Extraction of arsenic compounds from lichens

Different extraction procedures were applied to improve the extraction efficiency of arsenic compounds from lichens. Two lichen species were chosen from an arsenic-contaminated environment: epiphytic Hypogymnia physodes (L.) Nyl. and terricolous Cladonia rei Schaer. Samples were extracted with water at temperatures of 20, 60 and 90 °C, using mixtures of methanol/water (9:1, 1:1 and 1:9), Tris buffer and acetone and the extracts speciated. Water and Tris buffer showed the best extraction efficiency of all extractants used; however, the extraction efficiency was still less than 23%. Since a major fraction of arsenic appeared to be associated with trapped soil particles, a sequential extraction procedure originally designed for soils (extraction steps: (1) 0.05 mol l⁻¹ (NH₄)₂SO₄; (2) 0.05 mol l⁻¹ (NH)₄H₂PO₄; (3) 0.2 mol l⁻¹ NH₄-oxalate buffer, pH 3.25; (4) mixture of 0.2 mol l⁻¹ NH₄-oxalate buffer and 0.1 mol l⁻¹ ascorbic acid, pH 3.25; (5) 0.5 mol l⁻¹ KOH) was applied and found to remove 45% of the total arsenic from H. physodes and 83% from C. rei. The lipid-soluble fraction of arsenic was estimated by k₀-INAA analysis of diethylether extracts and was found to be negligible. An HPLC-UV-HGAFS system was used to determine the arsenic compounds extracted. In both lichen species, arsenous acid, arsenic acid, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, trimethylarsine oxide and glycerol-ribose were detected. In addition, phosphate-ribose was found in H. physodes.     

Enzyme-assisted extraction and liquid chromatography mass spectrometry for the determination of arsenic species in chicken meat

Chicken is the most consumed meat in North America. Concentrations of arsenic in chicken range from μg kg⁻¹ to mg kg⁻¹. However, little is known about the speciation of arsenic in chicken meat. The objective of this research was to develop a method enabling determination of arsenic species in chicken breast muscle. We report here enzyme-enhanced extraction of arsenic species from chicken meat, separation using anion exchange chromatography (HPLC), and simultaneous detection with both inductively coupled plasma mass spectrometry (ICPMS) and electrospray ionization tandem mass spectrometry (ESIMS). We compared the extraction of arsenic species using several proteolytic enzymes: bromelain, papain, pepsin, proteinase K, and trypsin. With the use of papain-assisted extraction, 10 arsenic species were extracted and detected, as compared to 8 detectable arsenic species in the water/methanol extract. The overall extraction efficiency was also improved using a combination of ultrasonication and papain digestion, as compared to the conventional water/methanol extraction. Detection limits were in the range of 1.0–1.8 μg arsenic per kg chicken breast meat (dry weight) for seven arsenic species: arsenobetaine (AsB), inorganic arsenite (As^III), dimethylarsinic acid (DMA), monomethylarsonic acid (MMA), inorganic arsenate (As^V), 3-nitro-4-hydroxyphenylarsonic acid (Roxarsone), and N-acetyl-4-hydroxy-m-arsanilic acid (NAHAA). Analysis of breast meat samples from six chickens receiving feed containing Roxarsone showed the presence of (mean ± standard deviation μg kg⁻¹) AsB (107 ± 4), As^III (113 ± 7), As^V (7 ± 2), MMA (51 ± 5), DMA (64 ± 6), Roxarsone (18 ± 1), and four unidentified arsenic species (approximate concentration 1–10 μg kg⁻¹).     

A microwave-assisted sequential extraction of water and dilute acid soluble arsenic species from marine plant and animal tissues

This paper describes the use of dilute nitric acid for the extraction and quantification of arsenic species. A number of extractants (e.g. water, 1.5 M orthophosphoric acid, methanol-water and dilute nitric acid) were tested for the extraction of arsenic from marine biological samples, such as plants that have proved difficult to quantitatively extract. Dilute 2% (v/v) nitric acid was found to give the highest recoveries of arsenic overall and was chosen for further optimisation. The optimal extraction conditions for arsenic were 2% (v/v) HNO₃, 6 min⁻¹, 90 °C. Arsenic species were found to be stable under the optimised conditions with the exception of the arsenoriboses which degraded to a product eluting at the same retention time as glycerol arsenoribose. Good agreement was found between the 2% (v/v) HNO₃ extraction and the methanol-water extraction for the certified reference material DORM-2 (AB 17.1 and 16.2 μg g⁻¹, respectively, and TETRA 0.27 and 0.25 μg g⁻¹, respectively), which were in close agreement with the certified concentrations of AB 16.4 ± 1.1 μg g⁻¹ and TETRA 0.248 ± 0.054 μg g⁻¹.To preserve the integrity of arsenic species, a sequential extraction technique was developed where the previously methanol-water extracted pellet was further extracted with 2% (v/v) HNO₃ under the optimised conditions. Increases in arsenic recoveries between 13% and 36% were found and speciation of this faction revealed that only inorganic and simple methylated species were extracted.     

Studying the distribution pattern of selenium in nut proteins with information obtained from SEC-UV-ICP-MS and CE-ICP-MS

In this work, size exclusion chromatography (SEC) with UV and inductively coupled plasma mass spectrometry (ICP-MS) detection was used to study the association of selenium to proteins present in Brazil nuts (Bertholletia excelsa) under five different extraction conditions. As expected, better solubilization of proteins was observed using 0.05 mol L⁻¹ sodium hydroxide and 1% sodium dodecylsulfate (SDS) in Tris/HCl buffer (0.05 mol L⁻¹, pH 8) as compared to 0.05 mol L⁻¹ HCl, 0.05 mol L⁻¹ Tris/HCl or hot water (60 °C). Due to non-destructive character of Tris-SDS treatment, this was applied for studying molecular weight (MW) distribution patterns of selenium-containing nut proteins. Three different SEC columns were used for obtaining complete MW distribution of selenium: Superdex 75, Superdex Peptide, and Superdex 200 were tested with 50 mmol L⁻¹ Tris buffer (pH 8), 150 mmol L⁻¹ ammonium bicarbonate buffer (pH 7.8), phosphate (pH 7.5), and CAPS (pH 10.0) mobile phases. Using Superdex 200 column, the elution of at least three MW fractions was observed with UV detection (200-10 kDa) and ICP-MS chromatogram showed the co-elution of selenium with the two earlier fractions. The apparent MWs of these selenium-containing fractions were respectively about 107 and 50 kDa, as evaluated from the column calibration. For further characterization of individual selenium species, the defatted nuts were hydrolyzed with proteinase K and analyzed by capillary electrophoresis (CE) with ICP-MS detection. The suitability of CE for the separation of selenite, selenate, selenocystine and selenomethionine in the presence of the nut sample matrix is demonstrated. Complete separation of the above mentioned selenium species was obtained within a migration time of 7 min. In the analysis of nut extracts with CE-ICP-MS, selenium was found to be present mainly as selenomethionine.     

Development, validation and application of a method based on DI-SPME and GC-MS for determination of pesticides of different chemical groups in surface and groundwater samples

A simple and rapid method based on solid-phase micro extraction (SPME) technique followed by gas chromatography-mass spectrometry with selected ion monitoring (GC-MS, SIM) was developed by the simultaneous determination of 16 pesticides of seven different chemical groups [Six organophosphorus (trichlorfon, diazinon, methyl parathion, malathion, fenthion and ethyon), three pyrethroids (bifenhin, permethrin, cypermethrin), two imidazoles (imazalil and prochloraz), two strobilurins (azoxystrobin and pyraclostrobin), one carbamate (carbofuran), one tetrazine (clofentezine), and one triazole (difenoconazole)] in water. The pesticides extraction was done with direct immersion mode (DI-SPME) of the polyacrilate fiber (PA 85 µm). The extraction temperature was adjusted to 50 °C during 30 min, while stirring at 250 rpm was applied. After extraction, the fiber was introduced in the GC injector for thermal desorption for 5 min. at 280 °C. The method was validated using ultra pure water samples fortified with pesticides at different concentration levels and shows good linearity in the concentrations between 0.05 and 250.00 ng mL^− 1. The LOD and LOQ ranged, from 0.02 to 0.30 ng mL^− 1 and 0.05 to 1.00 ng mL^− 1, respectively. Intra-day and inter-day precisions were determined in two concentration levels (5.00 and 50.00 ng mL^− 1). Intra-day relative standard deviation (%R.S.D.) ranged between 3.6 and 13.6%, and inter-day (%R.S.D.) ranged between 6.3 and 18.5%. Relative recovery tests were carried out spiking the ultra pure sample with standards in three different concentration levels 0.20, 5.00 and 50.00 ng mL^− 1. The recovery at 0.20 ng mL^− 1 level varied from 86.4 ± 9.4% to 108.5 ± 10.5%, at 5.00 ng mL^− 1 level varied from 77.5 ± 10.8% to 104.6 ± 9.6% and at 50.00 ng mL^− 1 level varied from 70.2 ± 4.6% to 98.4 ± 8.5%. The proposed SPME method was applied in twenty-six water samples collected in the “Platô de Neópolis”, State of Sergipe, Brazil. Methyl parathion was detected in five samples with an average concentration of 0.17 ng mL^− 1 and bifenthrin, pyraclostrobin and azoxystrobin residues were found in three samples with average concentrations of 2.28, 3.12 and 0.15 ng mL^− 1, respectively.     

Determination of total arsenic and toxicologically relevant arsenic species in fish by using electrothermal and hydride generation atomic absorption spectrometry

A scheme for the determination of total As by electrothermal atomic absorption spectrometry (ETAAS) and the sum of toxicologically relevant arsenic species (As(III), As(V), monomethylarsonate (MMA) and dimethylarsinate (DMA) using hydride generation AAS (HGAAS) in fish samples was developed. Simple and fast microwave assisted extraction in tetramethylammonium hydroxide (TMAH, 0.075% m / v) or in water-methanol mixture (80 + 20 v / v) for 20 min is proposed for quantitative leaching of arsenic species from fish tissue. Total As was measured by ETAAS directly in the TMAH extract under optimal instrumental parameters (pyrolysis temperature 1400 °C and atomization temperature 2000 °C) with Pd as modifier ensuring thermal stabilization and isoformation of all extracted arsenic species. The analytical features of the method are as follows: limit of detection (LOD) 0.45 μg g^− 1 (dry wt.), within-run and between-run precision in the range 4-8% and 5-12%, respectively, for arsenic contents 0.5-30 μg g^− 1 and recoveries 98-102%. The sum of toxicologically relevant arsenic species (As(III) + As(V) + MMA + DMA) was determined by flow injection HGAAS directly from the TMAH extract or water-methanol mixture and trapping of arsines onto Zr-Ir coated graphite tube followed by ETAAS measurement. l-cysteine is used as reagent for leveling off responses of different arsenic species in the presence of TMAH or water-methanol mixture. The LODs achieved are 0.0038 and 0.0031 μg g^− 1 (dry wt.), respectively, for fish extracts in TMAH and in water-methanol mixture. Within-batch and between-batch RSDs are in the range 3-5% and 4-7% for arsenic contents of 0.009-0.25 μg g^− 1 (dry wt.) for TMAH extracts and 2-4% and 3-6% for methanol water extracts, respectively. Selective reaction media for generation of respective hydrides from arsenic species were recommended for further speciation purposes in methanol-water extracts, viz. citrate buffer (pH 5.2) for the determination of As(III), 0.2 mol L^− 1 acetic acid for the determination of As(III) + DMA and 7 mol L^− 1 hydrochloric acid for the determination of inorganic As(III) + As(V). LODs are 0.0035, 0.0051 and 0.0046 μg g^− 1 (dry wt.) for As(III), DMA and As(V). The relative standard deviation is 4-8% for three arsenic species at As levels of 0.009-0.5 μg g^− 1 (dry wt.). The accuracy of the proposed speciation scheme is confirmed by the analysis of certified reference materials.     

Arsenic speciation in edible alga samples by microwave-assisted extraction and high performance liquid chromatography coupled to atomic fluorescence spectrometry

Twelve commercially available edible marine algae from France, Japan and Spain and the certified reference material (CRM) NIES No. 9 Sargassum fulvellum were analyzed for total arsenic and arsenic species. Total arsenic concentrations were determined by inductively coupled plasma atomic emission spectrometry (ICP-AES) after microwave digestion and ranged from 23 to 126 μg g⁻¹. Arsenic species in alga samples were extracted with deionized water by microwave-assisted extraction and showed extraction efficiencies from 49 to 98%, in terms of total arsenic. The presence of eleven arsenic species was studied by high performance liquid chromatography–ultraviolet photo-oxidation–hydride generation atomic–fluorescence spectrometry (HPLC–(UV)–HG–AFS) developed methods, using both anion and cation exchange chromatography. Glycerol and phosphate sugars were found in all alga samples analyzed, at concentrations between 0.11 and 22 μg g⁻¹, whereas sulfonate and sulfate sugars were only detected in three of them (0.6-7.2 μg g⁻¹). Regarding arsenic toxic species, low concentration levels of dimethylarsinic acid (DMA) (<0.9 μg g⁻¹) and generally high arsenate (As(V)) concentrations (up to 77 μg g⁻¹) were found in most of the algae studied. The results obtained are of interest to highlight the need to perform speciation analysis and to introduce appropriate legislation to limit toxic arsenic species content in these food products.     

Measurement of methyl mercury (I) and mercury (II) in fish tissues and sediments by HPLC-ICPMS and HPLC-HGAAS

A procedure for the extraction and determination of methyl mercury and mercury (II) in fish muscle tissues and sediment samples is presented. The procedure involves extraction with 5% (v/v) 2-mercaptoethanol, separation and determination of mercury species by HPLC-ICPMS using a Perkin-Elmer 3 μm C8 (33 mm × 3 mm) column and a mobile phase 3 containing 0.5% (v/v) 2-mercaptoethanol and 5% (v/v) CH₃OH (pH 5.5) at a flow rate 1.5 ml min⁻¹ and a temperature of 25 °C. Calibration curves for methyl mercury (I) and mercury (II) standards were linear in the range of 0-100 μg l⁻¹ (r² = 0.9990 and r² = 0.9995 respectively). The lowest measurable mercury was 0.4 μg l⁻¹ which corresponds to 0.01 μg g⁻¹ in fish tissues and sediments. Methyl mercury concentrations measured in biological certified reference materials, NRCC DORM - 2 Dogfish muscle (4.4 ± 0.8 μg g⁻¹), NRCC Dolt - 3 Dogfish liver (1.55 ± 0.09 μg g⁻¹), NIST RM 50 Albacore Tuna (0.89 ± 0.08 μg g⁻¹) and IRMM IMEP-20 Tuna fish (3.6 ± 0.6 μg g⁻¹) were in agreement with the certified value (4.47 ± 0.32 μg g⁻¹, 1.59 ± 0.12 μg g⁻¹, 0.87 ± 0.03 μg g⁻¹, 4.24 ± 0.27 μg g⁻¹ respectively). For the sediment reference material ERM CC 580, a methyl mercury concentration of 0.070 ± 0.002 μg g⁻¹ was measured which corresponds to an extraction efficiency of 92 ± 3% of certified values (0.076 ± 0.04 μg g⁻¹) but within the range of published values (0.040-0.084 μg g⁻¹; mean ± s.d.: 0.073 ± 0.05 μg g⁻¹, n = 40) for this material. The extraction procedure for the fish tissues was also compared against an enzymatic extraction using Protease type XIV that has been previously published and similar results were obtained. The use of HPLC-HGAAS with a Phenomenox 5 μm Luna C18 (250 mm × 4.6 mm) column and a mobile phase containing 0.06 mol l⁻¹ ammonium acetate (Merck Pty Limited, Australia) in 5% (v/v) methanol and 0.1% (w/v) l-cysteine at 25 °C was evaluated as a complementary alternative to HPLC-ICPMS for the measurement of mercury species in fish tissues. The lowest measurable mercury concentration was 2 μg l⁻¹ and this corresponds to 0.1 μg g⁻¹ in fish tissues. Analysis of enzymatic extracts analysed by HPLC-HGAAS and HPLC-ICPMS gave equivalent results.     

Determination of triazine compounds in ground water samples by micellar electrokinetic capillary chromatography

In this work, a capillary electrophoresis (CE) method for the determination of a group of eleven triazine compounds by micellar electrokinetic capillary chromatography (MEKC) with diode array detection was developed. The eleven herbicides studied were: desethylatrazin-2-hydroxy (DEA), simazine, prometon, atrazine, simetryn, ametryn, propazine, prometryn, trietazine, terbutylazine, and terbutryn The separation of these compounds was optimised as a function of buffer concentration and pH, concentration of sodium dodecyl sulphate (SDS) and voltage applied. To increase the selectivity of the separation and the resolution of the solutes, different organic solvents were tested as buffer additives, obtaining the best results when 1-propanol was used. The optimised buffer (24 mM of sodium borate, 18 mM of disodium hydrogen phosphate, 25 mM of SDS, pH 9.5, and 5% of 1-propanol) provides the best separation in terms of resolution and migration time. This method allowed the determination of these compounds at concentrations of 0.05 μg l⁻¹ in ground water samples pretreated using solid-phase extraction (SPE).     

Speciation analysis of arsenic in groundwater from Inner Mongolia with an emphasis on acid-leachable particulate arsenic

Arsenic in drinking water affects millions of people around the world. While soluble arsenic is commonly measured, the amount of particulate arsenic in drinking water has often been overlooked. We report here determination of the acid-leachable particulate arsenic and soluble arsenicals in well water from an arsenic-poisoning endemic area in Inner Mongolia, China. Water samples (583) were collected from 120 wells in Ba Men, Inner Mongolia, where well water was the primary drinking water source. Two methods were demonstrated for the determination of soluble arsenic species (primarily inorganic arsenate and arsenite) and total particulate arsenic. The first method used solid phase extraction cartridges and membrane filters to separate arsenic species on-site, followed by analysis of the individual arsenic species eluted from the cartridges and filters. The other method uses liquid chromatography separation with hydride generation atomic fluorescence detection to determine soluble arsenic species. Analysis of acidified water samples using inductively coupled plasma mass spectrometry provided the total arsenic concentration. Arsenic concentrations in water samples from the 120 wells ranged from <1 to ∼1000 μg L⁻¹. On average, particulate arsenic accounted for 39 ± 38% (median 36%) of the total arsenic. In some wells, particulate arsenic was six times higher than the soluble arsenic concentration. Particulate arsenic can be effectively removed using membrane filtration. The information on particulate and soluble arsenic in water is useful for optimizing treatment options and for understanding the geochemical behavior of arsenic in groundwater.     

Simultaneous determination of imidacloprid, carbendazim, methiocarb and hexythiazox in peaches and nectarines by liquid chromatography-mass spectrometry

A liquid chromatographic (LC) atmospheric pressure chemical ionization mass spectrometric (MS) method is described for determining imidacloprid, carbendazim, methiocarb and hexythiazox in peaches and nectarines. The samples were extracted with ethyl acetate and anhydrous sodium sulfate. Recoveries yield for spiking samples were ranged from 64±9% (R.S.D) for carbendazim to 108±14% (R.S.D.) for hexythiazox at the concentration of 0.1 mg kg⁻¹ (n=5). The correlation coefficients were greater than 0.998 over the range between 0.02 and 2 mg kg⁻¹. The limits of quantitation (LOQ) were 0.02 mg kg⁻¹ for all the pesticides. The applicability of the method to detect and quantify imidacloprid, carbendazim, methiocarb and hexythiazox was demonstrated successfully in 159 peach and nectarine samples obtained from an agricultural co-operative. The calculation of the estimated daily intakes (EDI) from these data showed that contribution of fruits to dietary intakes were much lower than acceptable daily intakes (ADI) proposed by international agencies.     

An on-line method for pressurized hot water extraction and enzymatic hydrolysis of quercetin glucosides from onions

A novel environmentally sound continuous-flow hot water extraction and enzymatic hydrolysis method for determination of quercetin in onion raw materials was successfully constructed using a stepwise optimization approach. In the first step, enzymatic hydrolysis of quercetin-3,4′-diglucoside to quercetin was optimized using a three level central composite design considering temperature (75–95 °C), pH (3–6) and volume concentration of ethanol (5–15%). The enzyme used was a thermostable β-glucosidase variant (termed TnBgl1A_N221S/P342L) covalently immobilized on either of two acrylic support-materials (Eupergit^® C 250L or monolithic cryogel). Optimal reaction conditions were irrespective of support 84 °C, 5% ethanol and pH 5.5, and at these conditions, no significant loss of enzyme activity was observed during 72 h of use. In a second step, hot water extractions from chopped yellow onions, run at the optimal temperature for hydrolysis, were optimized in a two level design with respect to pH (2.6 and 5.5), ethanol concentration (0 and 5%) and flow rate (1 and 3 mL min⁻¹) Obtained results showed that the total quercetin extraction yield was 1.7 times higher using a flow rate of 3 mL min⁻¹ (extraction time 90 min), compared to a flow rate of 1 mL min⁻¹ (extraction time 240 min). Presence of 5% ethanol was favorable for the extraction yield, while a further decrease in pH was not, not even for the extraction step alone. Finally, the complete continuous flow method (84 °C, 5% ethanol, pH 5.5, 3 mL min⁻¹) was used to extract quercetin from yellow, red and shallot onions and resulted in higher or similar yield (e.g. 8.4 ± 0.7 μmol g⁻¹ fresh weight yellow onion) compared to a conventional batch extraction method using methanol as extraction solvent.     

DNA interaction probed by evanescent wave cavity ring-down absorption spectroscopy via functionalized gold nanoparticles

Evanescent wave cavity ring-down absorption spectroscopy (EW-CRDS) is employed to study interaction and binding kinetics of DNA strands by using gold nanoparticles (Au NPs) as sensitive reporters. These Au NPs are connected to target DNA of study that hybridizes with the complementary DNA fixed on the silica surface. By the absorbance of Au NPs, the interaction between two DNA strands may be examined to yield an adsorption equilibrium constant of 2.2 × 10¹⁰ M⁻¹ using Langmuir fit. The binding efficiency that is affected by ion concentration, buffer pH and temperature is also examined. This approach is then applied to the label-free detection of the DNA mutation diseases using the sandwich hybridization assay. For monitoring a gene associated with sickle-cell anemia, the detection limit and the adsorption equilibrium constant is determined to be 1.2 pM and (3.7 ± 0.8) × 10¹⁰ M⁻¹, distinct difference from the perfectly matched DNA sequence that yields the corresponding 0.5 pM and (1.1 ± 0.2) × 10¹¹ M⁻¹. The EW-CRDS method appears to have great potential for the investigation of the kinetics of a wide range of biological reactions.     

A novel fused-silica capillary dropping mercury electrode with long drop-time and its application on determination of critical micelle concentration

A novel long drop time mercury electrode has been constructed from common fused-silica capillary (50 μm I.D., 360 μm E.D.). Proposed device provides reproducible mercury drops with typical lifetime 40-120 s. The electrode was used for a set of electrocapillary measurements aimed at determination of critical micelle concentration of anionic surfactants by a convection controlled drop-time technique. A critical micelle concentration of sodium dodecyl sulfate 5.6 ± 0.4 mmol L⁻¹ and 4.3 ± 0.4 mmol L⁻¹ were obtained in 1 mmol L⁻¹ and 5 mmol L⁻¹ phosphate buffer (pH 7.0), respectively. The values were comparable to those obtained from conductometric measurements under the same conditions (7.0 ± 0.1 mmol L⁻¹ and 5.2 ± 0.1 mmol L⁻¹, respectively) and the difference was explained in accordance with theory of hemi-micelle formation.     

Study of binding and denaturation dynamics of IgG and anti-IgG using dual color fluorescence correlation spectroscopy

In this article, we present a systematic study on IgG and Fab fragment of anti-IgG molecules using fluorescence auto- and cross-correlation spectroscopy to investigate their diffusion characteristics, binding kinetics, and the effect of small organic molecule, urea on their binding. Through our analysis, we found that the diffusion coefficient for IgG and Fab fragment of anti-IgG molecules were 37 ± 2 μm² s⁻¹ and 56 ± 2 μm² s⁻¹, respectively. From the binding kinetics study, the respective forward (k_a) and backward (k_d) reaction rates were (5.25 ± 0.25) × 10⁶ M⁻¹ s⁻¹ and 0.08 ± 0.005 s⁻¹, respectively and the corresponding dissociation binding constant (K_D) was 15 ± 2 nM. We also found that urea inhibits the binding of these molecules at 4 M concentration due to denaturation.     

Simultaneous determination of selenium and arsenic contents in different extracts of Radix Astragali by enhancement effect of ethanol in hydride generation-inductively coupled plasma-atomic emission spectrometry

A new method was developed for simultaneous determination of trace arsenic and selenium in different extracts of Radix Astragali by enhancement effect of ethanol in hydride generation-inductively coupled plasma-atomic emission spectrometry (HG-ICP-AES) with a microwave digestion system. The effects of the concentration of the hydride generating reagent (NaBH₄), ethanol concentration, different extraction methods and pre-reducing reagents on selenium and arsenic emission intensity were discussed and optimized. The contents of selenium and arsenic in different extracts (polysaccharide, amino acid, astragaloside, and water decoction,) in Radix Astragali were analyzed. The proposed method was validated by the use of two plant reference samples {poplar leaf (GBW07604) and tea (GBW07605)}. The detection limits (3σ) were 7.0 ng L^− 1 and 2.0 ng L^− 1 for Se(IV) and As(III) and relative standard deviations (RSD) were 1.8% and 2.3%, respectively. The determination of Selenium and Arsenic contents in different extracts of Radix Astragali would provide useful information for the quality control of Radix Astragali.     

Determination of total nitrogen in atmospheric wet and dry deposition samples

A microwave-assisted persulfate oxidation method followed by ion chromatographic determination of nitrate was developed for total nitrogen determination in atmospheric wet and dry deposition samples. Various operating parameters such as oxidation reagent concentrations, microwave power, and extraction time were optimized to maximize the conversion of total nitrogen to nitrate for subsequent chemical analysis. Under optimized conditions, 0.012 M K₂S₂O₈ and 0.024 M NaOH were found to be necessary for complete digestion of wet and dry deposition samples at 400 W for 7 min using microwave. The optimized extraction method was then validated by testing different forms of organic nitrogen loaded to pre-baked filter substrates and NIST SRM 1648 (urban particulate matter), and satisfactory results were obtained. In the case of wet deposition samples, standard addition experiments were performed. The suitability of the method for real-world application was assessed by analyzing a number of wet and dry deposition samples collected in Singapore during the period of March-April 2007. The organic nitrogen content was 15% (wet) and 30% (dry) of the total nitrogen. During the study period, the estimated wet fluxes for nitrate (NO₃⁻), ammonium (NH₄⁺), organic nitrogen (ON), and total nitrogen (TN) were 16.1 ± 6.5 kg ha⁻¹ year⁻¹, 11.5 ± 5.7 kg ha⁻¹ year⁻¹, 3.8 ± 1.5 kg ha⁻¹ year⁻¹and 31.5 ± 13.2 kg ha⁻¹ year⁻¹, respectively, while the dry fluxes were 2.5 ± 0.8 kg ha⁻¹ year⁻¹, 1.4 ± 0.9 kg ha⁻¹ year⁻¹, 2.3 ± 1.4 kg ha⁻¹ year⁻¹ and 7.5 ± 2.6 kg ha⁻¹ year⁻¹, respectively.     

A rapid quantitative method for humic substances determination in natural waters

A simple and rapid method was proposed for humic substances (HS) determination at microgram levels in natural waters. This assay method is based on the binding of a dye, Toluidine Blue (TB), to HS molecules to produce a dye-HS complex, which causes a decrease in absorbance at 630 nm. This method was calibrated with HS samples with up to a concentration of 40 mg L⁻¹, which covered the range of dissolved HS concentrations present in natural waters. The detection limit was 0.8 mg L⁻¹ of HS, and the relative standard deviation of 10 replicate measurements for a 20-mg L⁻¹ standard sample was 3.5%. From the Langmuir adsorption isotherm theory, the binding equilibrium constant and total number of binding sites at neutral pH were calculated to be (8.17 ± 0.42) × 10⁵ L mol⁻¹ and N of 1.45 ± 0.04 mmol g⁻¹ HS, respectively. The determination results with five water samples from lake, river and pond were consistent with those measured with the reference methods, demonstrating that this quantification method for HS determination was rapid, sensitive and feasible.     

A potential high-throughput method for the determination of lipase activity by potentiometric flow injection titrations

Potentiometric FIA titrations were performed to determine enzyme activities of lipase type B from Candida antarctica, CAL-B. Two substrates, triacetin and tributyrin were hydrolyzed in phosphate buffer solutions, and the concentration change of the base component of the buffer was titrated in a carrier solution containing hydrochloric acid and potassium chloride. The system was calibrated with butyric acid and acetic acid, respectively. FIA titration peaks were evaluated with respect to peak height and peak area. Butyric acid and acetic acid could be titrated in the buffer solution from 3 × 10⁻³ mol L⁻¹ to 0.1 mol L⁻¹. The detection limit of enzyme activity was determined to be 0.07 U mL⁻¹ (15 min reaction time) and the minimum activity was calculated to be 0.035 units corresponding to 35 nmol min⁻¹. The specific activities of lipase B for the hydrolysis of tributyrin and triacetin were determined as 16 ± 2 U mg⁻¹ and 2 ± 0.2 U mg⁻¹ (per mg commercial lipase preparation), respectively.     

Species selective preconcentration and quantification of gold nanoparticles using cloud point extraction and electrothermal atomic absorption spectrometry

The determination of metallic nanoparticles in environmental samples requires sample pretreatment that ideally combines pre-concentration and species selectivity. With cloud point extraction (CPE) using the surfactant Triton X-114 we present a simple and cost effective separation technique that meets both criteria. Effective separation of ionic gold species and Au nanoparticles (Au-NPs) is achieved by using sodium thiosulphate as a complexing agent. The extraction efficiency for Au-NP ranged from 1.01 ± 0.06 (particle size 2 nm) to 0.52 ± 0.16 (particle size 150 nm). An enrichment factor of 80 and a low limit of detection of 5 ng L⁻¹ is achieved using electrothermal atomic absorption spectrometry (ET-AAS) for quantification. TEM measurements showed that the particle size is not affected by the CPE process. Natural organic matter (NOM) is tolerated up to a concentration of 10 mg L⁻¹. The precision of the method expressed as the standard deviation of 12 replicates at an Au-NP concentration of 100 ng L⁻¹ is 9.5%. A relation between particle concentration and the extraction efficiency was not observed. Spiking experiments showed a recovery higher than 91% for environmental water samples.