Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Biological samples”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 667 records · Page 37Linked to original sources
PubMed2004

Column-switching procedures for the fast analysis of drugs in biologic samples.

Liquid chromatography coupled with mass spectrometry in single and dual mode (LC-MS and LC-MS/MS) is the method of choice for the quantification of drugs and their metabolites in biologic fluids. Following the new challenges encountered in the process of drug development, liquid chromatography-mass spectrometry has been found to achieve high-throughput analysis. With this impressive tool, the sample preparation step before analysis is simplified, and the analytic process speeded up. Several generic approaches have recently been developed for the sample extraction coupled on line with a LC-MS system. In this paper, different extraction supports allowing the direct injection of biologic fluids were investigated, namely, restricted-access media, large-size particle, and monolithic phases. In the column-switching configuration, these supports, coupled with microbore analytic columns, were found suitable for the fast analysis (total analysis time of less than 10 minutes) of different drugs and their metabolites in biologic matrices at the nanogram per milliliter level.

Chromatography, Liquid↗
PubMed1996

Synthesis and characterization of polymer-coated mixed-functional stationary phases with several different hydrophobic groups for direct analysis of biological samples by liquid chromatography.

Three types of polymer-coated mixed-functional (PCMF) silica column-packing materials for the direct analysis of biological liquids have been synthesized. These packing materials all have polyoxyethylene groups as their hydrophilic part but differ in their hydrophobic part (methyl groups for Me-POE, phenyl groups for Ph-POE, and octyl groups for Oc-POE). Retention characteristics with respect to several drug molecules, and protein recovery, were studied for each of the three types with different amounts of hydrophobic groups attached. Of the PCMFs prepared, Oc-POE showed the greatest overall retention, and Me-POE displayed the greatest protein recovery.

Anti-Bacterial Agents↗
PubMed1986

A one-step enzymatic assay for the measurement of 17 beta-hydroxy- and 17-oxo-steroid profiles in biological samples.

We describe a simple enzymatic method for the sensitive and specific detection and quantitation of families of hydroxy- and oxo-steroids in biological mixtures. Analysis of the profiles of individual steroids may be achieved following their chromatographic separation. The objectives of this analytical system are, therefore, different from conventional methods which are designed to measure single steroids with a high degree of specificity. The method employs highly purified and active bacterial hydroxysteroid dehydrogenases (HSD) which promote stereospecific, nicotinamide nucleotide-dependent oxidations and reductions at specified positions of steroids. In the presence of catalytic quantities of steroids these enzymes promote the transfer of hydrogen (transhydrogenation) between NADH and NAD analogues. A recently purified 17 beta-HSD from an Alcaligenes species (D. W. Payne and P. Talalay, J. biol. Chem., 260, 13468-13655, 1985) shows almost complete specificity for the 17 beta-hydroxy- and 17-oxo-groups of both C18 and C19 steroids. This enzyme catalyzes steroid-dependent transhydrogenation between NADH and the thionicotinamide analogue of NAD (S-NAD). When these components are incubated at pH 8.5 in the presence of minute quantities of steroid substrates, S-NADH (measured at 398 nm where NADH does not absorb) accumulates at a constant rate which is proportional to the concentrations of steroid and enzyme. The linear increase in absorbance with time is a measure of the total concentration of 17 beta-hydroxy- and 17-oxo-steroids, and can be used to detect subpicomol quantities of steroids. The method is illustrated by the detection and identification of free and conjugated androgens in human serum following their separation by high pressure liquid chromatography. The specificity of the transhydrogenase assay is completely dependent on the specificity of the enzyme and is thus applicable to the detection of other hydroxy- and oxo-steroids by making use of HSDs with appropriate specificities (e.g. 3 alpha-HSD for the measurement of 3 alpha-hydroxy- and 3-oxo-steroids). The simple one-step reaction lends itself to automation, needs no auxiliary detection systems, and requires only an inexpensive colorimeter.

17-Ketosteroids↗
PubMed1998

Monoclonal antibody-based immunoassays for the specific quantitation of rat PAI-1 antigen and activity in biological samples.

Two enzyme-linked immunosorbent assays (ELISAs) for the quantitation of rat plasminogen activator inhibitor-1 (PAI-1) antigen and activity, respectively, in biological fluids were developed using monoclonal antibodies raised against recombinant rat PAI-1. These assays had a lower limit of sensitivity in plasma of 0.3 and 0.15 ng/ml, respectively. The intra-assay, inter-assay and inter-dilution coefficients of variation were 9, 14 and 9%, respectively, for the antigen assay and 8, 17 and 13%, respectively for the activity assay. Assay recoveries of recombinant rat PAI-1 (5 to 20 ng/ml) added to plasma were 73 to 88% and 89 to 93% for the antigen and the activity assay, respectively. The level of PAI-1 antigen in rat plasma was 1.8 +/- 0.9 ng/ml (mean +/- SD, n = 18), with a corresponding value of 1.0 +/- 0.5 ng/ml for PAI-1 activity. In lysed platelet-rich rat plasma PAI-1 antigen was 6.0 +/- 1.0 ng/ml (n = 8) and PAI-1 activity was 2.3 +/- 0.4 ng/ml (n = 8). Endotoxin injection (0.5 mg/kg) induced a time-dependent increase of both PAI-1 antigen and PAI-1 activity levels in rat plasma. eventually resulting in a 100- to 200-fold increase (p < 0.0001 vs. baseline). A linear correlation was found between PAI-1 antigen and activity levels in normal plasma (r = 0.63, n = 18, p < 0.01) and in plasma from endotoxin-treated rats (r = 0.90, n = 35, p < 0.001). Application of these assays for the analysis of gel filtration experiments of plasma from endotoxin-treated rats demonstrated that PAI-1 antigen eluted as two peaks (with corresponding Mr of approximately 430 kDa and 61 kDa) whereas PAI-1 activity eluted as a single peak corresponding with the high molecular weight antigen form. Thus, these unique assays allowing the specific determination of rat PAI-1 antigen and rat PAI-1 activity may constitute important tools for further investigations on the pathophysiological role of PAI-1 in a variety of experimental rat models.

Animals↗
PubMed2001

Fast clean-up for polychlorinated dibenzo-p-dioxins, dibenzofurans and coplanar polychlorinated biphenyls analysis of high-fat-content biological samples.

A fast clean-up procedure for the low level analysis of polychlorinated dibenzo-p-dioxins. polychlorinated dibenzofurans and coplanar polychlorinated biphenyls in highly fatty biological matrices using high capacity disposable multi-layer silica columns is presented. Results were compared with gel permeation chromatography for removal of lipids. Analytical criteria such as recovery rates, repeatability, reproducibility and robustness are evaluated through a broad range of biological matrices and reference materials analysis. The final proposed procedure for the complete analysis, including pressurized liquid extraction, Power-Prep system clean-up and GC-high-resolution MS analysis requires only 48 h, and allows the simultaneous preparation of up to 10 samples.

Benzofurans↗
PubMed1978

Determination of aluminum in biological samples by atomic absorption spectrophotometry with a graphite furnace.

Aluminum, generally considered non-essential and non-toxic, may accumulate in toxic amounts in the brain in cases of chronic renal failure. We describe a procedure for its analysis in biological fluids by atomic absorption spectrophotometry with a graphite furnace. No sample preparation is required and the procedure is sensitive at the appropriate concentrations. A sample of serum or urine is pipetted into the interior of the graphite tube, where it is sequentially dried, charred, and atomized. Precautions for sample handling are discussed and instrument settings are defined. Precision and accuracy of the method are evaluated, as are the effects of salts, protein content of serum, and specific gravity of urine. Serum of 23 persons who were not consuming aluminum-containing antacids contained 28 +/- 9 (SD) microgram of Al per liter (1.02 +/- 0.33 mumol/liter).

Aluminum↗
PubMed2006

Analysis of anatoxin-a in biological samples using liquid chromatography with fluorescence detection after solid phase extraction and solid phase microextraction.

Anatoxin-a is a naturally occurring, potent neurotoxin produced by some species of cyanobacteria in freshwaters. This toxin, which is a potential health hazard, especially to animals, has been determined in different biological matrices such as several cyanobacterial cultures and water samples and carps and mussels tissue using a sensitive High Performance Liquid Chromatography with Fluorescence detection method. Sonication was the technique selected for the extraction of intracellular anatoxin-a and solid phase extraction using weak cation exchange was used for the concentration and purification of the samples. 4-Fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) was used to convert anatoxin into a highly fluorescent derivative. Recovery experiments were performed for each type of matrix used in this work, and adequate values were obtained (71-87%). Limits of detection for anatoxin-a were estimated to be in the ng/L and ng/g level for water and cyanobacterial samples, respectively. The results obtained were also compared with those obtained after using solid phase microextraction, as an alternative for the extraction and purification of the samples. Advantages and disadvantages regarding to the efficiency for impurities removal, simplicity and rapidity and the potential for concentration enhancement of using both methodologies have been also discussed.

Chromatography, High Pressure Liquid↗
PubMed1997

Occurrence of 13(S)-hydroxyoctadecadienoic acid in biological samples.

The oxygenated metabolite of linoleic acid, 13(S)-hydroxyoctadecadienoic acid has recently been shown to play a role in cellular regulation. To detect this molecule in biological systems, we recently developed a specific polyclonal antibody. Using this antibody, we report the presence of 13(S)-hydroxyoctadecadienoic acid in human urine, cell culture media, and untreated goat serum for the first time by a specific, sensitive, and rapid enzyme immunoassay. Furthermore, the enzyme linked immunosorbent assay data are verified by gas chromatography/mass spectrometry analysis of the same samples.

Animals↗
PubMed2001

Multi-residue extraction-purification procedure for corticosteroids in biological samples for efficient control of their misuse in livestock production.

A fast and efficient multi-residue extraction-purification procedure was developed for 12 corticosteroids in biological matrices (hair, urine and meat), in order to control their illegal use as growth promoters in cattle. Detection and identification of the analytes were achieved using a previously described LC-MS-MS method based on negative electrospray ionisation and a triple quadrupole analyser. The presented procedures included acid (hair) or enzymatic (urine and meat) hydrolysis, C18 reversed-phase SPE, Na2CO3 liquid-liquid clean-up and SiOH normal-phase SPE. The detection limits of the developed methods were between 2.9 and 9.3 pg/mg (ppb) for hair samples and in the 40 - 70 pg/g (ppt) range for the urine or meat samples. The acid hydrolysis used for corticosteroid extraction in hair was optimised using an experimental design and response surface methodology. Achieved performances were linked to a physico-chemical approach based on the corticosteroids specific C17 side-chain. This original multi-residue and multi-matrices analytical methodology will be used for further metabolism studies.

Adrenal Cortex Hormones↗
PubMed1983

Enzymatic measurement of ethanol or NAD in acid extracts of biological samples.

An enzymatic method for the measurement of ethanol has been developed to permit analyses with unneutralized acid extracts of blood, liver, cell suspensions, or other biological materials. Components of the assay mixture include NAD, yeast alcohol dehydrogenase, tris(hydroxymethyl)aminomethane (Tris), and lysine. Tris is a trapping agent for the reaction product, acetaldehyde. Lysine is used to maintain the pH at 9.7 where oxidation of ethanol is quantitative and most rapid, even when as much as 0.2 ml of 0.5 N HClO4 is added. Lysine also causes the reaction to be 2 to 4 times faster than it is when either glycine or 2-amino-2-methyl-1-propanol is used as the buffer. The assay is linear up to an ethanol concentration of 0.125 mM in the reaction mixture and is complete by 4 min. By substituting ethanol for NAD in the reagents, the assay performs equally well in measuring NAD.

Alcohol Dehydrogenase↗
PubMed1985

High-performance liquid chromatographic determination of plasma and brain histamine without previous purification of biological samples: cation-exchange chromatography coupled with post-column derivatization fluorometry.

A highly sensitive and specific method was developed for the determination of histamine in biological materials by high-performance liquid chromatography with a cation exchanger and an automated Shore's fluorometric detection system. Since substances causing interference in Shore's o-phthalaldehyde method, such as ammonia, histidine, spermine and spermidine, were completely separated on the column and their fluorescent intensities were much less than that of histamine in this detection system, histamine could be determined by injecting a perchloric acid extract of human plasma or mouse brain tissue directly onto the column without any previous purification procedure. The lower limit of detection of histamine by this method is 0.05 pmol, and the within-day and day-to-day variations in plasma histamine assay are less than 3%. The plasma histamine level in normal human subjects was found to be 4.0 +/- 1.6 pmol/ml (mean +/- S.D., n = 20). A good linear correlation was obtained between values for the histamine contents of mouse brain tissues determined by this method and by a radioenzymatic method with a purified histamine-N-methyltransferase preparation. The histamine levels of whole brain, hypothalamus, thalamus, brain stem and frontal cortex in male ddY mice were 367 +/- 38, 1143 +/- 70, 414 +/- 66, 196 +/- 47 and 467 +/- 91 pmol/g of wet tissue (mean +/- S.D.), respectively.

Adult↗
PubMed2006

Three spectrophotometric assays for the measurement of the five respiratory chain complexes in minuscule biological samples.

BACKGROUND: The measurement of the activities of the five complexes comprising the respiratory chain has proven to be a major challenge when a limiting amount of biological material is available. Here we report a set of three convenient assays that allows this measurement under such circumstances. METHODS: One assay relies on the sequential addition of reagents to measure first complex IV activity, followed by complex II+III, and then glycerol-3-phosphate dehydrogenase+complex III activities and finally isolated complex III activity. A second assay measures the activity of complex II followed by glycerol-3-phosphate dehydrogenase and isocitrate dehydrogenase. A third assay measures rotenone-sensitive complex I activity and subsequently oligomycin-sensitive complex V activity. RESULTS: These assays have been successfully used on extracts of small numbers of human cells displaying various defects in the respiratory chain, and on frozen tissue homogenates of retina and very early mouse embryos. CONCLUSIONS: The strength of this set of assays lies both in its rapid and simple execution and its capacity for immediate detection of partial defects, because each activity can be compared with one or two other activities measured in the same sample.

Animals↗
PubMed1987

The determination of glutathione, cyst(e)ine, and other thiols and disulfides in biological samples using high-performance liquid chromatography with dual electrochemical detection.

A determination of glutathione, cysteine, and their disulfides using HPLC and dual electrochemical detection (HPLC-DEC) was described previously but was not validated in biological tissues for these and other important thiols and disulfides (SH/SS). Thus, our objectives were to develop this method to quantify simultaneously reduced and oxidized glutathione, cysteine, cystine, and other SH/SS in various tissues, including human blood and plasma, rat liver and hippocampus, mosquito, and spinach leaf. Optimal conditions were determined for sample processing and analysis using metaphosphoric acid and HPLC-DEC. Authentic standards of 10 common SH/SS compounds were resolved and eluted within 15 min, and all standard curves were linear from 5 to 1600 pmol. Validation was based on the following: First, tissue sample sizes were proportional to peak areas over an eightfold range. Second, recovery of SH/SS added to samples before processing was 96-101%. Finally, the results were equivalent and correlated highly with values for total SH by 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) assay (r2 = 0.996) and for total glutathione by DTNB-GSSG reductase assay (r2 = 0.998). The life span of the Au/Hg electrode was limited to 200-500 samples based on the lineal range of standard curves. On the basis of these results, we believe that this method will fill a long-time need for the simultaneous determination of SH/SS in biological tissues.

Aedes↗