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PubMed1984

[Low level alpha activity measurements with pulse shape discrimination--the analytical system and its characteristics].

Pulse shape discrimination of alpha and beta rays with liquid scintillation counting was investigated for the purpose of low level alpha activity measurements. Various liquid scintillators for pulse shape discrimination were examined by means of pulse rise time analysis. A new scintillator of low cost and of superior characteristics was found. The figure of merits better than 3.5 in rise time spectrum and the energy resolution better than 9% were obtained for carefully prepared samples. The background counting rate for a sample of 10 ml was reduced to 0.013 cpm/MeV in the range of alpha ray energy 5 to 7 MeV.

Alpha Particles↗
PubMed1989

Determination of 3H-labelled cytosine arabinoside and eight metabolites in cell extracts by high-performance liquid chromatography and scintillation spectrometry.

We have developed a method for the separation and quantitation of radiolabeled cytosine arabinoside and its eight metabolites in cell extracts by anion-exchange gradient high-performance liquid chromatography. Baseline separation of cytosine arabinoside and uracil arabinoside and their respective 5'-mono-, di- and triphosphates, as well as cytosine arabinoside diphosphocholine was obtained with the shortest interval between peaks being 3 min. This degree of separation was found to be essential for quantitation of 3H-labeled metabolites by scintillation counting of 1-min fractions. Application of this procedure to the quantitation of [3H] cytosine arabinoside and its metabolites from HL60 human leukemia cells is demonstrated.

Arabinofuranosyluracil↗
PubMed1987

Steroid receptors in breast cancer: sources of inter-laboratory variation in dextran-charcoal assays.

The presently recognized correlations between various clinical parameters and the concentrations of estrogen and progesterone receptors in breast cancer biopsies are largely based on receptor values obtained using the dextran-coated charcoal (DCC) method. This assay method is highly sensitive to slight changes in assay protocol, and differences in assay methodology may account for the wide variation in proportions of receptor positive patients reported by different centers. A survey of various aspects of the assay method that may lead to reproducible, systematic differences in concentrations of receptor levels is presented; and methods of compensating for or correcting these potential differences are discussed. The following aspects are considered: a) constitution of biopsy tissue, b) method of tissue homogenization, c) absorption of ligands to surfaces, d) inclusion of molybdate in the assay buffer, e) composition of the DCC slurry, and f) handling of samples for liquid scintillation counting. Differences in methods used to homogenize tissue in Europe and the U.S.A. may account for differences observed in the correlation of DCC assay results obtained using the recently-introduced monoclonal ER-EIA technique.

Adsorption↗
PubMed1989

Isotope effects on binding of NAD+ to lactate dehydrogenase.

The isotope effect on binding [4-2H]NAD+ and [4-3H]NAD+ to lactate dehydrogenase has been shown to be 1.10 +/- 0.03 by whole molecule isotope ratio mass spectrometry and 1.085 +/- 0.01 by 3H/14C scintillation counting. These values demonstrate that specific interactions of the nicotinamide ring with the enzyme make the C-H bond at C-4 less stiff in the binary complex.

Animals↗
PubMed1996

Scintillation proximity assay for human DNA topoisomerase I using recombinant biotinyl-fusion protein produced in baculovirus-infected insect cells.

DNA topoisomerases are well-established targets of important therapeutic agents which include the antibacterial quinolones and anticancer camptothecins. Screens for new classes of topoisomerase inhibitors generally employ methods, such as gel electrophoresis, which are not readily amenable to a rapid high-throughput format. We describe here a high-throughput assay to screen for inhibitors of human DNA topoisomerase I based on the scintillation proximity assay. The assay employs recombinant biotinyl-topoisomerase I fusion protein, a hybrid protein which contains a domain that is biotinylated during in vivo expression. The hybrid topoisomerase I fusion protein is found to be biotinylated, active, and nuclear-localized when produced in insect cells using a baculovirus expression system. The biotinyl-topoisomerase I fusion protein can be captured from crude nuclear extracts by immobilization on streptavidin-coated scintillation proximity assay beads. The assay detects binding of 3H-labeled DNA to the bead-immobilized enzyme by scintillation counting. The method is also able to detect stabilization of covalent protein-DNA complexes by camptothecin, an inhibitor previously shown to stabilize covalent intermediates that form during catalysis.

Animals↗
PubMed1995

Optimization and comparison of three different methods for the determination of Rn-222 in water.

Three different systems for the determination of radon in water have been examined: liquid scintillation counting (LSC), degassification followed by Lucas cell counting (LCC) and gamma counting (GC). Particular care has been devoted to the sampling methodologies of the water. Comparative results for several environmental samples are given. A critical evaluation is also given on the basis of the final aim of the measurements.

Radon↗
PubMedDate not supplied

Gross alpha determination in drinking water using a highly specific resin and LSC.

Some results of experiments concerning a new highly specific resin for the extraction of alpha-emitting nuclides from drinking water samples are presented. The product used during these experiments is a new extraction chromatographic resin which consists of a combination of several reagents and extractants supported on inert polymeric substrates, called "Resin" hereafter. It shows strong affinity for Actinides in the tri-, tetra- and hexavalent oxidation state, as well as for radium, even in presence of large amounts of calcium. Gross-alpha activities were determined using alpha-/beta-discrimination liquid scintillation counting (alpha-/beta-LSC) by direct measurement of the dried resin after extraction. Counting conditions were optimised accordingly. A method for the determination of alpha-emitting nuclides in drinking water was developed and tested using intercomparison and spiked drinking water samples.

Alpha Particles↗
PubMed1995

Microchannel array detector for quantitative electronic radioautography.

The microchannel array detector provides direct electronic detection and real time imaging of radioactive tracers in samples that traditionally have been imaged by film radioautography. The technology of the detector is described, and the main benefits of this new technology, speed and precise quantitation, are illustrated with practical results. The instrument response to a 14C dilution series (range of 10(5)) was linear over the whole range. Results from the microchannel array detector are compared to results obtained with liquid scintillation counting (LSC) and densitometry of film radioautographs. Because the instrument response is linear and consistent from day to day, it is not necessary to routinely include calibration standards as is required for film. The 14C counting efficiency was found to be essentially constant for different tissues in a whole body microtome section. As with LSC, results reported as counts per min. (CPM) are independent of counting time. Quantitative results were obtained with assays for chloramphenicol acetyl transferase in two min. and little as 0.2 DPM/mm2 of 32P was detected in seven min.

Animals↗
PubMed2006

A quantitative high-throughput trapping assay as a measurement of potential for bioactivation.

Idiosyncratic adverse drug reactions (ADRs) are one of the most common causes of pharmaceutical withdrawals and labeling changes. Most ADRs are caused by drugs that form reactive species that can bind covalently to macromolecules such as proteins. The current methodology for the measurement of covalent binding relies on the use of radiolabeled material that requires an investment in time and resources not typically expended until later in the discovery process. Efforts are also made to identify reactive intermediates by the use of chemical trapping agents, such as reduced glutathione and cyanide, to form stable adducts that are characterized by liquid chromatography-tandem mass spectrometry and/or nuclear magnetic resonance spectroscopy. Here, we describe a high-throughput assay for the measurement of reactive intermediate formation. The method involves incubation of cold compound with liver microsomes in the presence of [14C]potassium cyanide. Hard electrophilic species would react with the trapping agent, resulting in the formation of a radiolabeled conjugate. Unreacted trapping agent is removed using solid-phase extraction, and the amount of radiolabeled conjugate present is determined by liquid scintillation counting. This newly developed screen has proved to be specific, sensitive, robust, and a powerful tool for assessing bioactivation potential.

Autoanalysis↗
PubMed2006

Standardization of 18F by coincidence and LSC methods.

The nuclide 18F disintegrates to 18O by beta+ emission (96.86%) and electron capture (3.14%) with a half-life of 1.8288 h. It is widely used in nuclear medicine for positron emission tomography (PET). A radioactive solution of this nuclide has been standardized by two techniques: coincidence measurements with a pressurized proportional counter and liquid scintillation counting using the CIEMAT/NIST method. One ampoule containing a solution calibrated in activity was sent for measurement at the International Reference System maintained by the BIPM. Results are in excellent agreement with SIR values.

Calibration↗
PubMed1999

Radioactivity in bottled mineral waters.

Consumption of bottled mineral water is a growing practice and is sometimes a necessity rather than a choice. In this work, a study of the radioactive content of a wide selection of commercial bottled mineral waters for human intake was carried out. The origins of the analyzed waters were very different, coming from various locations in France, Portugal and Spain. Their total alpha and beta activity concentrations were determined and also gamma spectrometry was used to detect some radionuclides. In some cases, the waters presented high values of the total alpha and beta activity concentrations surpassing the reference levels established by the CSN, the Spanish. Regulatory Organization. In these cases, a determination of uranium and 226Ra was also performed by using low-level liquid scintillation counting. The results revealed a strong correlation between radioactive content and dry residue, and lead one to conclude that high radioactive content is mainly related to the mineralization in waters of underground origin.

Alpha Particles↗
PubMed1980

Isotope dilution technique for the estimation of l-alpha-[2-3H]-acetylmethadol and its metabolites in biological samples.

An isotope dilution technique for the determination of l-alpha-(2-3H)-acetylmethadol and its metabolites from biological samples is described. The parent drug and metabolites were extracted from biological samples with chlorobutane after the addition of unlabeled internal standards. The extracts were purified by two-dimensional thin-layer chromatography for quantification by gas-liquid chromatography and scintillation counting. Dinoracetylmethadol (NNAM) was found to be converted to 6-acetamido-4,4-diphenyl-3-heptanol (AMIDE) during the extraction procedure. The conversion of NNAM to AMIDE was confirmed by gas chromatography-mass spectrometry.

Animals↗
PubMed1986

Induction of hypoxanthine phosphoribosyltransferase deficiency in human U-937 cells.

The aim of the study was to establish enzyme-deficient mutants of the human permanent cell line U-937. Following chemical mutagenesis with the use of ethyl methanesulfonate, this cell line was chronically exposed to increasing concentrations of the toxic hypoxanthine analogue 6-thioguanine. Cells surviving hypoxanthine-aminopterin-thymidine selective media were separated by glass adherence with the use of 12-O-tetradecanoyl-phorbol-13-acetate. Three mutant clones were established, which have remained hypoxanthine phosphoribosyltransferase (HPRT) deficient for a period of 7 months, as shown by indirect measurements with the use of autoradiography and scintillation counting of cells exposed to [3H]hypoxanthine. Since the phenotypic properties and growth behavior of U-937 cells have remained unaltered after the induced mutation, a highly restricted chromosomal segment coding for HPRT seems to have been mutated.

Autoradiography↗
PubMed2003

Inhibition of nitrobenzene-induced DNA and hemoglobin adductions by dietary constituents.

Nitrobenzene (NB), a widely used industrial chemical, is a likely human carcinogen. Many dietary constituents can suppress the DNA-adduction, acting as the inhibitors of cancer. In this study, we investigated the inhibitory effects of vitamin C (VC), vitamin E (VE), tea polyphenols (TP), garlic squeeze, curcumin, and grapestone extract on NB-DNA and NB-hemoglobin (Hb) adductions in mice using an ultrasensitive method of accelerator mass spectrometry (AMS) with 14C-labelled nitrobenzene. All of these dietary constituents showed their inhibitory effects on DNA or Hb adduction. VC, VE, TP and grapestone extract could efficaciously inhibit the adductions by 33-50%, and all of these six agents could inhibit Hb adduction by 30-64%. We also investigated resveratrol, curcumin, VC and VE as inhibitors of NB-DNA adduction in vitro using liquid scintillation counting technique. These agents in the presence of NADPH and S9 components also pronouncedly blocked DNA adduction in a dose-dependent profile. Our study suggests that these seven constituents may interrupt the process of NB-induced chemical carcinogenesis.

Animals↗
PubMed2006

Standardization of tritiated water by two improved methods.

Tritiated water has been standardized in the framework of a French-Romanian cooperation by two improved methods: liquid scintillation counting based on the triple to double coincidence ratio method and the internal gas proportional counting used in conjunction with a tritium generator for chemical reduction of water. The uncertainties of measurement for both methods were smaller than 0.6% and the two results were consistent within these uncertainties, indicating that either method is equally suited for standardizing tritiated water.

Calibration↗
PubMed1998

Determination of the specific activity of sheep plasma amino acids using high-performance liquid chromatography: comparison study between liquid scintillation counter and on-line flow-through detector.

A method was developed for the determination of the specific activities of leucine and phenylalanine in plasma using a flow-through scintillation counter coupled with high-performance liquid chromatography components. Results were compared with those obtained from liquid scintillation counting. Differences in the specific activities of leucine and phenylalanine between the two methods were not statistically significant. We concluded that flow-through radioactivity detection can be used for quantitative amino acid assays. However, the minimum activity that can be detected may be prohibitively low in certain applications.

Animals↗
PubMed2006

Determination of strontium-90 in deer bones by liquid scintillation spectrometry after separation on Sr-specific ion exchange columns.

The activity concentration of (90)Sr was determined in several deer bones from Austria. Strontium specific ion exchange columns with 4',4''(5'')-di-t-butylcyclohexane-18-crown-6 from Eichrom Industries, Inc. were used for separation. The yield of the chemical procedure was quantified with AAS. Directly after column separation, the solution containing (90)Sr was mixed with the scintillation cocktail HiSafe III and measured by liquid scintillation counting. Prevention of (210)Pb contamination and reusability of the separation columns was investigated as well as the activity distribution within the bones. Results were compared with pre-Chernobyl measurements in Austria; a correlation between activity concentration of (90)Sr and site altitude was found.

Altitude↗
PubMed1990

A sensitive, radiometric assay for lysophosphatidylcholine.

To facilitate investigation of the metabolism of lysophosphatidylcholine and choline lysoplasmalogen in small quantities of tissue, a method for the quantification of these phospholipid species that is capable of accurate and reproducible analysis in samples which contain less than 1 nmol of total choline lysophospholipid was developed. The procedure employs chloroform and methanol extraction of phospholipids from isolated tissue with subsequent separation of the choline lysophospholipid fraction by high-performance liquid chromatography. The choline lysophospholipids are then acetylated with [3H]acetic anhydride and the [3H]acetyl-lysophosphatidylcholine product is isolated by thin-layer chromatography and quantified by liquid scintillation counting. The choline lysophospholipid content in the sample is determined from a standard curve constructed from samples containing a known amount of synthetic lysophosphatidylcholine with correction for recovery based on the inclusion of [14C]lysophosphatidylcholine as an internal standard.

Acetic Anhydrides↗