Characterization and quantification of silver complexes with dissolved organic matter by size exclusion chromatography coupled to ICP-MS.

The fate and behavior of silver in aquatic systems is intricately determined by its interactions with dissolved organic matter (DOM). In this study, we have introduced a method for identification and quantification of silver-DOM complexes using size exclusion chromatography-inductively coupled plasma mass spectrometry (SEC-ICP-MS). Our findings revealed that silver(I) was weakly bound to Suwannee River humic acid, fulvic acid, and natural organic matter (SRHA, SRFA, and SRNOM) in various media, resulting in facile dissociation during chromatographic separation. Suitable chromatographic conditions were determined for the elution of Ag-DOM complexes, involving the use of 0.5 mM ammonium acetate (pH 7) as the mobile phase and silver-aged column (pre-absorbing 0.1-0.7 µg silver(I)). SEC-UV and SEC-ICP-MS chromatograms revealed that Ag-binding fractions of DOM were dominated by its aromatic compounds. The quantification of silver-DOM complexes was achieved by SEC-ICP-MS combination with on-line isotope dilution. Silver at concentrations below 20 µg L-1 was mainly present in the form of organic complexes in low salinity water. These measurements aligned well with the results obtained using the equilibrium dialysis method. Species analyses of Ag-DOM complexes provide a deeper understanding of the reactivity, transport, and fate of silver in aquatic environments. ENVIRONMENTAL IMPLICATION: Ionic silver is highly toxic to aquatic organisms such as fish and zooplankton. The complexation of silver with binding sites within DOM significantly influences its speciation, mobility, and toxicity. Despite the complex and unknown structure of silver-DOM complexes, this study provided a SEC-ICP-MS method to identify and quantify these complexes in a range of media. By uncovering the formation of silver-DOM complexes across diverse media, this work enhances the comprehension of silver transformation processes and associated environmental risks in aquatic environments.

Impact of LC-MS/MS quantification methods on analytical results by example of nationwide U.S. environmental samples.

Although different quantification methods are extensively used in environmental chemistry, the impact of the choice of method on the quality and range of analytical results is understudied. This two-part study consists of (a) in-lab evaluation and (b) a traditional meta-analysis (n = 66) of commonly used quantification methods): (i) external calibration; (ii) isotope dilution method with authentic target analogs; (iii) isotope dilution with non-target standards; and (iv) standard addition prior to LC-MS/MS in liquid chromatography tandem mass spectrometry (LC-MS/MS) by example of antibiotics in sewage sludge from across the U.S. Using method (i) as the benchmark quantification method for the antibiotic erythromycin in biosolids, other quantification methods resulted in an overestimation (110-450 %) or an underestimation (10-60 %). Using the method (iv) as the benchmark for other compounds resulted in an overestimation (101-14,700 %) or an underestimation (6-98 %). Matrix effects were also observed and were dependent on the matrix and analyte type. For example, in the case of erythromycin, all sample matrices showed signal suppression. This study showed that in the absence of isotopically labeled analogs, the most accurate alternate quantification method may need to be experimentally determined depending on the analyte. Analysis of published literature on pharmaceuticals in sewage sludge indicated that isotope dilution with authentic target analog is most commonly used, followed by non-target isotope standards, standard addition, and finally external calibration.

Relationship of body mass index to percent body fat determined by deuterium isotopic dilution and impedancemetry among Tunisian schoolchildren.

Objectives: This study aims to evaluate the relation of body mass index (BMI) to fat mass among children by two techniques impedancemetry and deuterium oxide dilution (D2O). Methods: This study was carried out in 156 schoolchildren aged between 8 and 11 years. The children received interrogation specifying lifestyle and food habits. Body composition was determined using the impedancemetry and D2O technique. Results: The results showed a difference between the percentage of obese and overweight children according to BMIZ classification (30.1%), bioelectrical impedance method (14.7%) and D2O technique (42.9%). Despite the difference between the last two classifications, we found a significant correlation between body fat percentages determined by impedancemetry and D2O technique (r = 0.695, p<0.01). Bioelectrical impedance analysis underestimated %BF by 78.02% in overall children, by 70.05% in boys and by 84.73% in girls compared to D2O technique. Conclusion: This study has demonstrated that the percentage of overweight and obesity varied according the methods used. Further development of body composition methods is needed in children for the real determination of the obesity prevalence and therefore a better monitoring of this public health problem.

Antagonistic Interaction of Selenium and Cadmium in Human Hepatic Cells Through Selenoproteins.

Cadmium (Cd) is a highly toxic heavy metal for humans and animals, which is associated with acute hepatotoxicity. Selenium (Se) confers protection against Cd-induced toxicity in cells, diminishing the levels of ROS and increasing the activity of antioxidant selenoproteins such as glutathione peroxidase (GPx). The aim of this study was to evaluate the antagonistic effect of selenomethionine (SeMet) against Cd toxicity in HepG2 cells, through the modulation of selenoproteins. To this end, the cells were cultured in the presence of 100 µM SeMet and 5 µM, 15 µM, and 25 µM CdCl2 and a combination of both species for 24 h. At the end of the experiment, cell viability was determined by MTT assay. The total metal content of Cd and Se was analyzed by triple-quadrupole inductively coupled plasma-mass spectrometry (ICP-QqQ-MS). To quantify the concentration of three selenoproteins [GPx, selenoprotein P (SELENOP), and selenoalbumin (SeAlb)] and selenometabolites, an analytical methodology based on column switching and a species-unspecific isotopic dilution approach using two-dimensional size exclusion and affinity chromatography coupled to ICP-QqQ-MS was applied. The co-exposure of SeMet and Cd in HepG2 cells enhanced the cell viability and diminished the Cd accumulation in cells. Se supplementation increased the levels of selenometabolites, GPx, SELENOP, and SeAlb; however, the presence of Cd resulted in a significant diminution of selenometabolites and SELENOP. These results suggested that SeMet may affect the accumulation of Cd in cells, as well as the suppression of selenoprotein synthesis induced by Cd.

Quantitative analysis of specific androgens in serum and urine samples from male, pre, and postmenopausal subjects using LC-MRM-MS.

Androgens are endogenous hormones that play a crucial role in the paracrine and intracrine hormone system to perform and maintain vital physiological functions. Altered levels of androgens are implicated in many diseases such as sexual dysregulation, breast cancer, prostate cancer, and heart diseases etc. In this manuscript we describe a liquid chromatography-mass spectrometry (LC-MS) method using multiple reaction monitoring (MRM) for quantitatively measuring specific androgens such as dehydroepiandrosterone, testosterone, androsterone sulphate, androstenedione, and dihydrotestosterone in serum and urine samples. Serum acquired from nine different subjects (three pre-menopausal women, three postmenopausal women, and three healthy males) were used to evaluate the developed methods. In the sample preparation methods for serum either protein precipitation or liquid-liquid extraction (LLE) was used while the analysis of urinary androgens used LLE. The extracted androgens were quantitatively measured using LC-MRM-MS to which known amounts of stable isotope labeled standards were added. This manuscript also presents a LC-MRM-MS method mode for the analysis of oxime derivatized androgens potentially to enhance the sensitivity of the assay if required, from urine and venous-drawn serum samples.

Development and validation of a method for simultaneous analysis of hair underivatized amino acids and damage biomarkers, using liquid chromatography-tandem mass spectrometry.

The forensic and medical fields are seeing growing interest in the amino acid and damage biomarker composition of hair, in order to identify adulteration of drug hair testing and for diagnostic purposes. Therefore, there is an increased demand for quick and accurate analytical methods. This study presents the first liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for the simultaneous quantification of hair amino acids and four damage biomarkers, which also implements an isotopic dilution strategy to improve recovery and precision of the acid hydrolysis-sensitive analytes. The applied strategy enabled a recovery of the hydrolysis-sensitive amino acids between 83 and 120% (vs. 33-77%, without isotopic dilution) for two different protein standards, and a precision with a relative standard deviation (RSD) between 1.3 and 7.5% (vs. 9.0-29.4%, without isotopic dilution). All 21 analytes could be measured without interferences by matrix and sample components, thus demonstrating satisfactory selectivity of the method. For spiked samples of hair hydrolyzate, recovery was between 88 and 120%, whereas precision and intermediate precision were below 10.1%. The high sensitivity of the method made it possible to reduce sample preparation to a 10000-fold dilution of the raw hydrolyzate. The wide linear range displayed by the method allowed the simultaneous quantification of minor (0.3 µmol/g of hair) and major (up to 1000 µmol/g of hair) components of the biological fiber. This method was successfully applied to the analysis of real hair samples submitted to six different treatments. Statistical data analysis by means of t-test and principal component analysis (PCA) showed a clear discrimination of the treated from the untreated hair samples and of the different treatments. Since these hair treatments can interfere with hair drug testing, the method possesses the ability of identifying hair samples with potential for attempted drug test evasion. In addition, lanthionine emerged as a new biomarker for heat damaged hair.

Ecological risk assessment of trace metals in sediments under reducing conditions based on isotopically exchangeable pool.

Determination of potential mobility of toxic trace metals in sediments under changing redox condition is important in ecological risk assessment. Current methods are limited in risk prediction in such dynamic environment. In this study, we have discussed the general disagreement from widely used methods (sediment quality guideline (SQGs), potential ecological risk index (PERI), risk assessment code (RAC) using BCR fraction information). In addition, the stable isotopic dilution method (IDM) was also modified to quantify metal lability in a microcosm experiment mimicking river bank sediment turning into anaerobic. The isotopically exchangeable Cd, Cu, Pb, and Zn quantified by IDM (%E incub) was used in the RAC to reveal the trend of risk during this process. Strong risks from Cd are suggested by the PERI and RAC as a result of high toxicity and mobility of the element, while SQGs suggests medium risk for Cu, Pb, and Zn in certain samples. The disagreement between the results of RAC assessed by metal lability (%E dry) and by BCR metal fractionation reflects the effect of sediment properties and source of metal contamination. The RAC based on the non-residual fractions is likely to overestimate the potential risk for most metals even there is a significant change in sediment Eh. The RAC assessed by %E incub reveals that the variability in risk in response to the reducing Eh is not consistent. Large fluctuation in %E incub for Cd (28.5%, 49.5%), Pb (27.6%, 18.2%), and Cu (14.4%, 24.7%) can shift the risks to a higher level in certain range of Eh in two sediments. In sediment with lower contents of metal binding phases (e.g. mineral oxides, organic matters), the release of metals can be more significant, thus higher ecological risk in changing redox condition.

LC-MS-sMRM method development and validation of different classes of pain panel drugs and analysis of clinical urine samples.

Urine drug testing (UDT) is an important analytical/bio-analytical technique that has inevitably become an integral and vital part of a testing programme for diagnostic purposes. This manuscript presents a tailor-made LC-MS/MS quantitative assay method development and validation for a custom group of 33 pain panel drugs and their metabolites belonging to different classes (opiates, opioids, benzodiazepines, illicit, amphetamines, etc.) that are prescribed in pain management and depressant therapies. The LC-MS/MS method incorporates two experiments to enhance the sensitivity of the assay and has a run time of about 7 minutes with no prior purification of the samples required and a flow rate of 0.7 mL/min. The method also includes the second-stage metabolites for some drugs that belong to different classes but have first-stage similar metabolic pathways that will enable to correctly identify the right drug or to flag the drug that might be due to specimen tampering. Some real case examples and difficulties in peak picking were provided with some of the analytes in subject samples. Finally, the method was deliberated with some randomly selected de-identified clinical subject samples, and the data evaluated from "direct dilute and shoot analysis" and after "glucuronide hydrolysis" were compared. This method is now used to run routinely more than 100 clinical subject samples on a daily basis.

Levels of Alternaria Toxins in Selected Food Commodities Including Green Coffee.

Alternaria toxins are emerging mycotoxins, candidates for regulation by European Authorities. Therefore, highly sensitive, confirmatory, and reliable analytical methodologies are required for their monitoring in food. In that context, an isotope dilution LC-MS/MS method was developed for the analysis of five Alternaria toxins (Altenuene, Alternariol, Alternariol monomethylether, Tentoxin, and Tenuazonic Acid) in a broad range of commodities including cereals and cereal-based products, tomato-based products, tree nuts, vegetable oils, dried fruits, cocoa, green coffee, spices, herbs, and tea. Validation data collected in two different laboratories demonstrated the robustness of the method. Underestimation of Tenuazonic Acid level in dry samples such as cereals was reported when inappropriate extraction solvent mixtures were used as currently done in several published methodologies. An investigation survey performed on 216 food items evidenced large variations of Alternaria toxins levels, in line with data reported in the last EFSA safety assessment. The analysis of 78 green coffee samples collected from 21 producing countries demonstrated that coffee is a negligible source of exposure to Alternaria toxins. Its wide scope of application, adequate sample throughput, and high sensitivity make this method fit for purpose for the regular monitoring of Alternaria toxins in foods.

Kinetics of uranium(VI) lability and solubility in aerobic soils.

Uranium may pose a hazard to ecosystems and human health due to its chemotoxic and radiotoxic properties. The long half-life of many U isotopes and their ability to migrate raise concerns over disposal of radioactive wastes. This work examines the long-term U bioavailability in aerobic soils following direct deposition or transport to the surface and addresses two questions: (i) to what extent do soil properties control the kinetics of U speciation changes in soils and (ii) over what experimental timescales must U reaction kinetics be measured to reliably predict long-term of impact in the terrestrial environment? Soil microcosms spiked with soluble uranyl were incubated for 1.7 years. Changes in UVI fractionation were periodically monitored by soil extractions and isotopic dilution techniques, shedding light on the binding strength of uranyl onto the solid phase. Uranyl sorption was rapid and strongly buffered by soil Fe oxides, but UVI remained reversibly held and geochemically reactive. The pool of uranyl species able to replenish the soil solution through several equilibrium reactions is substantially larger than might be anticipated from typical chemical extractions and remarkably similar across different soils despite contrasting soil properties. Modelled kinetic parameters indicate that labile UVI declines very slowly, suggesting that the processes and transformations transferring uranyl to an intractable sink progress at a slow rate regardless of soil characteristics. This is of relevance in the context of radioecological assessments, given that soil solution is the key reservoir for plant uptake.

The impact of long-term biosolids application (>100 years) on soil metal dynamics.

Biosolids application to arable land is a common, and cost-effective, practice but the impact of prolonged disposal remains uncertain. We evaluated the dynamics of potentially toxic elements (PTEs) at a long-established 'dedicated' sewage treatment farm. Soil metal concentrations exceeded regulations governing application of biosolids to non-dedicated arable land. However, measurement of isotopic exchangeability of Ni, Cu, Zn, Cd and Pb demonstrated support for the 'protection hypothesis' in which biosolids constituents help immobilise potential toxic metals (PTMs). Metal concentrations in a maize crop were strongly, and almost equally, correlated with all 'capacity-based' and 'intensity-based' estimates of soil metal bioavailability. This was attributable to high correlations between soil factors controlling bioavailability (organic matter, phosphate etc.) on a site receiving a single source of PTMs. Isotopic analysis of the maize crop suggested contributions to foliar Pb from soil dust originating from neighbouring fields. There was also clear evidence of metal-specific effects of biosolids on soil metal lability. With increasing metal concentrations there was both decreasing lability of Cd and Pb, due to interaction with increasing phosphate concentrations, and increasing lability of Ni, Cu and Zn due to weaker soil binding. Such different responses to prolonged biosolids disposal to arable soil should be considered when setting regulatory limits.

Absolute quantification of selenoproteins and selenometabolites in lung cancer human serum by column switching coupled to triple quadrupole inductively coupled plasma mass spectrometry.

One of the most important causes of the high mortality rate and low life expectancy of lung cancer is the detection at advanced stages. Thus, there is an urgent need for early diagnosis and the search of new selective biomarkers. Selenium is an important constituent of selenoproteins and a powerful antioxidant able to protect against cancer. In this work, the absolute quantification of selenium in selenoproteins and the total content in selenometabolites has been performed for the first time in serum from lung cancer patients (LC) and healthy controls (HC). To this end, a method for the simultaneous speciation of selenoproteins using size exclusion chromatography (SEC) and affinity chromatography (AF) with detection by ICP-QQQ-MS, and quantification by isotopic dilution (IDA) (SEC-AF-HPLC-SUID-ICP-QQQ-MS) was developed to determine the selenium concentration in eGPx, SEPP1 and SeAlb, as well as total selenometabolites, to find alterations that may serve as biomarkers of this disease. In the same way, a method based on anion-exchange chromatography coupled to ICP-QQQ-MS was developed to quantify selenometabolites (SeCys2, SeMeSeCys, SeMet, selenite and selenate) in the same LC and HC serum samples. The results showed that the averaged concentrations of selenium in eGPx, SeAlb and selenite were significantly higher in LC patients (LC (eGPx: 21.24 ± 0.77 ng g-1; SeAlb: 49.56 ± 3.16 ng g-1 and Se(IV): 6.20 ± 1.22 ng g-1) than in HC group (eGPx: 16.96 ± 0.53 ng g-1; SeAlb: 38.33 ± 2.66 ng g-1 and Se(IV): 3.56 ± 0.55 ng g-1). In addition, the ratios between selenoproteins and selenometabolites have been calculated for the first to study their potential use as LC biomarkers. The rates eGPx/SEPP1, SEPP1/SeAlb, eGPx/Se(IV) and SEPP1/Se(IV) were significantly different between LC and HC groups.

New fully automated gas chromatographic analysis of urinary S-phenylmercapturic acid in isotopic dilution using negative chemical ionization with isobutane as reagent gas.

The determination of urinary S-phenylmercapturic acid (S-PMA) represents the most reliable biomarker to monitor the intake risk of airborne benzene. Recently, the European Chemical Agency deliberated new occupational exposure limits for benzene and recommended an S-PMA biological limit value of 2-µg/g creatinine. This limit is an order of magnitude lower than the previous one, and its determination constitutes a challenge in the analytical field. We developed and validated a method that allows the fully automated and sensitive determination of S-PMA by the use of gas-chromatography negative chemical ionization tandem mass spectrometry in isotopic dilution. For negative chemical ionization, we selected a mixture of 1% isobutane in argon as reactive gas, by studying its chemical ionization mechanism and optimal parameters compared with pure isobutane or pure methane. This gas mixture produces a more abundant signal of the target analyte than isobutane or methane, and it extended the operative lifetime of the ion source, enabling us to start a high-throughput approach of the S-PMA analysis. Moreover, energy-resolved mass spectrometry experiments were carried out to refine the MS/MS analysis conditions, testing nitrogen and argon as collision gases. The method optimization was pursued by a chemometric model by using the experimental design. The quantification limit for S-PMA was 0.10 µg/L. Accuracy (between 98.3% and 99.6%) and precision (ranging from 1.6% to 6.4%) were also evaluated. In conclusion, the newly developed assay represents a powerful tool for the robust, reliable, and sensitive quantification of urinary S-PMA, and because of its automation, it is well suited for application in large environmental and biological monitoring.

Multiple Mycotoxins Determination in Food by LC-MS/MS: An International Collaborative Study.

An intercollaborative study was organized to evaluate the performance characteristics of a liquid chromatography tandem mass spectrometry procedure for the simultaneous determination of 12 mycotoxins in food, which were ochratoxin A, aflatoxins B1, B2, G1, G2, and M1, deoxynivalenol, zearalenone, fumonisins B1 and B2, and T-2 and HT-2 toxins. The method combined the simplicity of the QuEChERS (Quick, Easy, Cheap, Efficient, Rugged and Safe) approach with the efficiency of immunoaffinity column cleanup (the step used to enhance sensitivity and sample cleanup for some matrices only). Twenty-three entities were enrolled and were European reference laboratories for mycotoxin analysis, U.S. and European service laboratories, and Nestlé laboratories. Each participant analyzed 28 incurred and/or spiked blind samples composed of spices, nuts, milk powder, dried fruits, cereals, and baby food using the protocol given. Method performances were assessed according to ISO 5725-2. Relative standard deviations of repeatability and reproducibility and trueness values for each of the 115 mycotoxin/sample combinations ranged from 5% to 23%, 7% to 26%, and 85% to 129%, respectively, in line with requirements defined in EC 401/2006. The overall set of data gathered demonstrated that the method offered a unique platform to ensure compliance with EC 1881/2006 and EC 165/2013 regulations setting maximum limits for mycotoxins in food samples, even at low regulated levels for foods intended for infants and young children. The method was applicable regardless of the food, the regulated mycotoxin, and the concentration level, and thus is an excellent candidate for future standardization.

IRMS as a tool to obtain the carbon turnover (δ13 C) in organs of weaned piglets fed glutamic acid and nucleotides.

Early weaning of piglets causes stress characterized by a decrease in feed intake followed by a decline in growth rates; thus, a fast recovery represents an essential step for proper growth of these animals. Considering that IRMS is a potential tool for non-destructive sampling and the fact that it provides time-integrated estimate of assimilated and not just ingested nutrients turned possible its application to evaluate the effects of dietary nucleotides and glutamate on carbon turnover (δ13 C) in organs of weanling piglets. At day 0, three piglets were slaughtered (prior to diet switch), the remaining eighty-four piglets weaned at 21-day-old were randomly assigned in a complete block design with a 2 × 2 factorial arrangement of treatments (two Nu levels: 0 and 0.1% and two Glu levels: 0 and 1%), being three piglets per treatment slaughtered on trial days 3, 6, 9, 14, 21, 35 and 49. The samples were analysed by IRMS and adjusted to first-order equation by a non-linear regression analysis using NLIN of SAS, in order to establish exponential graphics. After that, the turnover data were submitted to analysis of variance using GLM of SAS. The turnover value (t95% ) verified for spleen was faster (p < 0.05) when glutamate was supplemented in diets. For pancreas and liver, the turnover rates were faster (p < 0.05) for the mixture of additives. However, for renal tissue, the turnover rate (t95% ) was greater (p < 0.05) for the free additive diet. The results obtained suggest that the mixture of additives was more efficient to develop the digestive tract at post-weaning phase, taking into account the functional importance of pancreas and liver for nutrients' digestion and processing.

Gas chromatography-isotope dilution mass spectrometry method validation for target pesticides in soybeans.

The production of certified reference materials requires the application of highly accurate methods for characterisation. A gas chromatography-isotope dilution mass spectrometry method, setting ambitious performance criteria, was developed for eight selected pesticides in soybeans. Pressurised liquid extraction was followed by automated gel-permeation chromatography and solid-phase extraction clean-up. Pesticides identification respected a Commission Decision and guidelines of the Directorate General for Health and Food Safety (DG SANTE). Reliable quantification involved stable isotopically labelled analogues as internal standards. Validation, according to ISO/IEC 17,025 and DG SANTE guidelines, assessed linearity, LOD/LOQ, trueness, selectivity, precision, stability and robustness. Mean recoveries ranges (83-109%, relative standard deviations < 3%), repeatability (2.2-4.8%), day-to-day variation (0.6-4.2%) and combined uncertainty (1.2-4.2%) were fit for purpose. The method is highly accurate and suitable for certification of the selected pesticides in soybean matrix reference material. Chemical compounds studied in this article: Diazinon (PubChem CID: 3017); malathion (PubChem CID: 4004); chlorpyrifos (PubChem CID: 2730); captan (PubChem CID: 8606); endosulfan (PubChem CID: 3224); tebuconazole (PubChem CID: 86,102); iprodione (PubChem CID: 37,517); cypermethrin (PubChem CID: 2912).

[Determination of 16 phthalate acid esters in infant formula by gas chromatography-tandem mass spectrometry].

A method was established for the determination of 16 phthalate acid esters (PAEs) in infant formula by gas chromatography-tandem mass spectrometry (GC-MS/MS). PAEs in infant formula samples were homogenized by deionized water, extracted with acetonitrile, and purified by a solid-phase extraction (SPE) method using primary secondary amine (PSA) sorbents. The separation was performed on a DB-5 MS UI (30 m×0.25 mm×0.25 µm) capillary column. Effects of different elution solvents on alumina/PSA and PSA SPE columns were investigated. Fair recoveries of 16 PAEs were achieved on the PSA column upon elution by mixed n-hexane-acetone (60:40, v/v). All the 16 PAEs were quantified by matrix-matched isotopic-dilution mass spectrometry (IDMS); the 16 PAEs showed linear relationships in the concentration range of 0.01-2.0 mg/kg with linear coefficients (R2) greater than 0.9996. The limits of detection (LODs) and quantification (LOQs) were in the range of 0.15-2.5 µg/kg and 0.50-8.33 µg/kg, respectively. The recoveries of the 16 PAEs, obtained using the IDMS method, were 96.1%-104.0% with relative standard deviations (RSDs) lower than 3.3% (n=5). These results established the method described here as sensitive and precise for the simultaneous determination of 16 PAEs in infant formula, even at trace concentrations.

Extraction of polycyclic aromatic hydrocarbons from polyhydroxyalkanoates before gas chromatography/mass spectrometry analysis.

Among the organic contaminants that could pass from waste to polyhydroxyalkanoates (PHAs), there are the polycyclic aromatic hydrocarbons (PAHs). For this reason, we have developed a rapid analytical method for the determination of sixteen PAHs in PHAs. PAHs were extracted by n-hexane, after matrix dispersion and crumbling into sand; the extract was purified by solid phase extraction using florisil as adsorbent. Recoveries in the range of 89-101% were obtained for the deuterated analytes, except for the two with the lowest molecular weight. Trueness between 92% and 108% and within-laboratory precision (expressed as relative standard deviation) ≤ 18% were estimated for all the analytes. Gas chromatography/mass spectrometry was used for analyte determination. Method limits of quantification were suitable to assure that PAH presence in PHA biolpolymers is much below the limits set by European law for plastic materials. Indeed, analysis of two different PHA samples showed that contamination is limited to few compounds at non-concerning levels.

Simultaneous Speciation of Selenoproteins and Selenometabolites in Plasma and Serum.

Selenium is an essential element incorporated to different proteins with important biological functions in connection to antioxidant activity, cancer-protective properties, neurodegenerative pathologies, and prevention of effects of diabetes, among others. In addition, selenoamino acids play a basic role in the global equilibrium of key selenium-biomolecules synthesis, including selenoprotein P, selenoalbumin, and glutathione peroxidase. Homeostasis of these selenium-containing biomolecules involves different organs in living organisms including human, and bloodstream is the connection fluid in this process. Therefore, it is very important to have an analytical methodology suitable for selenium proteins and metabolites speciation in serum and plasma samples. For this purpose, a simultaneous speciation method for Se-containing biomolecules in serum/plasma is described on the basis of in series three-dimensional chromatography: size exclusion, affinity, and anion exchange high performance liquid chromatography (3D/SE-AF-AEC-HPLC), using different columns of each type and hyphenation to inductively coupled plasma-(quadrupole) mass spectrometry (ICP-MS). The method allows the quantitative simultaneous analysis of selenoprotein P (SeP), extracellular glutathione peroxidase (eGPx), selenoalbumin (SeAlb), selenite, and selenate in serum (from human and mouse) using species-unspecific isotope dilution (SUID). In addition, a simplified two-dimensional approach (2D/SE-AF-HPLC-SUID-ICP-MS) is described when selenium metabolites are globally analyzed. The method provides detection limits in the range 0.2-1.3 ng of Se g-1 and avoids typical interferences in this matrix from chloride and bromide with a chromatographic runtime less than 35 min.

Lead in Egyptian soils: Origin, reactivity and bioavailability measured by stable isotope dilution.

The current availability of Pb in Egyptian soils and associated plants were studied in 15 locations (n=159) that had been historically subjected to industrial and automobile Pb emissions. Isotopic dilution with enriched 204Pb was used to estimate the soil Pb labile pool (PbE); results showed that %PbE values were mostly <25% which is likely due to the alkaline nature of the soils. Nonetheless, lability of Pb was significantly higher in urban and industrial locations indicating greater reactivity of anthropogenic Pb in comparison to geogenic-Pb. A plot of 206Pb/207Pb vs 208Pb/207Pb showed that all soils were aligned close to a virtual binary line between two apparent end member signatures (petrol and geogenic-Pb) suggesting that they are the major sources of Pb in the Egyptian environment. Soils with greater Pb concentrations (urban and industrial locations) displayed a significantly greater ratio of labile petrol-Pb to labile geogenic-Pb in comparison to less-contaminated soils. However, this difference was marginal (±5%) suggesting that historically emitted petrol-Pb has substantially mixed with geogenic-Pb into a common pool as a result of prolonged contact with soil. The proportion of petrol-Pb in fruits and leaf vegetables was significantly (P<0.005) greater than that of the associated soils suggesting preferential uptake of the more labile petrol-Pb as opposed to the relatively immobile geogenic-Pb. However, it is also possible that the major source of Pb intake by Egyptian consumers is extraneous Pb dust enriched with petrol Pb rather than systematic Pb via roots uptake.