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PubMed2004

Microsampling technique for EBSP inspection on the cross-sections of copper trench lines in ULSIs.

The crystal microstructure of copper interconnects in ultra large scale integrations (ULSIs), depends on factors such as the trench line width and patterns. Therefore, inner crystal characterisation of the desired trench lines composed of copper is required. Electron backscatter diffraction pattern (EBSP) inspection has become a powerful technique for analysing the crystal characterisation of interconnects. However, EBSP inspection is sensitive to surface conditions such as contamination and strain. In this study, we applied the focused ion beam (FIB) technology, particularly, the microsampling technique to the preparation of samples for EBSP inspection on the cross-sections of the copper trench lines. As a result, we demonstrate that a damaged layer composed of a crystalline substance is formed on the plane of the copper as revealed by the 30 kV Ga(+) FIB milling, but an EBSP signal is detectable. In conclusion, the cross-sectional sample preparation technique employing microsampling is found to be very powerful for analysing the inner crystal characteristics of the desired copper trench lines by EBSP inspection.

Copper↗

PubMed1998

Evaluation of an immunoassay (EMIT) for mycophenolic acid in plasma from renal transplant recipients compared with a high-performance liquid chromatography assay.

Mycophenolic acid is an immunosuppressant administered as a bioavailable ester, mycophenolate mofetil. The pharmacokinetics of mycophenolic acid have been reported to be variable. Accurate measurement of concentrations of this drug could be important to adjust doses. The aim of this study was to compare the enzyme-multiplied immunoassay technique (EMIT [Dade Behring; San Jose, CA, U.S.A.]) for mycophenolic acid with a high-performance liquid chromatographic (HPLC) assay using samples collected from renal transplant recipients. The HPLC assay used solid phase extraction and a C18 stationary phase with ultraviolet (UV) detection (254 nm). The immunoassay required no manual sample preparation. Plasma samples (n=102) from seven patients, collected at various times after a dose, were analyzed using both methods. Both assays fulfilled quality-control criteria. Higher concentrations were consistently measured in patient samples when using EMIT. The mean (+/-standard deviation [SD]) bias (EMIT-HPLC) was 1.88+/-0.86 mg/L. The differences in concentrations were higher in the middle of a dosage interval, suggesting that a metabolite might have been responsible for overestimation. Measurement of glucuronide concentrations by HPLC demonstrated only a weak correlation between assay differences and glucuronide concentrations. If the crossreacting substance is active, EMIT could provide a superior measure of immunosuppression; if inactive, further work is needed to improve antibody specificity. In conclusion, it was found that EMIT overestimates the concentration of mycophenolic acid in plasma samples from renal transplant recipients compared with HPLC analysis.

Chromatography, High Pressure Liquid↗

PubMed1992

Detection of influenza A and B in respiratory secretions with the polymerase chain reaction.

Influenza A and B are RNA-containing viruses that frequently infect humans. Currently, sensitive detection of these viruses requires fresh respiratory secretions and special facilities for culture. To facilitate diagnosis of influenza, the polymerase chain reaction (PCR) was used in the present studies to detect DNA produced by reverse transcription of influenzal RNA in vaccines, tissue culture fluids, and stored respiratory secretions. Primers were directed at targets on the highly conserved segment 7 (matrix gene) of influenza A (212-bp product) and B (365-bp product). The primers were completely type specific. Critical variables in the assay were the concentration of pleotropic salts used during preparation of samples, the use of carrier RNA and RNase inhibitors during sample preparation, and the use of optimum K+ and Mg2+ levels at each step. Studies of 33 patients with symptoms of viral respiratory infection whose nasal washes had been cultured and frozen for up to 1 year before assay showed that PCR provided type-specific detection of influenza with a sensitivity comparable to that of culture of the fresh secretions. The assay offers a powerful test for detection of devitalized influenza viruses and may be useful in both diagnostic work and epidemiological studies of influenza.

Base Sequence↗

PubMed1988

Why the assay of serum cystine by protein precipitation and chromatography should be abandoned.

The higher bias of serum cystine estimations by a HPLC method compared with those by ion exchange techniques is shown to be largely due to differences in the sample preparation procedures of the two techniques. The ion exchange methods utilised sulphosalicylic acid serum protein precipitation and post-column ninhydrin derivatisation of cystine, whilst the high pressure liquid chromatography technique employed automated dialysis for removal of proteins and pre-column ortho-phthalaldehyde derivatisation of cystine after its conversion to cysteine and then to S-carboxymethylcysteine. Examination of these procedures showed that whilst the high pressure liquid chromatographic method accurately estimates total serum cystine and cysteine, many factors affect the precision and accuracy of serum cystine estimations using the ion exchange techniques. In particular, serum protein precipitation techniques that are currently employed for the preparation of samples for cystine analysis by ion exchange chromatography should be abandoned.

Blood Proteins↗

PubMedDate not supplied

Direct methods for detecting picorna-like virus from dead and alive triatomine insects.

In this work we report four different destructive and non-destructive methods for detecting picorna-like virus particles in triatomines. The methods are based on direct observation under transmission electron microscope and they consist of four ways to prepare samples of presumable infected material. The samples are prepared processing dead or alive insect parts, or even dry or fresh insect feces. The methods can be used as analytical or preparative techniques, for quantifying virus infection and checking virus integrity as well. In this work the four methods are applied in order to detect Triatoma virus (TrV) particles in T. infestans colonies.

Animals↗

PubMed1991

Histamine and the stomach: chemical histamine assays.

Histamine assays in gastroduodenal tissues and body fluids are not an absolute objective of scientific interest but are related to the role of histamine in health and disease. Hence, the reliability of histamine assays has to be assessed in relation to this aim. Sensitivity and specificity of the chemical histamine assays are similar in tissues and body fluids. The modern developments in a fluorometric-fluoroenzymatic assay guarantee the highest sensitivity and specificity, especially by tests that monitor specificity in each single run of histamine determinations. Precision and accuracy of histamine measurement were especially investigated for the fluorometric assay. They included tests on the coefficient of variation over the whole concentration range, long-term precision with double-sample standard control charts, comparison of several methods for histamine assay including bioassay, and long-term accuracy with the use of Cusum charts. Finally, appropriate sample preparation, sample-taking, relevant body fluids and tissues, and the right time for sample-taking were evaluated in extended methodologic studies. Histamine assays are not just methods for a normal routine laboratory. Extended knowledge about histamine release and metabolism will be necessary to analyse data in this particular field with reasonable validity.

Animals↗

PubMed1976

Lindholm blood coagulation test values of some glow-discharge polymer surfaces.

An ultrathin layer (approximately 500A) of glow-discharge polymer that is strongly bonded to a polymer film substrate can be sued to modify the blood compatibility of the polymer surface. Glow-discharge polymers of tetrafluoroethylene, hexamethyldisiloxane, ethylene-N2, and allene-N2-H20 were deposited onto Mylar film and subjected to Lindholm whole blood coagulation tests. Lindholm tests were performed in a manner that permitted statistical analysis. This was done by repeating the preparation of samples as well as the coagulation tests a number of times. Results showed that Lindholm tests values were useful in obtaining information pertinent to compatibility of surfaces with fresh human blood and to reproducibility of sample preparation by glow-discharge polymerization. The study also revealed that glow-discharge polymerization is a promising method to impart blood compatibility without altering other bulk properties of substrate polymers.

Blood Coagulation↗

PubMedDate not supplied

Comparison of six activated partial thromboplastin time reagents: intrinsic system factors' sensitivity and responsiveness.

Six activated partial thromboplastin reagents from four manufacturers were evaluated for precision, sensitivity, and dose responsiveness to Factors VIII, IX, and XII. Normal and abnormal APTT samples were prepared. Precision was assessed using 10 replicates on three consecutive days. Factor sensitivity and responsiveness were measured using a log-log curve of percent activity versus the upper limit of normal of prepared samples. All reagents had acceptable precision (C.V. less than 5%) in the normal range, but three performed unacceptably in the abnormal range. Two of the reagents were determined to be too sensitive to certain factors, giving an abnormal APTT result when the factor activity was in the normal range. Sensitivity and dose responsiveness to factors should be considered as two separate characteristics of an APTT reagent.

Adult↗

PubMed1998

Validation of microwave digestion method for determination of trace metals in mushrooms.

A microwave digestion method for mushrooms, developed in the study, allows fast preparation of samples and reduces the contamination risk in the process of determining trace metals. Concentrations of six trace elements, Fe, Mn, Cu, Zn, Pb, and Cd were measured in 50 samples of different species of edible mushrooms (fam. Boletaceae) using atomic absorption spectrometry after microwave and dry ashing procedure. The methods were validated through certified Standard Reference Material SRM 1577b (Bovine Liver) which was treated and analysed using the same procedures as for the mushrooms. The samples were either digested with concentrated HNO3 in closed Teflon PFA vessels in a microwave oven, or ashed in quartz crucibles at 450 degrees C. The respective recoveries of Fe, Mn, Zn, Cu, Pb, and Cd obtained by measuring SRM were 112, 107, 104, 115, 111 and 95% after microwave digestion procedure and 86, 101, 109, 111, 98, and 110% after dry ashing procedure. The correlation between concentrations obtained by the two different methods of sample preparation was high for all metals.

Agaricales↗

PubMedDate not supplied

Determination of amoxicillin residues in animal tissues by solid-phase extraction and liquid chromatography with fluorescence detection.

Trace levels of amoxicillin residues were determined in animal tissues by liquid chromatography (LC) with fluorescence detection. An improved solid-phase extraction (SPE) procedure requiring less flammable solvent (diethyl ether) was developed for sample preparation. Muscle samples of beef, pork, chicken, and tilapia were extracted with a phosphate buffer followed by the modified SPE procedure for cleanup and concentration prior to the LC-fluorescence analysis. Average recoveries of fortified amoxicillin at 5, 10, and 20 micrograms/kg ranged from 83.9 to 85.8% in beef, 86.1 to 88.1% in pork, 81.7 to 82.9% in chicken, and 92.5 to 95.4% in tilapia. Relative standard deviations were < 4%.

Amoxicillin↗

PubMed2000

Branched-chain keto-acids and pyruvate in blood: measurement by HPLC with fluorimetric detection and changes in older subjects.

BACKGROUND: Measurement of keto-acids is important in various clinical situations. The aim of the present work was to develop a rapid HPLC method for the determination of keto-acids in human serum and to assess the concentrations of these acids in young adults and institutionalized elderly adults. This method was applied to the determination of blood keto-acid concentrations of young adults and institutionalized elderly people, divided into age groups METHODS: Four keto-acids (alpha-ketoisocaproate, alpha-ketoisovalerate, alpha-keto-beta-methylvalerate, and pyruvate) were derivatized with o-phenylenediamine to give fluorescent derivatives. After the sample preparation step (75 min to prepare 20 samples), the derivatives were separated chromatographically on a reversed-phase column using a binary gradient. RESULTS: The fluorometric detection of the four keto-acids was rapid, <12 min. The method is repeatable and reproducible: the CVs were <6% and <11%, respectively, for each of the keto-acids. We found no significant difference between males and females. Concentrations of the branched-chain keto-acids decreased after age 60 years, especially alpha-ketoisocaproate, which decreased approximately 40%. CONCLUSIONS: The proposed method allows rapid and reliable measurement of keto-acids. The data demonstrate that changes in branched-chain keto-acids concentrations in serum occur with age.

Adult↗

PubMed2001

Airborne polyvinyl alcohol (PVA) and cellulose fibre levels in fibre-cement factories in seven European countries.

Because of their relatively high diameter, polyvinyl alcohol (PVA) fibres, as used in fibre-cement, are not fibres as defined by WHO (or other) regulations. Nevertheless, as with all particulate raw materials, it can be questioned if and to what extent particles with critical fibrous dimensions might be generated by the handling or machining of this material. In order to investigate any tendency of PVA fibres to release airborne particles with critical fibrous dimensions (WHO fibres), static and/or personal samples were taken in eight fibre-cement factories at locations where potential exposures to PVA fibres were expected to be the highest. The following locations were surveyed: the PVA fibre weighing station, where PVA bales are opened mechanically and the PVA fibres are dispersed and weighed in a dry state; the fibre-cement slate punching machine; the slate 'riven edge' cutting machine or sheet sawing machine, whichever was present in the respective factories. Since cellulose fibres are an important constituent of fibre-cement, the organic fibre concentrations observed at the machining operations include cellulose. At each factory a control sample was taken in open air. Sampling, sample preparation and sample analysis by scanning electron microscopy (SEM) were performed according to standard German procedures. Only very low number concentrations of organic WHO fibres, ranging from below detection limit to 0.006 f/ml, were found. These levels are lower than the typical levels of organic fibres commonly found in the normal personal environment (0.009-0.02 f/ml), stemming from the release of particles by a person's activities and from clothing and other textiles (bed sheets, blankets, pillow,.). We conclude that the handling of PVA fibres as well as the machining of PVA and cellulose fibre containing cement products in the fibre-cement factories surveyed have a low potential to release fibres with critical fibrous (WHO) dimensions.

Air Pollutants↗

PubMed2002

Psychotropic drugs in developmental mushrooms: a case study review.

Psilocyn and psilocybin can be identified in different stages of developing psilocybe mushrooms. Knowing the various stages of the mushroom development can be useful when receiving evidence from illicit mushroom growing operations. Exhibits from three separate cases were submitted to the drug analysis section of the Minnesota Bureau of Criminal Apprehension Forensic Science Laboratory. Each case contained different stages of developing mushrooms. This report describes the evidence in each case, the sample preparation, the sample analysis and the final report that was written.

Agaricales↗

PubMed2002

[Modified flow cytometry assay for CD62p expression of preserved platelets].

CD62p expression was an important monitoring parameter for preserved platelets quality. To setup an optimized flow cytometry assay for preserved platelets based on the CD62p expression on platelets, the platelet samples were collected, 0.1 mmol/L persantine and 1.1 mmol/L EDTA were added into the modified TB S used to replace PBS dilution; the methodological evaluation were carried out. Results showed that 0.1 mmol/L persantine and 1.1 mmol/L EDTA achieved to prevent platelets activation during the test procedure. The favorable negative or positive samples were prepared for check of fluorescence antibody's quality to ensure the validity of results. CD61 was used to identify platelets for assay and improve veracity of assay. The special injector was also replaced by special big syringe needle for blood collection to reduce in vitro artifacts, and the prepared sample can be steady-going for 48 hours at 4 degrees C after fixed by 1% paraformaldehyde. It is concluded that this flow cytometry assay for CD62p positive platelets is simple and efficient.

Blood Platelets↗

PubMed1988

The apparent target size of rat brain benzodiazepine receptor, acetylcholinesterase, and pyruvate kinase is highly influenced by experimental conditions.

Radiation inactivation is a method to determine the apparent target size of molecules. In this report we examined whether radiation inactivation of various enzymes and brain receptors is influenced by the preparation of samples preceding irradiation. The apparent target sizes of endogenous acetylcholinesterase and pyruvate kinase from rat brain and from rabbit muscle and benzodiazepine receptor from rat brain were investigated in some detail. In addition the target sizes of alcohol dehydrogenase (from yeast and horse liver), beta-galactosidase (from Escherichia coli), lactate dehydrogenase (endogenous from rat brain), and 5-HT2 receptors, acetylcholine muscarine receptors, and [35S] butyl bicyclophosphorothionate tertiary binding sites from rat brain were determined. The results show that apparent target sizes are highly influenced by the procedure applied for sample preparation before irradiation. The data indicate that irradiation of frozen whole tissue as opposed to lyophilized tissue or frozen tissue homogenates will estimate the smallest and most relevant functional target size of a receptor or an enzyme.

Acetylcholinesterase↗

PubMed1979

Radioimmunoassay of hair for determining opiate-abuse histories.

Heroin and morphine metabolites can be detected in hair with the use of commercially available radioimmunoassay reagents and with minor sample preparation. Hair samples obtained from morphine-treated mice and heroin users contained nanogram levels of the drug per milligram of hair (single human hair). The results of the hair analyses for all subjects admitting the use of heroin were positive, whereas the results of only 30% of thin-layer chromatographic urinanalyses of these same subjects were positive. In addition, differences in drug concentration for sections of hair near the scalp and near the distal end correlated with the length of time the drug had been used. These results exemplify the potential advantages of the use of hair analysis over urine and serum analyses in terms of accessibility, sample stability, and long-term retention of information.

Animals↗

PubMedDate not supplied

Urinary cytology: device capabilities and requirements.

The practical and theoretical considerations leading to the automated diagnostic assessment of cells in the urinary sediment by a computer-based, high-resolution system are discussed. Problems of sample preparations, sample-size requirements, coefficients of variation and computer-generated features are summarized. There are no major theoretical obstacles to achieving a workable system for the diagnosis of high-grade urothelial cancer by the proposed approach.

Computers↗

PubMed1995

Development of a HPLC-system for quantitative measurement of lidocaine and bupivacaine in patients plasma during postoperative epidural pain therapy.

A HPLC method was developed for the simultaneous quantitative analysis of lidocaine and bupivacaine in plasma, with bupivacaine serving as the internal standard for the assessment of lidocaine and vice versa. The samples are prepared by diethyl ether-extraction of the alkalified plasma and re-extraction using diluted sulphuric acid. This allows the elimination of interfering medication and plasma proteins. The prepared samples are chromatographed with a Merck LiChroCART Superspher 60 RP-select B cartridge column, the local anesthetics are detected using UV-photometry and the concentration is calculated by comparing the peak areas of the analyzed substance and the internal standard. Using a sample volume of 1 ml plasma, concentrations of approximately 2.5 micrograms/ml and 1 microgram/ml can be analyzed with a 95%-confidence interval of 2.5% or 5%, respectively. At higher or lower concentrations, accurate results can be obtained using smaller or larger plasma samples. The evolved analytical method allows the rapid and simple determination of lidocaine and bupivacaine plasma levels at a wide range of concentrations. It is suitable for research purposes as well as for routine analyses.

Analgesia, Epidural↗