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PubMed1993

Strategies for the monitoring of drugs in body fluids by micellar electrokinetic capillary chromatography.

Electrokinetic capillary techniques can exploit numerous separation principles, making them flexible and easily applicable to a variety of separation problems. In recent publications, this emerging technology has been shown to be well suited for monitoring drugs and metabolites in body fluids, including serum, saliva and urine. Most attention has been focused on micellar electrokinetic capillary chromatography (MECC) because it permits the separation and determination of drugs with discrimination being largely based on differences in hydrophobicity. An overview of literature data on the MECC of drugs in body fluids and recent data obtained with antiepileptics in serum and saliva, with model mixtures of illicit drugs, and with extracts from urine specimens that tested positively for opiates and cocaine metabolites are presented. Emphasis is focused on buffer selection and simple sample preparation procedures, including direct injection of body fluids, ultrafiltration and solid-phase extraction.

Chromatography, Liquid↗
PubMed2000

Determination of paracetamol (acetaminophen) in different body fluids and organ samples after solid-phase extraction using HPLC and an immunological method.

A solid-phase extraction method routinely used for serum samples was improved and applied to the qualitative and quantitative determination of paracetamol in different body fluids, e.g. blood, urine, cerebrospinal fluid, synovial fluid, vitreous humor, and in tissue samples. A very simple method showed best results: Body fluids were mixed with phenacetine as internal standard and phosphate buffer (pH 6.8). Then protein was precipitated using acetonitrile. After strong centrifugation the supematant was transferred to a preconditioned Bakerbond C18-SPE-column. Elution with methanol without a prior washing step showed best recovery rates. The extracts were investigated using high-performance liquid chromatography with ultraviolet detection, a photometrical and an immunochemical method.

Acetaminophen↗
PubMed2006

Dysregulation of retinoid transporters expression in body fluids of schizophrenia patients.

This study aims to find the biomarkers or associated proteins in body fluids of schizophrenia patients so that we can further understand the etiology of schizophrenia. We applied proteomic technologies combining two-dimensional electrophoresis with Coomassie blue staining and mass spectrometry and identified a procedure for the clinical screening of disease-influenced body fluid proteins in two sets of samples, plasma from 19 schizophrenia patients and cerebrospinal fluid (CSF) from 35 drug-treated schizophrenic patients and 36 healthy controls. The expression of transthyretin (TTR) tetramer increased significantly in plasma of schizophrenic patients after a valid 2 months in-hospital antipsychotic treatment. Conversely, the expression of the TTR tetramer and apolipoprotein E (ApoE) was down-regulated by up to 1.68 and 3.62 times, respectively, in the CSF of schizophrenia patients compared to that of normal controls, which has not been reported previously. Considering that the TTR tetramer and ApoE are both retinoid transporters, retinoid dysfunction might be involved in the pathology of schizophrenia.

Adolescent↗
PubMed1993

Magnesium content of human myometrium and placenta during various stages of gestation, and of different body fluids at term.

Pregnancy is marked by a state of hypomagnesaemia but not much is known about the effects of maternal magnesium deprivation on the fetus. The aim of this study was therefore to measure magnesium concentrations in human myometrial and placental tissues and in different body fluids during pregnancy and at term, using atomic absorption spectrophotometry. The magnesium concentration in umbilical cord blood was higher than in the maternal blood, supporting the existence of an active magnesium transport system. There was also a difference between the magnesium concentration in the amniotic fluid and that in the umbilical cord blood. The magnesium content of myometrium increased from the 32nd week of pregnancy, reaching its maximum level at the 37th week of gestation. Later there was significant decrease in magnesium level until the end of pregnancy. The magnesium content of placental tissue did not change up to the 30th week of pregnancy; however, it then decreased continuously during the last trimester. The significant improvement experienced in certain pathological conditions of pregnancy treatment with magnesium supports the existence of a magnesium deficit induced by pregnancy and the need for magnesium supplementation.

Amniotic Fluid↗
PubMed1996

Capillary electrophoresis for drug analysis in body fluids.

Capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MECC) represent attractive methods for the determination of drugs and metabolites in body fluids. In CZE, minute (nanoliter) quantities of samples are applied to the beginning of a fused-silica capillary filled with buffer. On application of a high-voltage DC field, charged solutes begin to separate and are swept through the capillary by the combined action of electrophoresis and electroosmotic bulk flow and are on-column detected toward the capillary end. In MECC, the buffer contains charged micelles (e.g., dodecyl sulfate micelles) and both uncharged and charged solutes separate based on differential partitioning between the micelles and the surrounding buffer and, if charged, also by differential charge effects, including electrophoresis. Based on validated MECC drug assays developed in our laboratory, key aspects of measuring drug levels by MECC, including sample preparation, solute detection and identification, quantitation, reproducibility, and quality assurance are discussed. Drug levels determined by MECC are shown to be in good agreement with those obtained by nonisotopic immunoassays and/or high-performance liquid chromatography (HPLC). Using on-column multi-wavelength detection, this technology is also well suited for toxicological drug screening and confirmation and for the exploration of drug metabolism. Compared with HPLC and gas chromatography, capillary electrophoresis has distinct advantages, including automation, small sample size, minimal sample preparation, use of very small amounts of organic solvents and inexpensive chemicals, ease of buffer change and method development, and low cost of capillary columns. Electrokinetic capillary assays are complementary to the widely employed immunoassays. The state of the art and the pros and cons of capillary electrophoresis for the determination of drugs in body fluids are discussed with the goal of encouraging newcomers to start using this emerging analytical methodology.

Body Fluids↗
PubMed1989

The distribution and duration of hantaan virus in the body fluids of patients with hemorrhagic fever with renal syndrome.

The distribution and duration of Hantaan virus (HTNV) in the body fluids of patients were studied by immunofluorescence, reverse passive hemagglutination, and cell culture assays. Virus antigen of hemorrhagic fever with renal syndrome in peripheral blood mononuclear cells (PBMCs) was usually present before day 11 of the disease, especially from days 4-7. Virus isolates were more readily recovered from plasma early in the course of the illness and less frequently after day 7. The use of PBMCs rather than plasma enabled isolates to be recovered at a rate nearly twice that permitted by plasma and allowed the isolation peak of HTNV (days 4-7 after onset of disease) to extend an additional 2 or 3 d, thus prolonging the period of detectable viremia until days 8-11. PBMCs were especially useful in isolating viruses from patients with hemorrhagic fever with renal syndrome in whom antibody titers were generally high during the acute phase of the disease. HTNV was isolated from the cerebrospinal fluid of patients, but was difficult to recover from other body fluids.

Antibodies, Monoclonal↗
PubMed2002

Human cytomegalovirus load in various body fluids of congenitally infected newborns.

BACKGROUND: Congenital human cytomegalovirus (hCMV) infection is the most common intrauterine viral disease in western countries. Little is known about hCMV virus load in various body fluids of congenitally infected children. OBJECTIVES: To determine virus load in various body fluids. To assess the impact of hCMV virus load to predict the outcome of congenitally infected newborns and efficacy of antiviral therapy. STUDY DESIGN: Cord vein blood, urine, and cerebrospinal fluid (CSF) of congenitally hCMV-infected children were investigated and hCMV load was determined by quantitative polymerase chain reaction (PCR). Fourteen of 30 children had clinical symptoms and/or pathological laboratory results and 16 had none of them at birth. Ganciclovir was given to 21 children (10 of them with symptoms, 11 of them without symptoms). Viral load before and after therapy was measured. RESULTS: There was a significant difference between median virus load in cord vein blood (2.3 x 10(3) copies per ml) and in urine (4.2 x 10(5) copies per ml; P<0.001) at diagnosis of congenital hCMV infection. At that time, no significant difference of virus load was found between the various groups (symptomatic vs. asymptomatic; with therapy vs. without therapy), neither in serum nor in urine. Comparing median virus load in urine before (3.0 x 10(5) copies per ml) and after therapy (2.0 x 10(3) copies per ml), a significant decrease was observed (P<0.001). Virus load in CSF was always found to be less than 400 copies per ml, and only those children with symptoms showed a positive result. CONCLUSION: At birth, virus load in urine seems to be superior to that in cord vein blood to reflect the situation in the organs precisely. As predicting factor for the risk of developing symptoms, only hCMV detection in the CSF appears to be promising. The significant decrease of virus load in children with therapy may reflect the efficacy of therapy. Studies including a greater number of children are needed.

Antigens, Viral↗
PubMed2001

Sensitive determination of pethidine in body fluids by surface ionization organic mass spectrometry.

We have presented a simple and sensitive method for determining pethidine, a narcotic analgesic drug in body fluids by gas chromatography (GC)/surface ionization organic mass spectrometry (SIOMS). Good linearity was obtained in the range of 0.625-25 ng/ml of whole blood and urine by mass chromatography, and in the range of 0.05-2 ng/ml of whole blood by selected ion monitoring (SIM). Pethidine and diphenylpyraline (internal standard) were extracted from body fluids with Bond Elut Certify cartridges; their recoveries were above 95%. The detection limits (signal-to-noise ratio=3) were estimated to be 0.2 ng/ml of whole blood or urine by mass chromatography, 0.02 ng/ml of whole blood by SIM.

Analgesics, Opioid↗
PubMed2002

Nitric oxide synthase blockade and body fluid volumes.

The influence of chronic nitric oxide synthase inhibition with N G-nitro-L-arginine methyl ester (L-NAME) on body fluid distribution was studied in male Wistar rats weighing 260-340 g. Extracellular, interstitial and intracellular spaces, as well as plasma volume were measured after a three-week treatment with L-NAME (approximately 70 mg/kg per 24 h in drinking water). An increase in extracellular space (16.1 +/- 1.1 vs 13.7 +/- 0.6 ml/100 g in control group, N = 12, P<0.01), interstitial space (14.0 +/- 0.9 vs 9.7 +/- 0.6 ml/100 g in control group, P<0.001) and total water (68.7 +/- 3.9 vs 59.0 +/- 2.9 ml/100 g, P<0.001) was observed in the L-NAME group (N = 8). Plasma volume was lower in L-NAME-treated rats (2.8 +/- 0.2 ml/100 g) than in the control group (3.6 +/- 0.1 ml/100 g, P<0.001). Blood volume was also lower in L-NAME-treated rats (5.2 +/- 0.3 ml/100 g) than in the control group (7.2 +/- 0.3 ml/100 g, P<0.001). The increase in total ratio of kidney wet weight to body weight in the L-NAME group (903 +/- 31 vs 773 +/- 45 mg/100 g in control group, P<0.01) but not in total kidney water suggests that this experimental hypertension occurs with an increase in renal mass. The fact that the heart weight to body weight ratio and the total heart water remained constant indicates that, despite the presence of high blood pressure, no modification in cardiac mass occurred. These data show that L-NAME-induced hypertension causes alterations in body fluid distribution and in renal mass.

Animals↗
PubMed2003

Olfactory sensitivity of the gilthead seabream (Sparus auratus L) to conspecific body fluids.

The potential for intraspecific chemical communication in the gilthead seabream (a marine perciform) was investigated by assessing the olfactory sensitivity to conspecific body-fluids (water occupied by conspecifics, intestinal fluid, urine, semen, egg fluid) by multiunit electrophysiological recording from the olfactory nerve. The olfactory system was responsive to water previously occupied by conspecifics, and the active compound(s) could be extracted by solid-phase extraction. The olfactory system was extremely sensitive to body fluids of sexually mature conspecifics: thresholds of detection were 1:10(7.4) (intestinal fluid), 1:10(6.1) (gametes), and 1:10(4.2) (urine). The olfactory system was also sensitive to amino acids with thresholds of detection from 10(-8.1) M (L-leucine) to 10(-6.1) M (L-phenylalanine). However, a range of other known fish odorants (steroids, bile acids, and prostaglandins) failed to evoke significant responses. Given the high olfactory sensitivity to intestinal fluid and the low urine release rates of marine compared with freshwater fish, we suggest that chemical communication is likely to be mediated via compounds present in the intestinal fluid rather than urine. Furthermore, the types of chemicals involved are likely to be different from those of freshwater fish. Their exact chemical identity and biological roles remain to be established.

Animal Communication↗
PubMed2001

Quantitative detection of Toxoplasma gondii DNA in human body fluids by TaqMan polymerase chain reaction.

OBJECTIVE: A new quantitative polymerase chain reaction (real-time PCR) was designed to detect Toxoplasma DNA in human body fluid samples. METHODS: Real-time fluorescence detection of amplification product formation on the basis of the TaqMan-System was established with Toxoplasma 18S rDNA as a target gene. RESULTS: The method provides a high sensitivity comparable to conventional nested PCR procedures and generates quantitative data when detecting toxoplasmic DNA in human blood, cerebrospinal or amniotic fluid. Moreover, data were obtained investigating blood samples from an immunocompromised patient with reactivated toxoplasmosis after allogeneic bone marrow transplantation, monitoring the therapeutic effect. CONCLUSIONS: The potential application of this method to detect Toxoplasma DNA in body fluids and to follow the development of parasitemia under therapy could be demonstrated.

Animals↗
PubMed1999

[Changes in body fluid compartments estimated by multifrequency bioelectric impedance analysis in patients treated by hemodialysis].

Bioimpedance is a simple non-invasive method of assessing body composition. The aim of this study was to investigate the effect of changes in body fluid related to haemodialysis on measured bioimpedance parameters, to determine correlation between bioimpedance data and the volume of ultrafiltratum, and to collect data on alterations of body fluids resulting from a treatment. Measurements were done on 19 patients (mean age 36.7 years) prior to and after haemodialysis. After dialysis we found significantly higher impedance values on each measured frequency (1, 5, 10, 50, 100 kHz) (p < 0.001). An inverse correlation was found between the changes in body weight (x) and resistance at 50 kHz (y): y = 8.4830-2.1850x (r = 0.7167, p < 0.01). Total body water calculated by bioimpedance analysis (BIA) decreased from 38.47 +/- 8.567 litres (56.07%) to 35.06 +/- 8.045 significantly (p < 0.001), and the reduction of extracellular water proved to be also significant [from 15.76 +/- 2.992 litres (22.97%) to 14.06 +/- 2.736 litres (21.17%), p < 0.001]. The relationship between the change in calculated body water (x) and the volume of ultrafiltratum (y) is: y = -0.5590 + 0.5864x (r = 0.4898 p < 0.05), bioimpedance in our study overestimated the fluid loss by 55%. The intradialytical shifts between extra- and intracellular spaces might be responsible for the difference in the two values.

Body Fluids↗
PubMed1981

Ionized calcium in body fluids.

This article will review the methods currently available to the clinician and research worker for measuring the concentration of ionized calcium in various body fluids including whole blood, serum, plasma, urine, cerebrospinal fluid, milk, and synovial fluid. The methods to be reviewed are based on procedures involving bioluminescence, colorimetry and ion-selective electrodes. Emphasis will be given to the precision and, wherever possible, accuracy of each technique. Possible sources of error and interfering agents will be identified. Attention will be given to the recommended conditions for measuring ionized calcium in each body fluid. An assessment will be made of the theoretical and practical importance of measuring ionized calcium rather than total calcium and of its value in clinical medicine.

Adult↗
PubMed2001

Proteomic profiling from human samples: the body fluid alternative.

Proteomics is one of the technologies rapidly changing our approach to drug development. The applications of proteomics, particularly with reference to analysis of body fluid samples, will be described. Proteomic analysis involves the systematic separation, identification and characterisation of proteins present in a biological sample. By comparing the proteins present in diseased samples with those present in normal samples, it is possible to identify changes in expression of proteins that potentially may be related to organ toxicity. Proteomics is regarded as a sister technology to genomics. Although the pattern of gene activity will be abnormal in a tissue with a pathological lesion, there can be a poor correlation between the level of activity of different genes and the relative abundance within the tissue of the corresponding proteins. This is especially true where the mode of action of the test material interferes with protein synthesis and/or post translational modification. Consequently, the information about a pathological process that can be derived at the level of gene activity is incomplete. Proteomics has now made it possible to analyse proteins using high throughput, automated techniques. Although both mRNA and proteomic profiling can be applied to tissue samples, analysis of body fluids (e.g. serum, urine, CSF, synovial fluid) is restricted to proteomics. In these cases the protein composition is derived from many tissues and processes. Proteomic analysis can yield information on disease processes and potential response to treatment. Examples will be presented of the identification of surrogate markers for hepatocellular carcinoma, breast cancer, from cerebrospinal fluid in humans and gentamicin toxicity in the rat.

Body Fluids↗
PubMed1996

Hematologic and body fluid changes during simulated high altitude exposure in naproxen-treated rats.

Arterial blood oxygen saturation, body fluid and hematological parameters were studied in control, naproxen (a prostaglandin synthesis inhibitor)-treated control, altitude-exposed, and naproxen-treated altitude-exposed rats after intermittent exposure of 8 h/d for 6 consecutive days to a simulated high altitude of 6,100 m (barometric pressure 349 +/- 3 mmHg). Arterial blood oxygen saturation was reduced in altitude-exposed rats, but increased significantly to near control level in naproxen-treated rats. On the other hand, 2,3-diphosphoglycerate in erythrocytes increased in altitude-exposed rats, but naproxen prevented this increase. The red blood cell count, hemoglobin concentration and hematocrit ratio were reduced significantly in drug-treated altitude-exposed rats when compared to the altitude-exposed group (without drug). Red cell mass was increased in the altitude-exposed group in comparison with control. Both red cell mass and mean corpuscular volume of altitude-exposed rats were shifted towards the control value when they were treated with naproxen. Drug-treated high altitude-exposure reduced the thiocyanate space, intracellular fluid volume and total body water content compared to drug-treated control rats. On the other hand, thiocyanate space and total body water content were increased significantly in drug-treated control rats in comparison with the control group.

Altitude↗
PubMed1977

Body fluid compartments.

The terms mole, molality, molarity, osmole, osmolality, osmolarity, osmolar gap and anion gap are defined and their clinical usefulness indicated. The following body fluid compartments are described: total body water (TBW), extracellular fluid (ECF), intracellular fluid (ICF), transcellular fluid TCF), plasma volume, red cell volume and interstitial fluid volume. Isotope-dilution techniques are briefly discussed and representative normal values for the various compartments according to sex and age are indicated. The physiological mechanisms that maintain the distinctive ionic compositions of the various fluid spaces are briefly outlined. New concepts of the function of the gel matrix and of the lymph drainage of the interstitium are presented. Opposing models to the sodium-potassium membrane pump are briefly described.

Body Fluid Compartments↗