The role of selenium in cadmium toxicity: interactions with essential and toxic elements.

1. The study was part of a project designed to investigate if organic selenium (Se) can ameliorate the toxic effects of cadmium (Cd). The main objective of the present study was to investigate, in the chicken, the interactions between Se, Cd and the following elements: Sb, Ca, Cr, Co, Cu, Fe, Pb, Mg, Mn, Mo, Ni, V and Zn. 2. A total of 300 1-d-old chickens (broilers) were randomly distributed among 4 dietary treatments with 5 replicate pens per treatment. In T1, chickens were fed on a diet with 0·3 mg/kg added Se, without added Cd. In T2, chickens were fed on a diet with 0·3 mg/kg Se and 10 mg/kg Cd. In T3, chickens were fed on a diet with 0·3 mg/kg Se and 100 mg/kg of Cd added and in T4 treatment, chickens were fed on a diet with 3 mg/kg Se and 100 mg/kg Cd added. Se was added as Se-yeast. Cd was added as cadmium chloride (CdCl2). On d 28 and 42, two chickens per replicate pen were killed for collection of whole blood, liver, kidney and breast muscle samples. Samples were analysed by ICP-MS. The data were analysed using a multivariate linear model. 3. While low Cd concentrations in the diet led only to an increase of Cd concentration in the examined tissues, addition of high concentrations of Cd increased the concentration of Cd, Cu, Sb and V and decreased that of Se, Mn and Fe. Addition of high Se concentrations did not significantly reduce Cd concentration. 4. Prior to model application, correlations of 78 elements were noted, while after model application 39 correlations were noted. Most notably, Cd was correlated with Ca, Co, Cu and Mg, while Se was correlated with Mn. 5. The present study revealed several correlations between essential, probably essential and toxic elements illustrating the importance of the balance between pro-oxidants and antioxidants.

Tissue distribution of rare earth elements in wild, commercial and backyard rabbits.

Rare Earth Elements (REEs), La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu, Sc, and Y, & two actinides, Th and U were assessed in muscle and liver tissues of wild, backyard and commercially raised rabbits through ICP-MS. Higher concentrations were found in liver in comparison to muscle tissue. Liver of wild rabbits accumulates all studied elements beyond Tm. Backyard rabbits do not show any statistically significant accumulation while commercial accumulate all beyond La, Ce, Pr, Nd, Gd and Tb. Wild rabbits were with the highest amounts for most of these elements. The different living and rearing environments of wild, backyard and commercial rabbits may affect accumulation, fate and transfer of REEs in rabbits' tissues. A dataset for establishing reference values of REEs in Lemnos island wild rabbits' is shown and the literature gap on safety limits for REEs is discussed.

Dietary organic selenium addition and accumulation of toxic and essential trace elements in liver and meat of growing rabbits.

The effects of dietary organic selenium (Se) addition at 0.1, 0.5 and 2.5 mg/kg vs. an unsupplemented basal diet (BD) on the accumulation of some toxic and essential trace elements were studied in the liver and muscle tissues of growing rabbits. Dietary Se addition increased liver and muscle Se concentration linearly (P < .001), and decreased linearly Cd, As, Ni and Cr (P < .001) in liver, as well as As (P < .01) and Cd (P < .001) in muscle. Muscle Cu and Zn contents were significantly lower (P < .05) in rabbits fed 2.5 mg Se/kg diet compared to the other 3 groups. Selenium was negatively correlated with Cr, Ni, Cd and As (P < .01) in liver, and with Cu (P < .05) and Cd (P < .01) in muscle. In conclusion, dietary Se supplementation decreased the accumulation of toxic (Cd and As) and potentially toxic (Cr and Ni) trace elements in rabbits. However, at excessive quantities may negatively affect essential trace elements.

Game meat authentication through rare earth elements fingerprinting.

Accurate labelling of meat (e.g. wild versus farmed, geographical and genetic origin, organic versus conventional, processing treatment) is important to inform the consumers about the products they buy. Meat and meat products declared as game have higher commercial value making them target to fraudulent labelling practices and replacement with non-game meat. We have developed and validated a new method for authentication of wild rabbit meat using elemental metabolomics approach. Elemental analysis was performed using rapid ultra-trace multi-element measurement by inductively coupled plasma mass spectrometry (ICP-MS). Elemental signatures showed excellent ability to discriminate the wild rabbit from non-wild rabbit meat. Our results demonstrate the usefulness of metabolic markers -rare earth signatures, as well as other trace element signatures for game meat authentication.

Flow-injection stopped-flow kinetic spectrophotometric determination of drugs, based on micellar-catalysed reaction with 1-fluoro-2,4-dinitrobenzene.

A flow-injection stopped-flow kinetic spectrophotometric method for the determination of hydrazines, hydrazides, amines and amino-acids, based on the cetyltrimethylammonium bromide catalysed reaction with 1-fluoro-2,4-dinitrobenzene is described. With the proposed method dihydralazine, isoniazid, levodopa and aspartame can be determined at concentrations of 0.1-6 x 10(-4)M. The calibration ranges can be varied by adjusting the pH and surfactant concentration. The determination of amphetamine, cysteine, s-carboxymethylcysteine, cephalexin, tobramycin and gentamicin is also feasible. The method has been applied to the determination of levodopa, isoniazid and aspartame in commercial pharmaceutical formulations. The determination of isoniazid in formulations containing the highly coloured antibiotic rifamycin, and of aspartame in coloured beverages was also accomplished. The results were in good agreement with those obtained by reference methods and the throughput was 40 measurements per hour with 0.4-3.9% RSD.

Automated flow-injection technique for use in dissolution studies of sustained-release formulations: application to iron(II) formulations.

The application of flow-injection analysis (FIA) to automated dissolution studies of sustained-release formulations is described. The long-term stability of the dissolution-FIA analyser was checked during unattended operation for 42 h. The construction of multiple calibration curves with the so-called electronic dilution FIA procedure was used to extend the linear range of the determination. The computer-controlled FIA system and the principles of associated software are described and applied to dissolution studies of sustained-release formulations of iron(II) using its sensitive reaction with the colour reagent, ferrozine. The extended linear range of the determination is 1-130 ppm iron(II) and the precision (RSD) better than 3% (n = 3).

Selenium affects the expression of GPx4 and catalase in the liver of chicken.

A total of 128 chickens (Gallus gallus, broilers) were used to investigate the effect of organic selenium (Se) in expression of catalase (CAT) and phospholipid hydroperoxidase 4 (GPx4) genes. There were 4 replicates of 4 dietary treatments: T1 (basal diet with no added Se), T2 (T1 with 0.15 ppm Se added), T3 (T1 with 0.3 ppm Se) and T4 (T1 with 3.0 ppm Se). At 4th and 6th week, 2 chickens per replicate pen were sacrificed for whole blood and liver sample collections. Samples were analyzed for total Se by ICP-MS and gene expression by RT-PCR. Dietary supplementation with organic Se (Se-yeast) readily elevated its concentration in the tissues. GPx4 mRNA levels, pooled for both ages, of chickens fed T3 and T4 diets were significantly reduced compared to those fed diet T1 by 47% and 77% respectively, while that of T2 did not differ. Liver CAT mRNA levels at 4th week were significantly decreased as Se supplementation increased, while at 6th week, were not significantly affected by Se. The study showed that liver GPx4 mRNA levels could be down-regulated by excess of Se. It is possible that reserves built by excess of Se meet antioxidant requirements and no additional GPx4 transcription is necessary.

Construction and evaluation of an automated flow injection-stopped flow analyser for multipoint reaction rate spectrophotometric methods. Determination of ammonia nitrogen, creatinine and phosphate.

The construction and evaluation of a fully automated Flow Injection-Stopped Flow (FI-SF) spectrophotometric analyser is described. A microcomputer (Rockwell AIM 65) is used to control the analyser (sample injection, stop and start of the pump) through a suitable interface. Data acquisition is achieved using a 12 bit ADC card and a suitable subroutine in 6502 assembly language, allowing data sampling at a frequency of 7.5 kHz. The measurement interface and software were evaluated using a voltage ramp generator. A precision of 0.02-1.1% RSD (N =10) was obtained for voltage ramps in the range of 1-37 mVs(-1). The FI-SF analyser was evaluated in routine analysis by developing FI-SF kinetic spectrophotometric methods for the determination of ammonia nitrogen (20-250 ppm, 0.4-2.5% RSD) based on the Berthelot reaction, creatinine (20-220 ppm, 0.9-3.6% RSD) based on the Jaffé reaction, and phosphate (5-30 ppm, 1.0-3.3% RSD) based on the phosphomolybdenum blue reaction. The reaction rate is measured by linear fitting of multiple absorbance readings vs time. Algorithms for automated estimation of the residence time, the linear range of the reaction curve, and data treatment are presented.

Automated flow injection spectrophotometric determination of para- and meta-substituted phenols of pharmaceutical interest based on their oxidative condensation with 1-nitroso-2-naphthol.

The reaction of para- and meta-substituted phenols with 1-nitroso-2-naphthol in the presence of either CeIV or PbIV as an oxidant has been used to develop a fast automated flow injection (Fl) method. A stopped-flow kinetic study of the reaction revealed the optimum conditions for the proposed Fl method. Acetaminophen, amoxicillin, cefadroxil, isoxsuprine, nylidrin, propylparaben, tyrosine and metaraminol can be determined in the range 1 x 10(-4)-8 x 10(-4) M, with relative standard deviations of less than 2%, and a measurement throughput of 40 measurements h-1. The method was evaluated by performing interference studies of common excipients and assaying commercial formulations of acetaminophen and isoxsuprine. The results were in good agreement with those obtained by acceptable spectrophotometric or high-performance liquid chromatographic methods (mean difference 2.1%). The high sample throughput of the Fl method was exploited by performing a content uniformity test of isoxsuprine tablets.

Use of ion-selective electrodes in kinetic flow injection: determination of phenolic and hydrazino drugs with 1-fluoro-2,4-dinitrobenzene using a fluoride-selective electrode.

A flow injection (FI) kinetic potentiometric method for the determination of phenolic (acetaminophen and isoxsuprine) and hydrazino (isoniazid) drugs is described. This work shows the usefulness of ion-selective electrodes as detectors in FI systems, not only for direct ion determination but also in routine kinetic analysis. The method is based on the reaction of 1-fluoro-2,4-dinitrobenzene (FDNB) with the analytes in a weakly alkaline medium, which proceeds through the liberation of fluoride from the reagent. The slow reactions with phenols are catalysed by micelles of cetyltrimethylammonium bromide. The reaction rate is monitored with a fluoride-selective electrode in a wall-jet configuration and is used to construct a calibration graph of antilog(delta E/S)-1 versus c (where E = potential, s = slope of the electrode and c = concentration), using the fixed-time approach. The response time and the long-term stability of the electrode were found to be adequate for such kinetic determinations. The proposed method overcomes problems associated with end-point spectrophotometric methods using FDNB and allows measurements in highly coloured or turbid solutions. The optimized method has a linear concentration range of 1 x 10(-4)-50 x 10(-4) mol dm-3, a measurement throughput of 20 or 40 per hour and the precision ranges from 1.8 to 3.6% relative standard deviation (n = 3). Results obtained for commercial pharmaceutical formulations compare favourably with those given by reference methods.

Automated flow injection gradient technique for binding studies of micromolecules to proteins using potentiometric sensors: application to bovine serum albumin with anilinonaphthalenesulfonate probe and drugs.

An automated flow injection (FI) gradient technique is described for the binding study of the potentiometric probe 1-anilino-8-naphthalenesulfonate (ANS) to bovine serum albumin (BSA). Using a single-channel FI system with a mixing chamber and a flow ANS electrode, the binding parameters (binding constant and number of binding sites) were calculated using the Scatchard model. The concentration gradient was calibrated by injecting ANS in the stream, and the binding experiment was performed by injecting ANS-BSA solution in the carrier solution of equal albumin concentration. The equations describing the concentration gradient and the corresponding electrode potential curve are presented. A systematic study of the factors affecting the complexation equilibrium and the electrode response was performed. For the ANS binding to BSA, two binding classes were determined with binding constants of (2.1 +/- 0.3) x 10(5) and (3.3 +/- 0.8) x 10(3) M-1 and 3.8 +/- 0.6 and 10 +/- 2 binding sites per class, respectively, at 27 +/- 1 degrees C, in 0.10 M phosphate pH 7.4. Competitive binding experiments of sulfamethoxazole, salicylate, azapropazone, ketoprofen, and tolmetin to albumin were also performed by monitoring ANS binding inhibition (decrease of apparent binding constant). This technique takes advantage of FI gradients and direct potentiometry and utilizes the total information contained in FI peaks, providing fast and accurate binding information in a wide range of concentration ratios.

FT-Raman spectroscopy - analytical tool for routine analysis of diazinon pesticide formulations.

FT-Raman spectroscopy based on band intensity and band area measurements, was used for the quantitative determination of diazinon in pesticide formulations. Bands at 554, 604, 631, 1562 and 2971 cm(-1) were used for calibration. Spectra were acquired by averaging 100 scans at a resolution of 4 cm(-1). Calibration curves were linear (correlation coefficients, 0.992-0.9992 and 0.99-0.999 for band intensity and band area measurements, respectively) in the range of 0.2-3.5 M for 554 and 2971 cm(-1), 0.3-3.5 M for 604 cm(-1), 0.6-3.5 M for 1562 cm(-1) and 1.0-3.5 M for 631 cm(-1) bands. Normalization of calibration curves against the 802 cm(-1) cyclohexane band improved their long term stability and minimized the effect of laser beam power fluctuations. No interference was found by commonly used surfactants and the proposed method was applied to the analysis of diazinon formulations. Results obtained compare well as indicated by the t-test, with those obtained by the HPLC reference method. Precision ranged between 0.2-7.8 and 0.1-7.2% RSD, (n=4) for band intensity and band area measurements, respectively. The proposed method is rapid, simple and safe, as toxic samples are analyzed 'as received' without sample pre-treatment, permiting the routine analysis of pesticides formulations.

Construction of a l-lysine biosensor by immobilizing lysine oxidase on a gold-poly(o-phenylenediamine) electrode.

The construction of a l-lysine biosensor on a Si-gold strip electrode (SGSE) is described in this study. The construction comprises (a) the formation of poly(o-phenylenediamine, o-PD) membrane on the electrode surface via electropolymerization and (b) the immobilization of lysine oxidase on the gold/poly(o-PD) electrode with glutaraldehyde. The behavior of the gold/poly(o-PD) electrode against H(2)O(2) and lysine, as well as the repeatability of the electropolymerization and the time stability of the polymer were studied. The study showed that the electropolymerization procedure is repeatable, and that the polymer is quite stable for at least 40 days. The biosensor showed a linear calibration curve in the range 0.01-1x10(-5) M (0.1-10 muM) lysine. The interfering effect of other amino acids on the biosensor performance was also studied and amperometric selectivity coefficients were calculated. The biosensor responded mainly against tyrosine and cysteine, while the response to phenylalanine, arginine, histidine and ornithine was very low. By changing the electropolymerization conditions, the effect of interferents was further reduced.