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At least 235 records · Page 13Linked to original sources
PubMed2002

Application of isotopically labeled methylmercury for isotope dilution analysis of biological samples using gas chromatography/ICPMS.

An isotope dilution (ID) procedure for the determination of methylmercury (MMHg) in biological samples using an inductively coupled plasma mass spectrometer as detector after the capillary gas chromatographic separation (CGC/ICPMS) has been developed. For the first time, open-focused-microwave pretreatment has been used in conjunction with ID. Optimum conditions for the measurement of isotope ratios on the fast transient chromatographic peaks have been established. Mass bias was found to be about 1.5%/mass unit and was corrected by using the simultaneously measured thallium signals at 203Tl and 205Tl. After mass-bias correction, deviation of the theoretical mercury ratio values was found to be as low as 0.2%. Isotope ratio precisions based on the peak areas measurements were 0.3% RSD for 20 pg injected (as Hg absolute). The absolute detection limits were in the range of 20-30 fg for 202Hg and 201Hg. Methylmercury enriched in 201Hg has been synthesized by direct reaction with methylcobalamine. The concentration of the MMHg spike has been measured by reverse isotope dilution with a natural MMHg standard. The capabilities of CGC/ICPMS to measure isotope ratios were used to optimize sample derivatization by aqueous ethylation with NaBEt4 with respect to MMHg degradation pathways and quantitative recovery. The accuracy of the method developed has been validated with biological certified reference materials (CRM-463, DORM-1).

Animals↗
PubMed2005

Polymerase chain reaction for detection of Toxoplasma gondii in human biological samples.

Using the polymerase chain reaction (PCR), Toxoplasma gondii from gene TGR1E with primers TGR1E-1, TGR1E-2 (standard PCR), and from B1 gene with primers TM1, TM2, TM3 (hemi-nested PCR) was detected in biological samples from 347 individuals (441 biological materials). Of the total of 441 biological materials, T. gondii DNA was detected in 5.2 %; it was positive in the following samples: blood (n = 6), blood from newborns (2), biopsies (2) and samples of progenitor cells (2) (from candidates for bone marrow transplantation). DNA of T. gondii was also revealed in 11 samples (8.3 %) of 120 cases of pregnant women during prenatal examinations. A positive result in the blood was also found in two cases of newborn babies from mothers who were infected in later pregnancy. The positive PCR examination was confirmed by serological methods (ELISA and complement fixation test). Agreement of PCR results and the detection of antibodies against toxoplasma was found in 83.3 %. Rapid PCR examination for the confirmation of acute parasitemia T. gondii is particularly important for the patients in whom the infection may cause serious consequences (e.g., for fetus in pregnant women or for patients suffering from imunosuppression).

Adult↗
PubMed1984

Colorimetric estimation of inorganic phosphate in colored and/or turbid biological samples: assay of phosphohydrolases.

A simple method of inorganic phosphate determination for colored and/or turbid biological samples is described. The procedure is mild, and so is suitable for routine phosphohydrolase assays. Following deproteinization by ice-cold trichloroacetic (or silicotungstic) acid, the sample was treated with acid-washed charcoal to remove interference due to color. The phosphate in the colorless supernatant was assayed either by measuring the phosphomolybdate spectrophotometrically at 310 nm, following its extraction in organic solvents or by a modified Fiske and Subbarow method. The turbidity interference in the latter case was eliminated either by centrifugation, by sodium dodecyl sulfate treatment, or by extraction of reduced phosphomolybdate blue color by cyclohexanone. Though deproteinization by silicotungstic acid eliminated the turbidity problem, its use in conjunction with charcoal treatment was not convenient.

Animals↗
PubMed1991

Simultaneous high-performance liquid chromatographic determination of ascorbic acid and dehydroascorbic acid in biological samples.

The ascorbic acid (AA)-dehydroascorbic acid redox couple is an important component of many biological systems, and various physiological roles have been described for this vitamin. Simultaneous measurement of both AA and dehydroascorbate using high-performance liquid chromatography (HPLC) has proven difficult owing to detection problems. A simple, single-step HPLC assay for the simultaneous detection of both AA and dehydroascorbate was developed without the burden of derivatization of either compounds. This has proven to be a reliable technique and should be applicable to a wide variety of biological samples.

Animals↗
PubMed1994

Measurement of nitrite and nitrate levels in biological samples by capillary electrophoresis.

Nitrite is one of the products of NO-synthase in biological media. It is slowly oxidized in animals to nitrate. We developed a simple and rapid method to determine simultaneously nitrite and nitrate in biological samples. Capillary ion electrophoresis with direct UV detection at 214 nm was used employing a carrier electrolyte consisting of 10 mM sodium sulfate and an electroosmotic flow modifier. The detection limit in ultrafiltrates of plasma, urine and brain tissue extracts was 25 ng/ml for both compounds. Nitrate levels in human plasma and urine were in the microgram/ml range. Nitrite could not be detected. Rat brain tissue extracts contained detectable amounts of nitrite and nitrate.

Animals↗
PubMed2005

Development of a validated HPLC method for the determination of iodotyrosines and iodothyronines in pharmaceuticals and biological samples using solid phase extraction.

Identification, separation and quantitation of iodoaminoacids, is essential for the biological research and the clinical diagnosis of thyroid gland disease. Under this aspect a reversed-phase high-performance liquid chromatographic method was developed for the determination of thyroid gland hormones and some of their primary metabolites, 3,3',5,5'-tetra-iodo-L-thyronine (L-thyroxine), 3,3',5-tri-iodo-L-thyronine, 3,5-di-iodo-L-thyronine, L-thyronine, 3,5-di-iodo-L-tyrosine, 3-iodo-L-tyrosine and l-tyrosine. Analysis was performed on an Inertsil C(18) column with photodiode-array detection, using a 25 min gradient scale program of a binary mobile phase consisted of 0.1% aqueous solution of trifluoroacetic acid at pH 3 as solvent A and acetonitrile as solvent B, at a flow rate of 1 mL/min. Quantitation was performed using were obtained using theophylline as internal standard. The method was applied to commercial pharmaceuticals and biological samples (serum, urine and tissue). Drug-free urine and serum samples were spiked with known concentrations of the analytes standards and pretreated by solid phase extraction to remove matrix interferences. C(18) cartridges were used, yielding recoveries ranging from 87.1% to 107.6% for serum samples and from 92.1% to 98.7% for urine samples. With regard to total-T(4) concentrations in serum samples, results are cross-validated with RIA and found to agree well.

Chromatography, High Pressure Liquid↗
PubMed1996

Micellar electrokinetic chromatography separations and analyses of biological samples on a cyclic planar microstructure.

Micellar electrokinetic capillary chromatography (MECC) separations and analyses of biological samples on a planar glass microchip capillary electrophoresis device with laser-induced fluorescence solute detection are discussed. A cyclic channel system which permits dead volume free repeated column switching and thus the use of various channel lengths together with a relatively low applied separation voltage is described. It features an unbiased, dead volume free electrokinetic sample inlet system of approximately 12 pL. Because of the small cross section and favorable heat dissipation in glass microstructures, MECC separations with an electric field strength of up to 2000 V/cm achieving efficiencies of submicrometer plate heights can be performed. After a separation length of 2 cm, six fluorescein isothiocyanate labeled amino acids are shown to be separable within a few seconds and with an imprecision for peak areas (or heights) and detection times of < 2% and < 0.5%, respectively. Without application of electrokinetic solute stacking, the detection limit of fluorescein isothiocyanate labeled arginine is 3.3 nM, corresponding to approximately 40 zmol injected. Furthermore, the feasibility of directly applying human urine and serum samples onto the uncoated channel system is demonstrated and first data of the successful performance of a chip-based MECC immunoassay for serum theophylline are presented. Compared to MECC in conventional fused-silica capillaries, MECC analyses on microchips can be performed 1-2 orders of magnitude faster, with higher efficiency and at no expense of accuracy and precision. Furthermore, versatility is shown to be much increased with the use of a cyclic rather than a single-path channel system. The MECC separation efficiency of fluorescein isothiocyanate labeled amino acids is shown to be comparable to that obtained by gel electrophoresis performed in the same chip layout.

Amino Acids↗
PubMed1982

Trace elements analysis in biological samples by proton nuclear activation.

A method for the multielemental analysis of trace elements in biological samples by medium energy proton nuclear activation (PNA) has been developed. The response linearity, reproducibility and reliability of the method have been tested by a set of preliminary measurements. Results concerning the quantitative determination of the concentrations of Sr, Cu, Fe, Zn and Se in human serum are presented. Activation was induced with a 23 MeV proton beam from the AVF cyclotron of the University of Milan, by means of (p, 2n) reactions on the nuclei of the elements under study. The quantitative determination was obtained by an internal reference method and by comparison with a standard sample of serum doped with known quantities of the elements of interest. A clinical investigation of the zinc content of serum in children has been performed.

Activation Analysis↗
PubMed1966

Lead-210 and polonium-210 in biological samples from Alaska.

The naturally occurring concentrations of lead-210 and polonium-210 in certain biological samples from Alaska are unusually high. The concentration processes are similar to those observed for artificially produced radioactive fallout. Concentrations of these nuclides are greater in Alaskan natives than they are in other United States residents.

Alaska↗
PubMed2005

[Species identification of biological samples by amplifying intron 8 of the TP53 gene].

OBJECTIVE: To identify the species of biological samples by amplifying intron 8 of the TP53 gene. METHODS: Collected the bloodstains or muscle tissues of 14 kinds of common animals and human. DNA were quantified after extraction. The PCR amplification products were analyzed by PAGE and silver staining. RESULTS: The human and monkey have one amplification band of 460 bp while the eel, fish, frog, duck, rabbit, cat, mouse, cavy, pig, ox and sheep could amplify different unspecial bands. No amplification products were found in chicken and dog. CONCLUSION: The method of species identification by amplifying intron 8 of the TP53 gene is simple and sensitive.

Animals↗
PubMed1990

Scanning ion microprobe assessment of biological sample preparation techniques.

Different preparation techniques for high lateral resolution scanning ion microprobe imaging of biological samples have been investigated. The sharpest histological maps are obtained from chemically fixed and plastic embedded specimens. It is often problematic to correlate ultrastructure and bioaccumulation from analysis of frozen cut and lyophilized sections. The best compromise is to resin-embed frozen samples in order to get a perfectly flat section from tissue where the in vivo ion distribution is maintained. Use of the University of Chicago Ion Microprobe gave us the ability to observe the relative ion translocations induced during sample preparation. As an example, we show the rapid decrease of intracellular K+/Na+ ratio through a fast frozen blood droplet.

Blood Cells↗
PubMed2003

[Ethical aspects of biological sample banks].

Numerous activities in the domain of epidemiology require the constitution or the use of biological sample banks. Such biobanks raise ethical issues. A number of recommendations are applicable to this field, in France and elsewhere. Major principles applicable to biobanks include the respect of person's autonomy, the respect of human body, the respect of confidentiality. These principles are translated into practices through the following procedures: relevant information to the persons regarding their sample management prior to informed consent, opinion of an independent ethics committee, actual implementation of conditions for protecting samples and data. However, although those principles may appear quite simple and obvious, in the context of a largely international practice of research and given the large variety of biobanks, it is not always obvious for researchers to find their way. The attitudes vary between countries, there are numerous texts for various types of biobanks, the same texts raise different interpretations in different institutions, there are new ethical opinions expressed, and mainly the novelty of questions raised by the uses of samples that are possible today, especially in genetics, and were not foreseeable at the time of sampling make the field difficult in practice. This article reviews the types of biobanks, the relevant ethical issues. It also underlines the still unclear or ambiguous situations using some examples of practical situations.

Bioethical Issues↗
PubMed1981

Improved method for selenium determination in biological samples by gas chromatography.

A gas chromatographic assay employing electron-capture detection for the determination of selenium in biological samples is reported. A calibration curve of 4-nitro-o-phenylene-diamine derivative of selenium as a function of peak area was linear from 5-1000 pg. The limit of detection for the electron-capture detector was approximately 0.5 pg. Recoveries of selenium added to various biological materials ranged from 95-105%. This procedure reduces the number of transfers thereby reducing errors associated with losses or contamination. One advantage of the present method is that interfering compounds occurring in previously employed chromatographic methods are eliminated. This procedure can be used for routine microanalysis of selenium. Samples containing less than 2 ng selenium in 200 microliter of biological fluid can be routinely analyzed using this method.

Animal Feed↗
PubMed1985

High-performance liquid chromatographic separation and determination of 2-(diethylamino)ethyl diphenylpropylacetate and its metabolites from biological samples.

A high-performance liquid chromatographic method for the determination of 2-(diethylamino)ethyl diphenylpropylacetate (SKF 525-A; Proadifen), its two deethylated metabolites and a hydrolytic metabolite in biological samples has been developed. From many solvent systems investigated the compounds are best separated on an ODS/Sil-X column using an acetonitrile-phosphate buffer system containing 0.2% decylamine. The use of acetonitrile overcomes limitations associated with previously published methods for separating basic compounds where decylamine was used in a methanolic eluent. With the sampling procedures used, overall recoveries from biological tissues were around 80%. The chromatographic system also separates three tricyclic antidepressants which can be used as internal standards.

Animals↗
PubMed1996

[Neutron activation analysis of biological samples].

Multi-element analysis by neutron activation analysis (NAA) in respect to medical applications is discussed. Examples of trace element determinations in pathological human liver tissues, human serum, and rat liver are given. A very sensitive method for determining fluorinated drug in very small samples of human serum by neutron activation is also described. Each biological samples (ca. 100-200 mg, lyophilized matter) was irradiated for short time (10 s or 5 min.) and for long time (24 hrs) in Rikkyo University Research Reactor or Musashi Institute of Technology Research Reactor. Concentration of 12 elements (127I, 55Mn, 26Mn, 65Cu, 23Na, 41K, 37Cl, 27Al, 48Ca, 36S, 58Fe, 64Zn) in human pathological liver, Futraful (19F) (anti-cancer fluorinated drug) clearance of human serum, and calcium uptake (48Ca) in liver of rats were determined by NAA.

Animals↗
PubMed1992

Measurement of protein phosphatase activity in biological samples using synthetic phosphopeptides.

A method has been developed for measuring specific protein phosphatase activity in biological samples using synthetic, phospho-Kemptide and phospho-GS-peptide. This method uses ion-exchange chromatography to determine phosphatase activity by quantifying the release of [32P]phosphate directly. The method was used to measure phosphatase activity of rat kidney, adrenals, heart, and liver cytosol and the activity of purified alkaline phosphatases, protein phosphatase 1, and protein phosphatase 2A. Ion-exchange chromatography was also used for the preparation of the radiolabeled phosphopeptide substrates. This method results in high recovery and specific activity of the labeled peptides. These techniques should be useful in isolating and characterizing specific protein phosphatases found in cells.

Animals↗
PubMed1981

Extraction and purification of prostaglandins and thromboxane from biological samples for gas chromatographic analysis.

An efficient extraction procedure for the isolation of prostaglandins (PGs) from biological samples for their subsequent quantification by gas chromatography-electron capture detection (GC-ECD) is described. PGs were extracted from lung, kidney, spleen and stomach fundus into ethyl acetate at different pHs. The highest recovery and least extraction of contaminating pigments was obtained at pH 4.5. Pigments and other contaminants are removed by thin layer chromatography using a solvent system chloroform-isopropyl alcohol-ethanol-formic acid (45:5:0.5:0.3). The isolated PGs were determined by GC-ECD after appropriate derivatization. The overall recovery of PGs using this procedure is 60%.

Animals↗
PubMed1981

A method for the simultaneous determination of total carbohydrate and glycerol in biological samples with the anthrone reagent.

A method for quantitative estimation of glycerol and total carbohydrate in biological samples is described. The samples, deproteinized with cold acetone or trichloroacetic acid, were treated in the cold with 10 vols. of 0.75% (w/w) anthrone in 84% (w/w) sulphuric acid, and then heated 10 min at 100 degrees C. Absorbance at 590 nm was used for evaluation of total carbohydrate content in the sample. The absorbance at 510 nm was used for the combined carbohydrate and glycerol estimation. This latter observation leads to the determination of the glycerol content of the sample because the carbohydrate interference is known from the data obtained at 590 nm. Uses of this method to determine glycerol and carbohydrate content in lipids of chicken egg-yolk samples are presented.

Animals↗