Determination of N-Acetyl-L-cysteine Ethyl Ester (NACET) by Sequential Injection Analysis.

New sequential injection analysis (SIA) methods with optical sensing for the determination of N-acetyl-L-cysteine ethyl ester (NACET) have been developed and optimized. NACET is a potential drug and antioxidant with advantageous pharmacokinetics. The methods involve the reduction of Cu(II) in its complexes with neocuproine (NCN), bicinchoninic acid (BCA), and bathocuproine disulfonic acid (BCS) to the corresponding chromophoric Cu(I) complexes by the analyte. The absorbance of the Cu(I) complexes with NCN, BCA, and BCS was measured at their maximum absorbance wavelengths of 458, 562, and 483 nm, respectively. The sensing manifold parameters and experimental conditions were optimized for each of the Cu(II) complexes used. Under optimal conditions, the corresponding linear calibration ranges, limits of detection, and sampling rates were 8.0 × 10-6-2.0 × 10-4 mol L-1, 5.5 × 10-6 mol L-1, and 60 h-1 for NCN; 6.0 × 10-6-1.0 × 10-4 mol L-1, 5.2 × 10-6 mol L-1, and 60 h-1 for BCA; and 4.0 × 10-6-1.0 × 10-4 mol L-1, 2.6 × 10-6 mol L-1, and 78 h-1 for BCS. The Cu(II)-BCS complex was found to be best performing in terms of sensitivity and sampling rate. Usual excipients in pharmaceutical preparations did not interfere with NACET analysis.

Hybrid organic-inorganic membranes based on sulfonated poly (ether ether ketone) matrix and iron-encapsulated carbon nanotubes and their application in CO2 separation.

The need to reduce greenhouse gas emissions dictates the search for new methods and materials. Here, a novel type of inorganic-organic hybrid materials Fe@MWCNT-OH/SPEEK (with a new type of CNT characterized by increased iron content, 5.80 wt%) for CO2 separation is presented. The introduction of nanofillers into a polymer matrix has significantly improved hybrid membrane gas transport (D, P, S, and α CO2/N2 ), and magnetic, thermal, and mechanical parameters. It was found that magnetic casting has improved the alignment and dispersion of Fe@MWCNT-OH carbon nanotubes. At the same time, CNT and polymer chemical modification enhanced interphase compatibility and membrane CO2 separation efficiency. The thermooxidative stability, and mechanical and magnetic parameters of composites were improved by increasing new CNT loading. Cherazi's model turned out to be suitable for describing the CO2 transport through analyzed hybrid membranes. The comparison of the transport and separation properties of the tested membranes with the literature data indicates their potential application in the future and the direction of further research.

Characteristics of Inorganic-Organic Hybrid Membranes Containing Carbon Nanotubes with Increased Iron-Encapsulated Content for CO2 Separation.

Novel inorganic-organic hybrid membranes Fe@MWCNT/PPO or Fe@MWCNT-OH/SPPO (with a new type of CNTs characterized by increased iron content 5.80 wt%) were synthesized for CO2 separation. The introduction of nanofillers into the polymer matrix has significantly improved the hybrid membrane's gas transport (D, P, S, and αCO2/N2), magnetic, thermal, and mechanical parameters. It was found that magnetic casting has improved the alignment and dispersion of Fe@MWCNTs. At the same time, CNTs and polymer chemical modification enhanced interphase compatibility and the membrane's CO2 separation efficiency. The thermo-oxidative stability and mechanical and magnetic parameters of composites were improved by increasing new CNTs loading. Cherazi's model turned out to be suitable for describing the CO2 transport through analyzed hybrid membranes.

Use of an ether-derived 3-hydroxy-4-pyridinone chelator as a new chromogenic reagent in the development of a microfluidic paper-based analytical device for Fe(III) determination in natural waters.

This article reports on the development and validation of a disposable microfluidic paper-based analytical device (µPAD) for on-hand, in-situ, and cheap Fe(III) determination in natural waters complying with World Health Organization guidelines. The developed µPAD used 3-hydroxy-4-pyridinone (3,4-HPO) as a colour reagent due to its considerably lower toxicity than traditionally used iron analytical reagents. It was selected among a group of hydrophilic 3,4-HPO chelators containing ether-derived chains in their structure which were prepared using green methods. The relatively high water solubility of these chelators improved the detection limit and applicability as µPAD reagents. Under optimal conditions, the µPAD is characterised by a quantification range between 0.25 and 2.0 mg/L, a detection limit of 55 µg/L and 15 min of analysis time. The signal stability extends up to 4 h and the device is stable for at least one month. The reagent consumption is below 0.2 mg per analysis and the µPAD method was validated by analysis certified reference materials and by comparison with atomic absorption results (RD < 10%). The newly developed µPAD was successfully applied to the determination of iron in river, well and tap waters with no need of any prior sample pre-treatment.

Gas-diffusion-based passive sampler for ammonia monitoring in marine waters.

A novel passive sampler based on gas-diffusion across a hydrophobic membrane is described for the determination of the time-weighted average concentration of dissolved molecular ammonia in high ionic strength aquatic environments, such as sea, coastal and estuarine waters, for a period of 3 days. The passive sampler developed is cheap, easy-to-use, reusable, and has a dynamic concentration range of 2.0-12µM, which covers the water quality guideline trigger value of 11.4µM (160µgL-1 NH3-N) for high conservation value waters, making this a powerful new tool for water quality managers involved in long-term ammonia monitoring. The gas-diffusion-based passive sampler was calibrated under laboratory conditions and deployed in a tank of seawater in the laboratory and at an estuarine site for proof of concept, and a good agreement between passive and spot sampling was achieved in both cases.

Development of a microfluidic paper-based analytical device for the determination of salivary aldehydes.

A low cost, disposable and easy to use microfluidic paper-based analytical device (µPAD) was developed for simple and non-invasive determination of total aldehydes in saliva with a potential to be used in epidemiological studies to assess oral cancer risk. The µPAD is based on the colour reaction between aldehydes (e.g. acetaldehyde, formaldehyde), 3-methyl-2-benzothiazolinone hydrazone (MBTH) and iron(III) to form an intense blue coloured formazan dye. The newly developed µPAD has a 3D design with two overlapping paper layers. The first layer comprises 15 circular detection zones (8 mm in diameter), each impregnated with 8 µL of MBTH, while the second layer contains 15 reagent zones (4 mm in diameter). Two µL of iron(III) chloride are added to each one of the second layer zones after the addition of sample to the detection zones in the first layer. All hydrophilic zones of the µPAD are defined by wax printing using a commercial wax printer. Due to the 2-step nature of the analytical reaction, the two paper layers are separated by a cellulose acetate interleaving sheet to allow for the reaction between the aldehydes in the saliva sample with MBTH to proceed first with the formation of an azine, followed by a blue coloured reaction between the azine and the oxidized by iron(III) form of MBTH, produced after the removal of the interleaving sheet. After obtaining a high resolution image of the detection side zone of the device using a flatbed scanner, the intensity of the blue colour within each detection zone is measured with Image J software. Under optimal conditions, the µPAD is characterised by a working range of 20.4-114.0 µM, limit of detection of 6.1 µM, and repeatability, expressed as RSD, of less than 12.7% (n = 5). There is no statistically significant difference at the 95% confidence level between the results obtained by the µPAD and the reference method (Student's t-test: 0.090 < 0.38). The optimized µPAD is stable for more than 41 days when stored in a freezer (≤-20 °C).

Determination of salivary cotinine through solid phase extraction using a bead-injection lab-on-valve approach hyphenated to hydrophilic interaction liquid chromatography.

Cotinine, the first metabolite of nicotine, is often used as a biomarker in the monitoring of environmental tobacco smoke (ETS) exposure due to its long half-life. This paper reports on the development of an at-line automatic micro-solid phase extraction (µSPE) method for the determination of salivary cotinine followed by its analysis via hydrophilic interaction liquid chromatography (HILIC). The SPE methodology is based on the bead injection (BI) concept in a mesofluidic lab-on-valve (LOV) flow system to automatically perform all SPE steps. Three commercially available reversed-phase sorbents were tested, namely, Oasis HLB, Lichrolut EN and Focus, and the spherically shaped sorbents (i.e., Oasis HLB and Focus) provided better packing within the SPE column and hence higher column efficiency. An HILIC column was chosen based on its potential for achieving higher sensitivity and better retention of polar compounds such as cotinine. The method uses an isocratic program with acetonitrile:100mM ammonium acetate buffer, pH 5.8 in 95:5 v/v ratio as the mobile phase at a flow rate of 1.0 mL min(-1). Using this approach, the linear calibration range was from 10 to 1000 ng which corresponded to 5-500 µg L(-1). The corresponding µSPE-BI-LOV system was proven to be reliable in the handing and analysis of viscous biological samples such as saliva, achieving a sampling rate of 6h(-1) and a limit of detection and quantification of 1.5 and 3µgL(-1), respectively.

A gas-diffusion flow injection method coupled with online solid-liquid extraction for the determination of ammonium in solid samples.

A simple, rapid and reliable gas-diffusion flow injection (GD-FI) method for ammonium determination in building materials has been developed. It is based on leaching ammonium from a ground solid sample into an alkaline solution with subsequent ammonia gas generation. Ammonia is then transported in a nitrogen stream to the GD cell of the FI system where it is absorbed into its acceptor solution containing a mixture of the acid-base indicators cresol red and thymol blue. The maximum increase in the absorbance of the acceptor solution at 580 nm is related to the ammonium concentration in the solid sample. The proposed method is characterized by a linear concentration range of 0.1-5.0 mg NH4(+) kg(-1), a limit of detection of 8 µg NH4(+) kg(-1) and a sample throughput of 10h(-1). A successful application of this method for the determination of ammonium in building materials such as concrete, cement and sand is reported.

Development of a gas-diffusion microfluidic paper-based analytical device (µPAD) for the determination of ammonia in wastewater samples.

An inexpensive, disposable and highly selective microfluidic paper-based analytical device (µPAD) is described for the determination of ammonia (molecular ammonia and ammonium cation) in wastewaters which implements for the first time a gas-diffusion separation step on a paper-based platform. Its hydrophilic reagent zones were defined by printing filter paper with a hydrophobic paper sizing agent using a conventional inkjet printer. The sample was introduced into the sodium hydroxide impregnated sample zone of the µPAD. This allowed the quantitative conversion of the ammonium ion to molecular ammonia which diffused across the hydrophobic microporous Teflon membrane of the device into an adjacent hydrophilic reagent zone containing the acid-base indicator 3-nitrophenol or bromothymol blue. The change in indicator color was measured using a desktop scanner for ammonia quantification. Under optimal conditions, the µPAD is characterized by a limit of detection of 0.8 and 1.8 mg N L(-1) and repeatability of 3.1 and 3.7% (n ≥ 10, 20 mg N L(-1)), expressed as relative standard deviation, in the case of 3-nitrophenol or bromothymol blue, respectively. This µPAD was used successfully for the determination of ammonia in sewage and soil water samples. The small dimensions, minimal reagent consumption, low cost, simplicity of operation, and possibility of using a portable scanner make the proposed µPAD suitable for on-site ammonia monitoring in contaminated environmental waters and domestic, agricultural and industrial wastewaters. The successful implementation of the gas-diffusion approach on a paper-based platform is expected to result in the development of other µPADs for volatile analytes.

Chemical characterisation and speciation of organic selenium in cultivated selenium-enriched Agaricus bisporus.

The selenium concentration in Agaricus bisporus cultivated in growth compost irrigated with sodium selenite solution increased by 28- and 43-fold compared to the control mushroom irrigated solely with water. Selenium contents of mushroom proteins increased from 13.8 to 60.1 and 14.1 to 137 µgSe/g in caps and stalks from control and selenised mushrooms, respectively. Selenocystine (SeCys; detected as [SeCys]2 dimer), selenomethionine (SeMet), and methyl-selenocysteine (MeSeCys) were separated, identified and quantified by liquid chromatography-electrospray ionisation-mass spectrometry from water solubilised and acetone precipitated proteins, and significant increases were observed for the selenised mushrooms. The maximum selenoamino acids concentration in caps and stalks of control/selenised mushrooms was 4.16/9.65 µg/g dried weight (DW) for SeCys, 0.08/0.58 µg/g DW for SeMet, and 0.031/0.10 µg/g DW for MeSeCys, respectively. The most notable result was the much higher levels of SeCys accumulated by A. bisporus compared to SeMet and MeSeCys, for both control and selenised A. bisporus.

Determination of acetaldehyde in saliva by gas-diffusion flow injection analysis.

The consumption of ethanol is known to increase the likelihood of oral cancer. In addition, there has been a growing concern about possible association between long term use of ethanol-containing mouthwashes and oral cancer. Acetaldehyde, known to be a carcinogen, is the first metabolite of ethanol and it can be produced in the oral cavity after consumption or exposure to ethanol. This paper reports on the development of a gas-diffusion flow injection method for the online determination of salivary acetaldehyde by its colour reaction with 3-methyl-2-benzothiazolinone hydrazone (MBTH) and ferric chloride. Acetaldehyde samples and standards (80 µL) were injected into the donor stream containing NaCl from which acetaldehyde diffused through the hydrophobic Teflon membrane of the gas-diffusion cell into the acceptor stream containing the two reagents mentioned above. The resultant intense green coloured dye was monitored spectrophotometrically at 600 nm. Under the optimum working conditions the method is characterized by a sampling rate of 9h(-1), a linear calibration range of 0.5-15 mg L(-1) (absorbance=5.40×10(-2) [acetaldehyde, mg L(-1)], R(2)=0.998), a relative standard deviation (RSD) of 1.90% (n=10, acetaldehyde concentration of 2.5 mg L(-1)), and a limit of detection (LOD) of 12.3 µg L(-1). The LOD and sampling rate of the proposed method are superior to those of the conventional gas chromatographic (GC) method (LOD=93.0 µg L(-1) and sampling rate=4 h(-1)). The reliability of the proposed method was illustrated by the fact that spiked with acetaldehyde saliva samples yielded excellent recoveries (96.6-101.9%), comparable to those obtained by GC (96.4-102.3%) and there was no statistically significant difference at the 95% confidence level between the two methods when non-spiked saliva samples were analysed.

Elemental and metabolite profiling of nickel hyperaccumulators from New Caledonia.

Leaf material from nine Ni hyperaccumulating species was collected in New Caledonia: Homalium kanaliense (Vieill.) Briq., Casearia silvana Schltr, Geissois hirsuta Brongn. & Gris, Hybanthus austrocaledonicus Seem, Psychotria douarrei (G. Beauvis.) Däniker, Pycnandra acuminata (Pierre ex Baill.) Swenson & Munzinger (syn Sebertia acuminata Pierre ex Baill.), Geissois pruinosa Brongn. & Gris, Homalium deplanchei (Viell) Warb. and Geissois bradfordii (H.C. Hopkins). The elemental concentration was determined by inductively-coupled plasma optical emission spectrometry (ICP-OES) and from these results it was found that the species contained Ni concentrations from to 250-28,000 mg/kg dry mass. Gas chromatography mass spectrometry (GC-MS)-based metabolite profiling was then used to analyse leaves of each species. The aim of this study was to target Ni-binding ligands through correlation analysis of the metabolite levels and leaf Ni concentration. Approximately 258 compounds were detected in each sample. As has been observed before, a correlation was found between the citric acid and Ni concentrations in the leaves for all species collected. However, the strongest Ni accumulator, P. douarrei, has been found to contain particularly high concentrations of malonic acid, suggesting an additional storage mechanism for Ni. A size exclusion chromatography separation protocol for the separation of Ni-complexes in P. acuminata sap was also applied to aqueous leaf extracts of each species. A number of metabolites were identified in complexes with Ni including Ni-malonate from P. douarrei. Furthermore, the levels for some metabolites were found to correlate with the leaf Ni concentration. These data show that Ni ions can be bound by a range of small molecules in Ni hyperaccumulation in plants.

Chelate-assisted phytoextraction of mercury in biosolids.

Mercury contaminated stockpiles of biosolids (8.4 mg kg⁻¹ Hg) from Melbourne Water's Western Treatment Plant (MW-WTP) were investigated to evaluate the possibility of their Hg chelate-assisted phytoextraction. The effects of ammonium thiosulphate (NH4)2S2O3, cysteine (Cys), nitrilotriacetic acid (NTA), and potassium iodide (KI) were studied to mobilize Hg and to increase its uptake in plant shoots. Three plant species were selected for this study, one herbaceous and two grasses: Atriplex codonocarpa, Austrodanthonia caespitosa and Vetiveria zizanioides. KI proved to be the best candidate for Hg phytostabilization in biosolids because it facilitated the concentration of this metal mainly in roots. (NH4)2S2O3 was shown to be the most effective chelating agent among those tested for Hg phytoextraction as it allowed the highest translocation of Hg into the above-ground tissues of the selected plant species. The phytoextraction conditions using A. caespitosa as the best performing plant species were optimized at an (NH4)2S2O3 concentration of 27 mmol kg⁻¹ and contact time with biosolids of seven day. Monitoring of the Hg concentration in biosolids and in leachate water during a 9-day treatment revealed that the biosolids Hg concentration decreased significantly after the first day of treatment and then it decreased only slightly with time reaching a value of 5.6 mg kg⁻¹ Hg at the end of the 9-day period. From the corresponding results obtained for the leachate water, it was suggested that a relatively large fraction of Hg (0.7 mg kg⁻¹ Hg) was promptly mobilized and consequently the plants were able to take up the metal and translocate it into shoots.

Highly sensitive gas-diffusion sequential injection analysis based on flow manipulation.

The present paper describes approaches utilizing the powerful flow manipulation capabilities of sequential injection analysis (SIA) to substantially improve the efficiency of gas-diffusion separation compared to its traditional implementation in flow injection analysis (FIA). Ammonia, ethylamine, diethylamine and triethylamine were used as model analytes in this study. Eleven flow manipulation approaches involving continuous flow, stop-flow, oscillating flow, and the introduction of air bubbles to separate the sample zone from the donor solution were tested. Improvement in sensitivity compared to traditional gas-diffusion FIA exceeding one order of magnitude was achieved. It was observed that this improvement increased with the molecular size of the analyte.

LC-MS and GC-MS metabolite profiling of nickel(II) complexes in the latex of the nickel-hyperaccumulating tree Sebertia acuminata and identification of methylated aldaric acid as a new nickel(II) ligand.

Targeted liquid chromatography-mass spectrometry (LC-MS) technology using size exclusion chromatography and metabolite profiling based on gas chromatography-mass spectrometry (GC-MS) were used to study the nickel-rich latex of the hyperaccumulating tree Sebertia acuminata. More than 120 compounds were detected, 57 of these were subsequently identified. A methylated aldaric acid (2,4,5-trihydroxy-3-methoxy-1,6-hexan-dioic acid) was identified for the first time in biological extracts and its structure was confirmed by 1D and 2D nuclear magnetic resonance (NMR) spectroscopy. After citric acid, it appears to be one of the most abundant small organic molecules present in the latex studied. Nickel(II) complexes of stoichiometry NiII:acid=1:2 were detected for these two acids as well as for malic, itaconic, erythronic, galacturonic, tartaric, aconitic and saccharic acids. These results provide further evidence that organic acids may play an important role in the transport and possibly in the storage of metal ions in hyperaccumulating plants.

Relationships of nicotianamine and other amino acids with nickel, zinc and iron in Thlaspi hyperaccumulators.

Experimental evidence suggests that nicotianamine (NA) is involved in the complexation of metal ions in some metal-hyperaccumulating plants. Closely-related nickel (Ni)- and zinc (Zn)-hyperaccumulating species were studied to determine whether a correlation exists between the Ni and Zn concentrations and NA in foliar tissues. A liquid chromatography-mass spectrometry (LC-MS) procedure was developed to quantify the NA and amino acid contents using the derivatizing agent 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. A strong correlation emerged between Ni and NA, but not between Zn and NA. Concentrations of NA and L-histidine (His) also increased in response to higher Ni concentrations in the hydroponic solution supplied to a serpentine population of Thlaspi caerulescens. An inversely proportional correlation was found between the iron (Fe) and Ni concentrations in the leaves. Correlations were also found between Zn and asparagine. The results obtained in this study suggest that NA is involved in hyperaccumulation of Ni but not Zn. The inverse proportionality between the Ni and Fe concentrations in the leaf may suggest that Ni and Fe compete for complexation to NA.

Gas-diffusion flow injection determination of Hg(II) with chemiluminescence detection.

A gas-diffusion flow injection method for the chemiluminescence detection of Hg(II) based on the luminol-H(2)O(2) reaction was developed. The analytical procedure involved the injection of Hg(II) samples and standards into a 1.50 M H(2)SO(4) carrier stream, which was subsequently merged with a reagent stream of 0.60% (w/v) SnCl(2) in 1.50 M H(2)SO(4) to reduce Hg(II) to metallic Hg. The gas-diffusion cell was thermostated at 85 degrees C to enhance the vaporisation of metallic Hg. Mercury vapour, transported across the Teflon membrane of the gas-diffusion cell into the acceptor stream containing 1.00 x 10(-4) M KMnO(4) in 0.30 M H(2)SO(4), was oxidised back to Hg(II). The acceptor stream was merged with a reagent stream containing 2.50 M H(2)O(2) in deionised water and then the combined stream was merged with another reagent stream containing 7.50 x 10(-3) M luminol in 3.00 M NaOH at a confluence point opposite to the photomultiplier tube of the detection system. The chemiluminescence intensity of the luminol-H(2)O(2) reaction was enhanced by the presence of Hg(II) in the acceptor stream. The corresponding increase was related to the original concentration of Hg(II) in the samples and standards. Under optimal conditions, the chemiluminescence gas-diffusion flow injection method was characterised by a linear calibration range between 1 microg L(-1) and 100 microg L(-1), a detection limit of 0.8 microg L(-1) and a sampling rate of 12 samples per hour. It was successfully applied to the determination of mercury in seawater and river samples.

Sensitive and ultra-fast determination of arsenic(III) by gas-diffusion flow injection analysis with chemiluminescence detection.

A novel chemiluminescence gas-diffusion flow injection system for the determination of arsenic(III) in aqueous samples is described. The analytical procedure involves injection of arsenic(III) samples and standards into a 0.3 mol L(-1) hydrochloric acid carrier stream which is merged with a reagent stream containing 0.2% (w/v) sodium borohydride and 0.015 mol L(-1) sodium hydroxide. Arsine, generated in the combined carrier/reagent donor stream, diffuses across the hydrophobic Teflon membrane of the gas-diffusion cell into an argon acceptor stream and then reacts with ozone in the flow-through chemiluminescence measuring cell of the flow system. Under optimal conditions, the method is characterized by a wide linear calibration range from 0.6 microg L(-1) to 25 mg L(-1), a detection limit of 0.6 microg L(-1) and a sample throughput of 300 samples per hour at 25 mg L(-1) and 450 samples per hour at 25 microg L(-1).

Separation of cobalt(II) from nickel(II) by solid-phase extraction into Aliquat 336 chloride immobilized in poly(vinyl chloride).

A solid-phase absorbent obtained by the immobilization of Aliquat 336 chloride in poly(vinyl chloride) is reported to extract preferentially Co(II) from its 7M hydrochloric acid solutions containing Ni(II). Under the experimental conditions there was no extraction of Ni(II) which allowed the complete separation of these two ions. Co(II) was rapidly and quantitatively back-extracted with deionised water. A mechanism for the extraction of Co(II) is proposed based on the formation of the ion-pair A(+)[HCoCl(4)](-) where A(+) is the Aliquat 336 cation. Fe(III) and Cd(II), usually present in Co(II) and Ni(II) samples, were also extracted into the solid-phase absorbent though at a slower rate than Co(II) and they did not interfere with the separation of Co(II) from Ni(II). It was also demonstrated that this approach allowed the complete separation of Ni(II) from the other metal ions mentioned above.

Metal ion ligands in hyperaccumulating plants.

Metal-hyperaccumulating plants have the ability to take up extraordinary quantities of certain metal ions without succumbing to toxic effects. Most hyperaccumulators select for particular metals but the mechanisms of selection are not understood at the molecular level. While there are many metal-binding biomolecules, this review focuses only on ligands that have been reported to play a role in sequestering, transporting or storing the accumulated metal. These include citrate, histidine and the phytosiderophores. The metal detoxification role of metallothioneins and phytochelatins in plants is also discussed.