Microwave assisted biocidal extraction is an alternative method to measure microbial biomass of carbon from cultivated and non-cultivated soils.

Developing simple and cost-effective methods for soil microbial biomass carbon (MBC) measurement eases routine laboratory analysis and enables large numbers of soil samples to be measured in a relatively short period of time. Thus, the objective of this study was to develop a microwave-assisted biocidal-extraction (MWE) method which does not employ CHCl3 as biocide and K2SO4 as C-extractor, to estimate MBC. First, the microorganisms of soil samples are killed using microwave (MW) irradiation at energy level of 800 J g-1 soil as biocide followed by microwave irradiation extraction (MWE) at 562 W (120 J g-1 soil for 1 min), using deionized water as solvent. Microbial biomass of carbon from two contrasting soils microwaved with 80, 100, and 140 J g-1 soil did not differ from those obtained by using the chloroform fumigation-extraction (CFE) method with 0.5 mol L-1 K2SO4 as extractant. To evaluate the robustness of the MWE method, twenty-six soil samples, from cultivated and non-cultivated areas, with clay contents from 70-690 g kg-1, organic carbon from 5.52 to 50.82 g C kg-1 and pH values from 3.9 to 6.8 were analyzed for MBC using MWE and CFE methods. There was a linear regression (MW = - 17.87 + 0.92*K2SO4; R2 = 0.705; p < 0.001) between MWE and CFE. The biocidal microwave-assisted extraction method using 120 J g-1 soil for 1 min is a cleaner method for evaluating MBC, because it does not require chloroform, potassium sulfate salt and takes a shorter time to extract a set of soil samples.

Development of an analytical method for identification of Aspergillus flavus based on chemical markers using HPLC-MS.

Aspergillus flavus is a filamentous fungus found in nature and characterized by the production of bright and colourful colonies. It grows on different substrates, producing secondary metabolites and, if present in foodstuffs, can be a source of health problems for humans and animals, as well as causing economic losses. Traditional methods for fungal identification are based on morphological characteristics, requiring specialists and being very time-consuming. The development of analytical alternatives might have advantages such as greater efficiency, more reproducibility and be less time-consuming. Thus, a qualitative analytical method to detect Aspergillus flavus in food samples, based on the identification of fungal chemical markers by HPLC-MS, was developed. The method comprises methanol extraction followed by HPLC-MS analysis, and was able to identify 14 fungus secondary metabolites, namely aflatoxin B1, aflatoxin G1, aspergillic acid, aspyrone, betaine, chrysogine, deacetyl parasiticolide A, flufuran, gregatin B, hydroxysydonic acid, nicotinic acid, phomaligin A, spinulosin and terrein.

Toxicity assessment from electro-coagulation treated-textile dye wastewaters by bioassays.

In this study the pollutant removal from a textile dyeing wastewater has been investigated by using the electro-coagulation technique with iron electrodes. In order to obtain optimal values of the system state variables, a 3(3) full factorial experimental design was applied. The electro-coagulation (EC) process response was evaluated on the basis of COD removal and decolourization values. The electrolysis time and density current were statistically significant for the COD removal and decolourization. Based on the lettuce seeds (Lactuca sativa) and brine shrimp (Artemia salina), the lowest toxicity level was achieved in 5 min of electrolysis time. Due to the remaining high toxicity level above 30 min of electrolysis time, the EC process is not adequate to be used in a single effluent treatment, suggesting that this electrochemical process of up to 5 min could be used as part of a complete effluent treatment system.

Simultaneous determination of the textile dyes in industrial effluents by first-order derivative spectrophotometry.

A first-order derivative spectrophotometric method for the simultaneous determination of three textile dyes, Procion Yellow HE4R, Procion Red HE7B and Remazol Black 5 (RB5), has been developed. The effects of pH, heating and ionic strength of the solution on the absorption spectra of the dyes were investigated. The wavelengths selected for the measures of the derivative signals of HE4R (395 nm), HE7B (604 nm) and RB5 (659 nm) presented these coefficients of linear correlation: 0.9978, 0.9992 and 0.9999, and these detection limits: 0.180, 0.317 and 0.0233 mg L(-1), respectively. The reliability and reproducibility of the method were tested and showed recovery values of 95.7 to 109%. The proposed method was applied for the determination of dyes in binary and ternary mixtures of textile effluents and showed an estimate of the loss of dyes for the effluents between 6.67 and 28.9%.

Development of a green chromatographic method for determination of fat-soluble vitamins in food and pharmaceutical supplement.

A green chromatographic analytical method for determination of fat-soluble vitamins (A, E, D3 and K1) in food and pharmaceutical supplement samples is proposed. The method is based on the modification of a C18 column with a 3.00% (w/v) sodium dodecyl sulphate (SDS) aqueous solution at pH 7 (0.02 mol L(-1) phosphate buffer solution) and in the usage of the same surfactant solution as mobile phase with the presence of 15.0% (v/v) butyl alcohol as an organic solvent modifier. After the separation process, the vitamins are detected at 230 nm (K1, D3 and E), 280 nm (A, E, D3 and K1) and 300 nm (K1, D3 and E). The chromatographic procedure yielded precise results (better than 5%) and is able to run one sample in 25 min, consuming 1.5 g of SDS, 90 mg of phosphate and 7.5 mL of butyl alcohol. When the flow rate of the mobile phase is 2 mL min(-1) the retention times are 4.0, 9.6, 13.0 and 22.7 min for D3, A, E and K1 vitamins, respectively; and all peak resolutions are higher than 2. The analytical curves present the following linear equations: area=6290+34852 (vitamin A), R2=0.9998; area=4092+36333 (vitamin E), R2=0.9997; area=-794+30382 (vitamin D3) R2=0.9998 and area=-7175+82621 (vitamin K1), R2=0.9996. The limits of detection and quantification for vitamins A, E, D(3) and K(1) were estimated for a test pharmaceutical vitamin supplement sample as 0.81, 1.12, 0.91 and 0.83 mg L(-1) and 2.43, 3.36, 2.73 and 2.49, respectively. When the proposed method was applied to food and pharmaceutical sample analysis, precise results were obtained (R.S.D.<5% and n=3) and in agreement with those obtained by using the classical chromatographic method that uses methanol and acetonitrile as mobile phase. Here, the traditional usage of toxic organic solvent as mobile phase is avoided, which permits to classify the present method as green.

Spectrophotometric determination of Blue Procion HEGN in effluents of textile industry exploiting the dye aggregation effect and flow injection analysis.

A new spectrophotometric method involving flow injection analysis and textile dye aggregation effect is proposed. The method is based on the aggregation effect of Blue Procion HEGN at pH 3, which relocates its maximum absorption wavelength from 620 to 776 nm, avoiding the interference of other blue textile dyes. For this task, a simple and robust flow injection system was designed, which became a very stable analytical method. When the system was applied to Blue Procion determination in effluent of textile industry, precise results were observed (RSD < 2% within 1.0 and 5.0 mg l(-1) HEGN). The analytical frequency was 80 measurements per hour; the analytical curve was linear from 1.0 to 5.0 mg l(-1) HEGN; the detection limit considering three times the standard deviation of the blank solution (n = 10) was estimated as 0.03 mg l(-1) HEGN; and recoveries between 95% and 105% were found. The system consumes 20 mg of sodium citrate and 125 microl of the sample per determination. No baseline drift was observed during extended (5 h) operation periods.

Development of a green chromatographic method for determination of colorants in food samples.

A green chromatographic analytical method for determination of Tartrazine, Brilliant Blue and Sunset Yellow in food samples is proposed. The method is based on the modification of a C18 column with a 0.25% (v/v) Triton X-100 aqueous solution at pH 7 and in the usage of the same surfactant solution as mobile phase without the presence of any organic solvent modifier. After the separation process on the chromatographic column, the colorants are detected at 430, 630 and 480 nm, respectively. The chromatographic procedure yielded precise results and is able to run one sample in only 8 min, consuming 15.0mg of Triton X-100 and 38.8 mg of phosphate. When the flow rate of the mobile phase is 1 ml min(-1) the retention times are 2.1, 3.6 and 7.0 min for Tartrazine, Brilliant Blue and Sunset Yellow, respectively; and all peak resolutions are ca. 2. The analytical curves present the following linear equations: area=7.44 10(5)+2.71 10(5) [Tartrazine] (R=0.998, n=7); area=1.09 10(5)+3.75 10(5) [Brilliant] (R=0.9995, n=7) and area=-7.34 10(4)+2.33 10(5) [Sunset] (R=0.998), n=7) and, the limits of detection for Tartrazine, Brilliant Blue and Sunset Yellow were estimated as 0.125, 0.080 and 0.143 mg l(-1). When the proposed method is applied to food samples analysis, precise results are obtained (R.S.D.<5%, n=3) and in agreement with those obtained by using the classical spectrophotometric method. The traditional usage of organic solvent as mobile phase in HPLC is not used here, which permits to classify the present method as green.

Correlation of animal diet and fatty acid content in young goat meat by gas chromatography and chemometrics.

The meat fatty acids (FA) profiles of caprines submitted to different dietary treatments were determined by gas chromatography. The data were treated by Chemometrics to consider all variables together. The contents of saturated FA (SFA), monounsaturated FA (MUFA), polyunsaturated FA (PUFA), omega-3 (n-3) FA, and omega-6 (n-6) FA in 32 samples were analyzed. PUFA:SFA and n-6:n-3 ratios were also considered. The multivariate methods of hierarchical cluster analysis (HCA) and principal component analysis (PCA) were used to analyze the experimental results. HCA can group samples according to their basic composition, and PCA can explain the relationship among the dietary treatments according to the meat fatty acid composition. Treatment 1 presented the highest n-6 FA concentration, PUFA:SFA, and n-6:n-3 ratios, and the lowest MUFA and n-3 concentrations. Opposite results were observed for treatment 4. Treatments 2 and 3 were highly similar with differences mainly in SFA and MUFA concentrations.

Exploiting the bead injection concept for sequential determination of copper and mercury ions in river-water samples.

A procedure involving bead-injection concept and sequential determination of copper and mercury ions in river-water samples is proposed. The method is based on the solid-phase extraction of both metal ions on the same beads surface (Chelex 100 resin) and in their subsequent reaction with the colorimetric reagents (APDC and Dithizone for copper and mercury ions, respectively). For this task, a resin mini-column is established in the optical path by the selection, introduction and trapping of a defined volume of the Chelex-100 resin beads suspension in the flow system. The passage of the sample solution through the resin mini-column promotes the sorption of Cu(II) ions and, making the APDC colorimetric reagent flows through the beads, the formation of the coloured complex on the solid phase surface occurs. The absorbance of the formed APDC-Cu complex is then monitored at 436nm and the spent beads are discarded. Packing another resin mini-column in the flow cell and repeating the concentration step it is possible to carried out the mercury determination by using Dithizone as reagent. The absorbance of the Dithizone-Hg complex is monitored at 500nm. After each measurement, the spent beads are wasted and a new portion of fresh one is trapped in the system, letting it ready for the next measurement. The bead injection system is versatile and can be used to concentrate different sample volumes, which permits the determination of a wide range of copper and mercury ions concentrations. When the sample-selected volumes are 100 and 1000mul the analytical ranges were 5.0 up to 500.0mugl(-1) and 2.5 up to 30.0mugl(-1) for Cu(II) and Hg(II) ions, respectively. Under these conditions, the detection limit was estimated as 0.63 and 0.25mugl(-1) for copper and mercury ions determination. The system consumes 2mg of Chelex 100 resin beads, 0.20mg of APDC or 1.25mg of Dithizone per determination and the traditional organic solvent extraction methodology, normally used in connection with APDC and Dithizone reagents, is not used here which permits to classify the present method as green.

Bead-injection determination of total mercury in river water samples.

A bead-injection system is proposed for total mercury determination in river-water samples. The procedure is based on the introduction of a defined quantity of a resin suspension in the flow system. The selected beads are packed inside of a flow cell and the formed resin mini-column constitutes the optical path. The sample volume is then selected, and its passage by the mini-column allows retention of the mercury ions on the surfaces of the beads. The introduction of a spectrophotometric reagent in the flow system leads to the formation of a colored Hg-dithizone complex on the surface of the bead, which is spectrophotometricaly monitored. The spent beads are directed to waste, allowing the system to become ready to process another sample. The proposed system handles about 20 measurements per hour, consuming 1000 microl of the sample, 1 mg of Chelex 100 resin and 1.25 microg of Dithizone per determination. When 1000 microl of the sample is injected, a linear analytical curve is obtained (A = 0.0052[Hg] + 0.1028, from 0 up to 30 microg l(-1), R2 = 0.995); the detection limit is estimated to be 0.9 microg l(-1). The results are precise, r.s.d. < 9%; spiked sample recoveries within 91.2 and 109% are found.

A new strategy for exploiting ion exchange in sequential injection analysis: in-line phytic acid separation/determination in foods as an example.

A novel strategy for exploiting ion exchange in sequential injection systems is proposed. The procedure is based on the selection of a defined volume of a resin suspension, which is introduced and packed in the analytical path, establishing a resin mini-column in the system. The passage of a selected sample volume through the resin mini-column leads to the retention of the analyte, while the sample matrix is discarded. The analyte is eluted during the passage of the eluant/reagent by the packed beads, being the analytical signal monitored (absorbance) in the liquid phase. The beads are then aspirated back to the holding coil and directed to a recovery flask, linked at the selection valve; then the system is ready to begin a new cycle. With the proposed strategy, the main characteristics of the sequential injection system are kept as any new artifact is added to the manifold and system reconfiguration is not required. The feasibility of the approach is demonstrated by the phytic acid determination in food samples. For this specific application, AG1-X8 was selected as ion exchanger, and a solution containing Cl- and Fe(III)-salicylate complex was used as eluant and spectrophotometric reagent.