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Rapid determination of O- and P-cresol isomers in urine from workers exposed to toluene by high-performance liquid chromatography using a graphitized carbon column.

A simple and rapid procedure is described that permits the simultaneous determination of o- and p-cresol in urine of workers of other individuals exposed to toluene. The urine samples are enzymatically hydrolysed and analysed by high-performance liquid-chromatography with a detection limit of 0.2 mg/L, one fifth of the biological threshold limit value for o-cresol (1 mg/g), a specific marker of toluene exposure. A graphitized carbon column Hybercarb-S, that exhibits an excellent selectivity for aromatic positional isomers, was used with 1% phosphoric acid-acetonitrile (70:30, v/v) as mobile phase and a detection wavelength of 271 nm. The overall accuracy for o-cresol determination was 4.5% at 5 mg/L and 8% at 0.5 mg/L. The cresol isomers were sufficiently resolved from endogenous materials to avoid the need for any extraction step, and the method appears suitable for monitoring workers accurately under the permissible level of exposure for occupational medicine purpose.

Chromatography, High Pressure Liquid

[Determination of minerals and trace elements in selenium tea from the Enschi district, People's Republic of China, and in its infusions using inductively coupled plasma-atomic emission spectrometry, graphite furnace atomic absorption spectrometry and instrumental neutron activation analysis].

The Enschi district in Hubei province, Peoples Republic of China is geochemically one of the two seleniferous regions, producing both selenium (Se) black tea and the Se green tea. Three samples of green tea with different Se contents and one non-Se tea were analysed. The following mineral and trace elements were determined by inductively coupled plasma-atomic emission spectrometry (ICP-AES): K, Ca, Mg, Na, P, S, Al, Mn, Fe, Ba, Sr, Co, Ni, Cu, Zn, Mo, and Cr. Except for Mo, Co, and Cr, all other elements in infusions of the samples analysed were also measured, since their concentrations are lying over their detection limits. The Se content in the tea samples and in the infusions were measured with graphite furnace atomic absorption spectrometry (GFAAS). The accuracy of Se determination was tested by measuring untreated tea samples with instrumental neutron activation analysis (INAA). The Se content in the measured samples was 1 to 8.5 micrograms/g. In addition to Se, 17 other elements were measured in the tea samples and 14 others in the infusions. With this data the extractable part of this elements in the infusion were calculated. Up to 10% of the Se was found in a first infusion.

China

Chromium, lead and cadmium in Danish milk products and cheese determined by Zeeman graphite furnace atomic absorption spectrometry after direct injection or pressurized ashing.

A method for the direct determination of chromium in homogeneous samples of milk and milk products is described. Minimum sample handling and prevention of contamination was given priority. After injection of the sample into the graphite furnace, the sample was ashed in a stream of oxygen at 650 degrees C and then further ashed at 1,100 degrees C with argon as the purge gas. Zeeman background correction was used in the atomisation step at 2,300 degrees C. The detection limit was 0.7 ng/g. Direct detection of chromium in milk, using only argon as purge gas, was inferior. Non-homogeneous and solid samples, e.g. yoghurt with fruit jam and cheese, were ashed under pressure with nitric acid before analysis. The same analytical principle as used for chromium was also used for the analysis of lead and cadmium in the samples. Analytical quality control was performed for both methods and the results are reported. The results for lead, cadmium and chromium in Danish milk and milk products were in the parts per billion or parts per trillion range and compare well with literature data. The intake of lead, cadmium and chromium from milk and milk products is less than 4% of the total Danish dietary intake of these elements. It is concluded that the contribution from milk and milk products to the total intake of lead and cadmium is toxicologically insignificant and that milk and milk products are only a minor source of the essential element chromium.

Animals

Serum nickel levels of diabetic patients and healthy controls by AAS with a graphite furnace.

In this study, serum nickel levels of diabetic patients and healthy controls were determined by AAS with a graphite furnace. The serum nickel concentrations were found to be 1.15 +/- 1.89 micrograms/L in healthy controls and 0.82 +/- 0.74 microgram/L in diabetics. There was, however, no statistically significant difference between the two groups (p = 0.13). The relationship of nickel levels to diabetes type and duration, diabetic complications and treatment, sex, age, and heredity was investigated. However, again no significant differences were found, nor was there any correlation between serum nickel levels and blood sugar, HBa1c, fructosamine, sialic acid levels, and age.

Adult

Physiological chromium determination in serum by Zeeman graphite furnace atomic absorption spectrometry. A serious challenge.

Several authors have already underlined chromium implication in glucose and lipids metabolism of humans. In this field, physiological chromium determination in serum could be helpful, but the discrepancies reported in numerous papers are confusing. Here we report some results obtained by Zeeman correction Graphite Furnace Atomic Absorption Spectrometry. This technique includes a dilution of serum with 12.5 mM ultrapure nitric acid and 0.25% Triton X-100 (final concentrations). Some main features can be outlined: (1) the contamination constitutes a serious drawback and (2) the sensitivity of the technique is critical (characteristic mass found: 1.76 pg/0.0044 A.s). Our results obtained from 27 healthy subjects (2.01 +/- 0.77 nmol/L) agree with most recent studies and indicate that serum chromium level does not seem to be sex-related.

Chromium

The determination of aluminum in biological fluids by means of graphite furnace atomic absorption spectrometry.

Methods are described for the determination of aluminium in serum and in diluted urine by means of graphite furnace atomic absorption spectrometry. For the analysis of serum, thermal decomposition of the sample in the furnace, using a ramp charring program, was found to be necessary in order to eliminate the organic material. The precision of the method was 10%. For urine the relative deviation was 9%. The precision was markedly improved by the use of an automatic sample injector (5.3%). Since large contamination problems make the determination of Al at the parts per billion (ppb) level very difficult, special attention was paid to the precautions to be taken in order to avoid external aluminum contamination.

Aluminum

Determination of lead in blood by graphite furnace atomic absorption spectrometry--a critique.

Graphite furnace atomic absorption spectrometry (GFAAS) is increasingly becoming the method of choice for the determination of Pb in blood. The major GFAAS methods that have been published to date include: (i) direct introduction of the sample into the furnace; (ii) dilution with water, Triton X-100 or acid; (iii) deproteinization with nitric acid; (iv) matrix modification; and (v) solvent extraction. This review focuses on the difficulties associated with each of these methods, and highlights recent attempts to overcome matrix interferences and improve the accuracy and precision of Pb determination in blood using modern furnace technology, especially the stabilized temperature platform furnace.

Humans

Graphite-furnace atomic absorption spectrometric determination of lead, cadmium, cobalt and nickel in infant formulas and evaporated milks after nitric-perchloric acid digestion and coprecipitation with ammonium pyrrolidine dithiocarbamate.

A graphite-furnace atomic absorption method, developed for lead and cadmium, was modified to enable simultaneous determination of lead, cadmium, cobalt and nickel in infant formulas and evaporated milks. The method was assessed on the basis of analytical quality assurance results during routine analysis of samples. Detection limits (ng g-1 based on 10 g sample size for ready-to-use formulas) were 0.04-0.21 for lead, 0.004-0.015 for cadmium, 0.04-0.2 for cobalt and 0.06-0.26 for nickel. Within-series repeatability and day-to-day reproducibility (among series) coefficients of variation (CVs) were, respectively, 11.7 and 16% for lead, 2.5 and 7.5% for cadmium, 7.2 and 18.8%for cobalt, and 8.1 and 8.6% for nickel at respective concentrations of 1.89, 0.40, 1.25 and 11.8 ng g-1.

Animals

Determination of nickel in urine with graphite furnace AAS using Zeeman correction.

We have developed a rapid and direct method for determining urine nickel. The urine specimen is diluted (1 + 1) with 2.0% v/v nitric acid and 0.001% v/v Triton X-100 and absorbance measurements are made with Zeeman-effect graphite furnace atomic absorption. The method is sensitive enough to be used to evaluate "normal" subjects for baseline studies or to evaluate environmental or other nonoccupational exposure to nickel. The characteristic mass (pg/0.0044A.s) is 26 pg, which is comparable to that obtained for aqueous solutions. The observed absorbance is linear up to about 100 micrograms l-1, after which the calibration curve departs from linearity. Procedures are described to rigorously exclude nickel contamination. We evaluated precision and accuracy with a U.S. National Bureau of Standards urine reference material. SRM 2670, with an informational nickel value of 70 micrograms l-1, and with a multielement water reference material, SRM 1643b, with a certified nickel value of 49 ng g-1. Within- and among-run standard deviations for SRM 2670 were calculated to be 9.0 and 2.45 micrograms l-1, respectively, and 2.1 and 1.1 micrograms l-1 for SRM 1643b. The detection limit, calculated as 3 SD of a "low" concentration urine, is about 1.1 micrograms l-1. The proposed method was applied to the determination of nickel in urine of 258 workers in a magnet manufacturing plant, and the data obtained support the usefulness of urine nickel for biological monitoring.

Environmental Exposure

A high-performance liquid chromatographic assay for reduced and oxidised glutathione in embryonic, neonatal, and adult tissues using a porous graphite electrochemical detector.

A high-performance liquid chromatographic (HPLC) assay employing a porous graphite electrochemical (EC) detector is described for the simultaneous quantification of reduced glutathione (GSH) and oxidised glutathione (GSSG) in embryonic, neonatal, and adult tissues. Samples were prepared by homogenization in 5% trichloracetic acid, centrifugation, filtration of the supernatant, and injection into the HPLC. Separation was achieved isocratically within 16 min on a 15 cm reversed-phase C18 analytical column with a particle size of 5 micron using an inexpensive mobile phase containing 2-propanol and water (2.8:100) with camphorsulfonic acid and phosphoric acid. The respective limits of detection for GSH and GSSG were 1.5 and 3 ng with a 6 microliter sample using a 3:1 signal to noise ratio. In addition to routine samples, the assay was sufficiently sensitive to detect picomole quantities of GSH and GSSG in small samples, such as a single mouse embryo, gestational day 9, weighing approximately 1 mg. The advantages and disadvantages of the method are compared with other assays for GSH and GSSG published in the literature.

Animals

Determination of lithium in rat brain regions and synaptosomes by graphite furnace atomic absorption spectrophotometry.

A graphite furnace atomic absorption spectrophotometric method for the analysis of very low concentrations of lithium in brain tissue and subcellular fractions is described. The method has a picogram sensitivity and shows a precision value of 10.1% expressed as the per cent variation coefficient for the tissue analysis of the metal. Lithium concentration in eight regions of the rat brain and regional synaptosomal lithium contents were analyzed with the method described. Results from these determinations show that lithium is heterogeneously distributed among rat brain regions 24 hr after a single s.c. lithium administration; hypothalamus, corpus striatum and midbrain were the regions with the highest lithium accumulation. Lithium is homogeneously concentrated in the synaptosomes obtained from rat brain regions. The method proposed may be considered adequate for trace lithium analysis in pharmacological studies of the metal.

Animals

An amperometric glucose sensor made by modification of a graphite electrode surface with immobilized glucose oxidase and adsorbed mediator.

A membrane-free glucose sensor was made by covalent immobilization of glucose oxidase on graphite followed by adsorption of N-methyl-phenazinium ion (PMS+). The mediator was found to be necessary for the electron transfer between the enzyme and the electrode. beta-D-glucose was determined amperometrically at an applied potential of +50 mV vs SCE. The current was independent of the rotational speed which indicates a kinetically controlled response. The response was strictly linear from the detection limit, 0.5, to 150 microM and usable up to about 2 mM beta-D-glucose. The immobilized enzyme was stable over several months but the mediator had to be renewed daily.

Electrodes

Improved high-performance liquid chromatographic procedure for the determination of lasalocid in chicken tissues and egg using polymeric and porous graphitic carbon columns.

A high-performance liquid chromatographic (HPLC) method for the determination of the ionophore coccidiostat lasalocid in poultry muscle and eggs was developed. The drug was extracted from tissue with acetonitrile. The extract was partitioned between saturated salt and carbon tetrachloride and the organic layer evaporated to dryness. Clean-up was by solid-phase extraction on a silica column. HPLC analysis was carried out on either a polymeric PLRP-S or a porous graphitic carbon Hypercarb column with a basic mobile phase and fluorescence detection with excitation at 310 nm and emission at 420-430 nm. Average recoveries from poultry muscle at the 0.002, 0.010 and 0.050 mg kg-1 levels were 65.7, 72.0 and 77.9%, respectively. Average recoveries from egg at the 0.010 and 0.100 mg kg-1 levels were 76.2 and 76.4%, respectively.

Animals

Graphite surface-assisted laser desorption/ionization time-of-flight mass spectrometry of peptides and proteins from liquid solutions.

Laser desorption time-of-flight mass spectra of peptides and proteins, as well as of lower molecular weight analytes, have been obtained by using a pulsed nitrogen UV laser (337 nm) to irradiate mixtures of 2-150 microns graphite particles and solutions of the analytes in glycerol. Protonated analytes as well as abundant alkali cation adducts were observed. Carbon cluster ions, Cn+, typically had a low abundance but dominated the mass spectrum at elevated laser powers. In spectra of a cytochrome c tryptic digest, all but one of the tryptic peptides were easily observed. Spectra of low molecular weight analytes dissolved in glycerol are very similar to FAB spectra of the same glycerol solution with added alkali salts. However, in many peptide and protein spectra, glycerol ion abundances are very low, and the alkali ions dominate the spectra at low mass. These spectra may correspond to wet and dry surface desorption conditions, respectively. The best spectra of the larger molecules were observed under dry conditions. In these initial experiments, we have obtained a sensitivity in the pico- to nanomole range and a mass resolution of about 300. The signal intensity is as good as that in conventional MALDI, and under optimal conditions, few background peaks appear, even at low mass.

Mass Spectrometry

Continuum source atomic absorption spectrometry in a graphite furnace with photodiode array detection.

A graphite furnace continuum source atomic absorption spectrometer using a photodiode array detector is described that provides high-resolution wavelength versus absorbance spectra over a 2.5-nm range for a single atomization step. The multiwavelength detection power allows the simultaneous determination of several elements, reduces problems caused by spectral interferences, and automatically corrects for nonzero background absorbance. Each spectrum is acquired in 0.33 s, and several successive spectra can be obtained during a single run. Three-dimensional wavelength-absorbance-furnace temperature spectra can be obtained by using ramped heating steps to provide a rough separation of elements in a mixture. Limits of detection calculated for 19 elements range from 0.1 pg for magnesium to 700 pg for arsenic. The sampling precision was found to be better than 10% relative standard deviation in all cases, with the precision for a single atomization being greatly increased when multiple absorption lines for a single element are observed in the spectrum. The error found for the measurement of the iron concentration in an NBS standard bronze was 8.5%, with the calculated concentration agreeing with the certified concentration within 95% confidence limits.

Metals

Direct determination of iron in urine and serum using graphite furnace atomic absorption spectrometry.

A simple, rapid and low-cost method for the routine determination of iron in urine and serum using graphite furnace atomic absorption spectrometry is described which may provide an alternative to the more widespread automated spectrophotometric methods. The urine and serum samples were simply diluted with water prior to analysis. Matrix modification was found to be redundant. The standard additions technique or the use of matrix matched standards (addition calibration) was found to be unnecessary and, therefore, the calibration was performed using aqueous standards. For serum analysis the degree of dilution could be reduced by using the less sensitive 302.0-nm resonance line, yielding more precise determinations, and for urine analysis, interferences were eliminated by means of a L'vov platform. The interferences that exist in the presence of nitric acid are also discussed. Finally, the presence of background absorption was investigated by means of Zeeman effect atomic absorption.

Humans

Direct determination of cadmium in urine using graphite furnace atomic absorption spectrometry with Zeeman-effect background correction.

A procedure is described for the direct determination of cadmium in human urine using graphite furnace atomic absorption spectrometry with Zeeman-effect background correction. Except for a straightforward 1 + 1 V/V dilution of samples with 1.5% nitric acid, no matrix modifier or sample pre-treatment was necessary, thus reducing the risk of contamination. The concentration of cadmium in urine was evaluated directly from a calibration graph prepared using a metal-spiked human urine pool. In this way the time-consuming method of standard additions was avoided, permitting an increased sample throughput (120-150 samples per day; 90 s per analysis) with minimal attention of the analyst. In routine use, the precision (both within day and day to day) and limit of detection were of the order of less than 10% and 0.05 micrograms l-1 of Cd, respectively. The method is suitable for the biological monitoring of cadmium in the general population or in occupationally exposed persons.

Cadmium

Determination of molybdenum in plasma using graphite furnace atomic absorption spectrometry.

A sensitive method is described for the determination of Mo in plasma or serum by graphite furnace atomic absorption spectrometry. The method involves extraction of the metal as the 8-hydroxyquinoline complex and is free of the interference effects that prevent the direct analysis of plasma for Mo. Recoveries of internal standards were excellent and results from the analysis of a National Institute of Standards and Technology Standard Reference Material were in good agreement with certified values. The sensitivity of the method, based on the analysis of 1 ml of plasma, is ca. 3 ng ml-1.

Animals