering (TMR-RLS) at water/tetrachloromethane (H20/CCl4) interfaces. In the pH range of 7.54-8.14, the interaction of the binary complex of Eu(III)/CTC in the presence of trioctyl phosphine oxide (TOPO) occurs at the H20/CCl4 interface, resulting in greatly enhanced TIR-RLS signals with the maximum peak located at 340 nm. The enhanced TIR-RLS intensity is in proportion to the CTC concentration in the range 0.98 to approximately 20.0 x 10(-7) mol/L. The limit of detection is 9.8 x 10(-9) mol/L. Synthetic samples and body fluid samples including human urine, human serum, and fresh milk were determined with the recovery of 95.4-106.4% and RSD of 2.9-3.9%.

Animals↗

Reversed-phase high-performance liquid chromatography of purine compounds for investigation of biomedical problems: application to different tissues and body fluids.

An overview of high-performance liquid chromatographic separation techniques (reversed-phase and ion-pair reversed-phase) used in the analysis of purine ribonucleotides, ribonucleosides and nucleobases, including procedures for sample preparation, is given. Coverage of the separation techniques is extended to the measurement of specific radioactivities of these compounds in tracer kinetic experiments for metabolic flux rate analyses. This article is focused on the development and adaptation of reversed-phase separation techniques for nucleotides, nucleosides and bases that are used to examine a variety of biomedical problems. The investigation of purine nucleotide metabolic disorders or physiological transitions in the pathomechanisms of different diseases and syndromes or in cell maturation processes, respectively, requires the application of chromatographic separation to a multitude of tissues and body fluids. These samples vary greatly in concentrations of purine compounds with low molecular mass, from ca. 5 mM to ca. 0.5 microM. The advantages and disadvantages of different techniques are critically discussed.

Animals↗