Methodology for soil respirometric assays: Step by step and guidelines to measure fluxes of trace gases using microcosms.

This methodology is proposed to measure the fluxes of trace gases among microcosms and the atmosphere. As microcosm respiration we include both aerobic and anaerobic respiration, which may results in CO2, CH4, NO, N2O, N2, H2S and H2 fluxes. Its applicability includes the assessment of products biodegradability and toxicity, the effect of treatments and products on greenhouse gases fluxes, and the mineralization of organic fertilizers. A step by step procedure; the complementary parameters and good practices that might be taken into account to perform a microcosm experiment; and the tools nowadays available that could be useful in this respirometric methodology are presented. We included a spreadsheet with calculus examples. Samples were taken at 1; 30; 60 and 90 min after closing the microcosms to determine the gases fluxes. The dilution effect was negligible, as we present. Besides CO2, we have successfully quantified the fluxes of CH4 and N2O from the microcosms in a broad range of concentrations. This method is useful in technical and scientific studies, for instances to test new products and improve the understanding of microbial processes, respectively. •Simple materials are required to set up the microcosm.•Examples of (pre) treatments are given regarding water availability, fertilizer doses, pH adjustment and nutrients amendments.•The method was suitable to directly measure multiple trace gases fluxes, either produced or consumed during microcosm respiration.

4-hydrazinobenzoic acid as a derivatizing agent for aldehyde analysis by HPLC-UV and CE-DAD.

Aldehydes are relevant analytes in a wide range of samples, in particular, food and beverages but also body fluids. Hydrazines can undergo nucleophilic addition with aldehydes or ketones giving origin to hydrazones (a group of stable imines) that can be suitably used in the identification of aldehydes. Herein, 4-hydrazinobenzoic acid (HBA) was, for the first time, used as the derivatizing agent in analytical methodologies using liquid chromatography aiming the determination of low-molecular aldehydes. The derivatization reaction was simultaneously performed along with the extraction process, using gas-diffusion microextraction (GDME), which resulted in a clean extract containing the HBA-aldehyde derivates. The corresponding formed imines were determined by both high-performance liquid chromatography (LC) with UV spectrophotometric detection (HPLC-UV) and capillary electrophoresis with diode array detection (CE-DAD). HBA showed to be a rather advantageous derivatization reagent due to its stability, relatively high solubility in water and other solvents, high selectivity and sensibility, reduced impurities, simple preparation steps and applicability to different separation and/or different detection techniques. Limits of detections (LODs) of the optimized methodologies (in terms of time and pH among other experimental variables) were all below 0.5 mg L-1, using both instrumental techniques. Furthermore, for the first time, the HBA-aldehyde derivatives were analyzed by LC with mass spectrometry (LC-MS), demonstrating the possibility of identification by MS of each compound. The developed methodologies were also successfully applied in the analysis of formaldehyde and acetaldehyde in several alcoholic beverages. This was also the first time GDME was combined with CE, showing that it can be a valuable sample preparation tool for electrophoresis, in particular by eliminating the interference of ions and inorganic constituents present in the samples.

Mixture-designed electrolytes for the MEKC separation of natural and synthetic steroids.

In this work, the separation of 11 natural and synthetic steroids was studied using MEKC electrolytes modified by property-selected organic solvents: ethanol, ACN, and THF. The interplay between electrophoretic behavior and structural features was disclosed and the effects of organic modifiers to modulate retention and to alter selectivity were discussed in terms of system linear solvation energy relationships (LSER). The LSER indicated the total organic solvent percentage in the electrolyte as a major parameter to control retention and as a minor contribution, the hydrogen bond acidity. By evaluating the electropherograms obtained from mixture-designed electrolytes, a favorable separation condition for all solutes was achieved in ca. 25 min with an electrolyte composed of 20 mmol/L sodium tetraborate at pH 9.4, 20 mmol/L SDS and 20% EtOH (0.80% CV for migration time and 2.5% CV for peak area, n = five consecutive injections). The applicability of the proposed separation condition was demonstrated by the inspection of estrogens in urine sample (puberty stage).

Determination of antiretroviral agents in human serum by capillary electrophoresis.

In this work, a simple and rapid electrokinetic chromatography method for the simultaneous separation of different protease inhibitors (indinavir, ritonavir, saquinavir, nelfinavir), nucleoside reverse transcriptase inhibitors (stavudine, zidovudine, didanosine) and non-nucleoside reverse transcriptase inhibitors (nevirapine, efavirenz) was developed. The analyses were performed in a 75 microm i.d. uncoated fused-silica capillary with 48.5 cm length (effective length of 40 cm) using a running buffer consisting of 20 mmol L(-1) sodium dodecyl sulfate, 10 mmol L(-1) sodium tetraborate, 30% acetonitrile and 5% ethanol. Samples were injected hydrodynamically by applying 50 mbar pressure during 6 s. All analytes were separated within 10 min with a voltage of 20 kV. The proposed method was validated for zidovudine, didanosine and efavirenz in human serum. Serum samples were prepared using a solid-phase extraction procedure (Waters Oasis HLB cartridges). For quantitative purposes, stavudine was chosen as the internal standard (IS). Method validation parameters were determined revealing good migration time repeatability (<0.7% RSD) and peak area repeatability (<1.2% RSD). Intra- and inter-day precisions were less than 1.7% and 4.4% RSD, respectively. Matrix matching analytical curves for each drug were linear in the 1.0-20.0 microg mL(-1) interval (r > 0.998). Limits of detection (LOD) were in range of 0.3-0.5 microg mL(-1). The extraction recoveries were higher than 90% with exception of efavirenz, which was 77.4%. Based on the performance characteristics, the proposed method was found suitable for the determination of zidovudine, didanosine and efavirenz in serum samples.

Determination of volatile corrosion inhibitors by capillary electrophoresis.

In this work, a capillary electrophoresis (CE) method using indirect UV detection (214nm) for the simultaneous determination of monoethanolamine (MEA), diethanolamine (DEA), triethanolamine (TEA), diethylethanolamine (DEEA), monocyclohexylamine (MCHA) and dicyclohexylamine (DCHA) in water/ethanol extracts of wrapping materials containing volatile corrosion inhibitors (VCIs) was described. A running buffer consisting of 0.010 molL(-1) imidazole, 0.010 molL(-1) 2-hydroxyisobutyric acid (HIBA) and 0.010 molL(-1) 18-crown-6 ether enabled separation of the analytes in less than 7 min. A few method validation parameters were determined revealing good migration time repeatability (<0.7% RSD) and area repeatability (< 1.8% RSD). Limits of detection were in the range of 0.52-1.54 mg L(-1). Recovery values were in the range of 94.8-100.9%. The methodology was successfully applied to the analysis of three commercial products (VCI treated paper, foam and plastic). The concentration of amines in these materials varied from 0.050 to 22.3% (w/w).

Analysis of low molecular weight aldehydes in air samples by capillary electrophoresis after derivatization with 4-hydrazinobenzoic acid.

This work reports the analysis of selected aldehydes in air samples using capillary electrophoresis (CE). The method is based on the reaction of aldehydes with 4-hydrazinobenzoic acid (HBA) to give the corresponding hydrazones with maximum absorbance at 290 nm. Under optimized CE conditions, the HBA derivatives of four carbonyls (formaldehyde, acetaldehyde, propionaldehyde, and acrolein) were completely separated from one another, in less than 6 min, using a pH 9.3 tetraborate buffer at 0.040 mol L(-1) concentration as background electrolyte. A few method validation parameters were determined revealing good migration time repeatability (< 1.5% CV) and area repeatability (< 2% CV), excellent linearity (50-300 microg/L, r > 0.996) and adequate sensitivity for environmental applications. The limits of detection with respect to each single aldehyde were in the range of 2.7-8.8 ng L(-1). The methodology was applied to the determination of aldehydes indoors. Samples were collected in HBA impregnated octadecylsilica cartridges, at different times during the day. The most abundant carbonyls in the samples were acetaldehyde followed by formaldehyde, with estimated peak concentrations of 4.3 and 2.9 ppbv, respectively.

Indirect determination of chloride and sulfate ions in alcohol fuel by capillary electrophoresis.

A capillary zone electrophoresis method using indirect UV detection for the analysis of chloride and sulfate in alcohol fuel samples was developed. The anions were analyzed in less than 3 min using an electrolyte containing 10 mmol l(-1) chromate and 0.75 mmol l(-1) hexamethonium bromide (HMB) as electroosmotic flow modifier. Coefficients of variation were better than 0.6% for migration time ( n=10) and between 2.05 and 2.82% for peak area repeatabilities. Analytical curves of peak area versus concentration in the range of 0.065-0.65 mg kg(-1) for chloride and 0.25-4.0 mg kg(-1) for sulfate were linear with coefficients of correlation higher than 0.9996. The limits of detection for sulfate and chloride were 0.033 and 0.041 mg kg(-1), respectively. Recovery values ranged from 85 to 103%. The method was successfully applied for the quantification of sulfate and chloride in five alcohol fuel samples. The concentration of sulfate varied from 0.45 to 3.12 mg kg(-1). Chloride concentrations were below the method's LOD.

Determination of low-aliphatic aldehydes indoors by micellar electrokinetic chromatography using sample dissolution manipulation for signal enhancement.

This work describes a novel approach for the analysis of selected aldehydes (formaldehyde, acetaldehyde, propionaldehyde, and acrolein) and acetone in environmental samples using micellar electrokinetic chromatography (MEKC). The method is based on the reaction of carbonyl compounds with 3-methyl-2-benzothiazoline hydrazone (MBTH) that gives an azine intermediate with maximum absorbance at 216 nm. A systematic evaluation of sample dissolution medium was conducted as a means to enhancing sensitivity. In the best condition, samples were dissolved in 0.030 mol.L(-1) tetraborate solution. This condition presented enhancement factors in the range of 35-54 for the aldehydes under investigation, computed as the improvement of the concentration limits of detection (LODs) with reference to the sample dissolved in pure water. The running buffer was 0.020 mol.L(-1) tetraborate, pH 9.3, containing 0.050 mol.L(-1) sodium dodecyly sulfate (SDS). The overall methodology presented several advantages over established methods for aldehydes. Worthy mentioning that MBTH is available in high purity degree, dispensing laborious reagent purification procedures. A few method validation parameters were determined revealing good migration time repeatability (< 2.5% coefficient of variation, CV) and area repeatability (< 4% CV), excellent linearity (20-120 micro g/L, r > 0.995) and adequate sensitivity for environmental applications. The LODs with respect to each single aldehyde were in the range of 0.54-4.0 micro g.L(-1) and 11 micro g.L(-1) for acetone. The methodology was applied to the determination of aldehydes indoors. Samples were collected in an impinger flask containing 0.05% MBTH solution, at a flow rate of 0.80 L.min(-1), during 2.5 h, at different times during the day. The most abundant carbonyls in the samples were acetone, followed by formaldehyde and acetaldehyde, with estimate peak concentrations of 452, 5.2 and 2.2 ppbv, respectively.