Chemical Derivatization in Flow Analysis.

Chemical derivatization for improving selectivity and/or sensitivity is a common practice in analytical chemistry. It is particularly attractive in flow analysis in view of its highly reproducible reagent addition(s) and controlled timing. Then, measurements without attaining the steady state, kinetic discrimination, exploitation of unstable reagents and/or products, as well as strategies compliant with Green Analytical Chemistry, have been efficiently exploited. Flow-based chemical derivatization has been accomplished by different approaches, most involving flow and manifold programming. Solid-phase reagents, novel strategies for sample insertion and reagent addition, as well as to increase sample residence time have been also exploited. However, the required alterations in flow rates and/or manifold geometry may lead to spurious signals (e.g., Schlieren effect) resulting in distorted peaks and a noisy/drifty baseline. These anomalies can be circumvented by a proper flow system design. In this review, these aspects are critically discussed mostly in relation to spectrophotometric and luminometric detection.

Exploitation of a short monolithic column for in-line separation and preconcentration: Environmental friendly determination of the emerging pollutant salicylic acid in natural waters.

Salicylic acid is an emerging environmental contaminant, usually found at ng/L concentrations in natural waters. Its quantification usually involves liquid chromatography-tandem mass spectrometry, which requires complex and costly instrumentation as well as time-consuming sample pretreatment, typically involving large solvent volumes. In this work, sequential injection chromatography was exploited to develop a fast, green, cost-effective, and highly sensitive procedure for fluorimetric determination of salicylic acid in natural waters. Analyte preconcentration directly on the chromatographic column (on-column preconcentration) was exploited to improve detectability, yielding an enrichment factor of 122 (1.75 mL of sample) and takes only 8.5 min per determination. A detection limit of 20 ng/L, a linear response range from 0.06 to 5.00 µg/L, coefficients of variation lower than 3.0% (n = 10), and recoveries within 86 and 114% were estimated. The procedure was applied for the analyses of freshwater samples and results agreed with those obtained by liquid chromatography-tandem mass spectrometry at the 95% confidence level. The proposed procedure encompasses in-line concentration, isolation/separation, and detection, without the need for sample clean-up, thus minimizing the consumption of organic solvents and risk of analyte losse.

An overview of the Brazilian contributions to Green Analytical Chemistry.

Green Analytical Chemistry (GAC) is a research field that seeks for more sustainable analytical approaches to minimize the toxicity and amounts of wastes without hindering the analytical performance. This is a trend in Analytical Chemistry worldwide and because of the diversity of innovations on this subject, Brazil stands out as the third in the list of the main contributors to GAC, with ca. 11.2% of the published articles. Significant innovations and interesting applications in several fields have been presented and Brazil is continuously moving from Chemistry to Green Chemistry also in the Analytical Chemistry field. Selected contributions for sample preparation, spectro- and electroanalysis, separation techniques, chemometrics, and also procedures for point-of-care measurements are critically reviewed.

Solid-phase extractions in flow analysis.

Coupling solid-phase extraction (SPE) to flow systems has promoted a synergistic development. Whereas SPE mechanization leads to improved precision and higher sample throughput, as well as diminishes systematic errors and contamination risks, analyte concentration and separation from the sample matrix provides a remarkable impact on detectability and selectivity in flow analysis. Historical aspects, main cornerstones, tips for system design, and recent applications are critically reviewed, in the context of analyte(s) separation/concentration, sample clean-up, and release of sorbed chemical species involving both packed (e.g. mini-columns, cartridges, and disks) or fluidized (e.g. beads and magnetic materials) particles. Novel (bio)sorbents, selective synthetic materials, and stationary phases for low-pressure chromatography are also discussed. Moreover, the feasibility of SPE for sample treatment before chromatographic separation, as well as the exploitation of direct measurements on the solid phase (optosensing) are emphasized.

Flow analysis in Brazil: contributions over the last four decades.

The main contributions of Brazilian researchers to the field of flow analysis are reviewed, with an emphasis on historical developments, conceptual aspects, system design, and analytical applications. Contributions after the advent of flow injection analysis are highlighted. Novel approaches (e.g. zone merging, zone sampling, zone trapping, multi-site detection, and multi-commutation), flow modalities (e.g. monosegmented flow analysis, flow-batch analysis, multi-pumping flow analysis), as well as the pioneering implementation of different detection techniques (e.g. potentiometry, turbidimetry, flame atomic absorption spectrometry, inductively coupled plasma-optical emission spectrometry, and gravimetry) and analytical steps (e.g. titrations, membrane-less gas diffusion, and electrolytic dissolution) are highlighted. Strategies to improve analytical figures of merit and the use of the flow analyser as a tool for teaching purposes are also discussed. Contributions from Brazilian workers in the context of system miniaturization, "green" chemistry, analysis of complex samples, novel strategies and materials for in-line analyte separation/concentration, and proposals for expert systems are also highlighted. The large-scale analysis of samples of agronomical, environmental, industrial, and clinical relevance is emphasized.

A novel approach for lactose determination in cow's milk exploiting smartphone-based digital-image photometry.

Lactose, the main carbohydrate in cow's milk, may cause health problems for consumers with intolerance. Lactose determination in milk is hindered by the matrix complexity and lack of chromophore groups. Chromatography, volumetric, and spectrophotometric approaches involving chemical derivatization are time-consuming and require laborious sample preparation, which is incompatible with the high analytical demand. In this context, a novel approach is presented for lactose determination in milk exploiting smartphone-based digital-image photometry. It was based on a modification of the Benedict's method, involving formation of the violet Cu(I)/2,2'-biquinoline-4,4'-dicarboxylate (BCA) complex instead of the copper(I) oxide precipitate, aiming at improvement of sensitivity and precision. Sample pretreatment and analyte derivatization were performed in Eppendorf tubes with minimal reagent amounts and a smartphone camera was used for image acquisition under controlled conditions. Measurements were based on the RGB color system, taking channel G as the analytical response because of the complementarity with the color of the complex. Under the optimized conditions, the proposed procedure yielded a linear response up to 20 mg L-1 (r = 0.999), with a limit of detection of 1.5 mg L-1, which is compatible with determination of lactose in milk and dairy products categorized with low content of the sugar. The procedure takes less than 10 min, with a coefficient of variation of 3.0% (n = 12) and consumes as low as 160 µg Cu and 430 µg BCA per determination, thus being a more practical, fast, cost-effective, and environmental friendly analytical method.

Multi-energy calibration for determining critical metals in nickel-metal hydride battery residues by microwave-induced plasma atomic emission spectrometry.

Nickel-metal hydride batteries (NiMH) are a secondary source of high aggregate value elements, such as nickel, manganese, cobalt, and rare earths, for which recycling typically involves acid lixiviation. Designing the recycling process requires accurate determination of the elements in the leachates, which is hindered by the high complexity of the matrix. In the present study, microwave-induced plasma atomic emission spectrometry (MIP-OES) was selected as the quantitative method for elemental analysis because of its environment friendliness and cost-effectiveness. Multi-energy calibration (MEC) was also pioneeringly evaluated to circumvent matrix effects and simplify the determination of Ce, La, Ni, Co, and Mn in sulfuric acid leachates of NiMH batteries by MIP-OES. The method's analytical performance and accuracy were critically compared with external standard calibration and the standard additions method. MEC yielded superior results, with analyte recoveries within 90-110%, precision (coefficients of variation) from 1.8% to 5.8%, and limits of detection of 10, 20, 1, 400, and 60 µg kg-1 for Ni, La, Mn, Ce, and Co, respectively. The results demonstrated the ability of MEC-MIP-OES to minimize matrix effects, as well as simplify and speed up the analysis of NiMH battery leachates, which is compatible with this high-demand analytical application.

The role of 5-hydroxymethylfurfural in food and recent advances in analytical methods.

The thermal processing, storage, and transportation of foodstuffs (e.g., fruit juices, coffee, honey, and vinegar) generate 5-hydroxymethylfurfural (HMF). The food industry uses this compound as a quality marker, thus increasing the demand for fast and reliable analytical methods for its determination. This review focuses on the formation of HMF in food, its desirable and toxic effects, and recent advances in analytical methods for its determination in foodstuffs. The advantages and limitations of these analytical approaches are discussed relative to the main analytical features.

Sequential spectrofluorimetric determination of free and total glycerol in biodiesel in a multicommuted flow system.

A new procedure for spectrofluorimetric determination of free and total glycerol in biodiesel samples is presented. It is based on the oxidation of glycerol by periodate, forming formaldehyde, which reacts with acetylacetone, producing the luminescent 3,5-diacetyl-1,4-dihydrolutidine. A flow system with solenoid micro-pumps is proposed for solution handling. Free glycerol was extracted off-line from biodiesel samples with water, and total glycerol was converted to free glycerol by saponification with sodium ethylate under sonication. For free glycerol, a linear response was observed from 5 to 70 mg L(-1) with a detection limit of 0.5 mg L(-1), which corresponds to 2 mg kg(-1) in biodiesel. The coefficient of variation was 0.9% (20 mg L(-1), n = 10). For total glycerol, samples were diluted on-line, and the linear response range was 25 to 300 mg L(-1). The detection limit was 1.4 mg L(-1) (2.8 mg kg(-1) in biodiesel) with a coefficient of variation of 1.4% (200 mg L(-1), n = 10). The sampling rate was ca. 35 samples h(-1) and the procedure was applied to determination of free and total glycerol in biodiesel samples from soybean, cottonseed, and castor beans.

Large-scale flow analysis: From repetitive assays to expert analyzers.

Flow analysis is usually associated with repetitive assays, as all samples of a batch are generally handled in the same way. By exploiting computer-controlled devices (e.g. pumps, valves, injectors, commuters, and samplers), this scenario has been expanded, as a proper manifold dimensioning can be set for each sample. Initially, this dimensioning relied on previous information about each sample, added to the operating software prior to analysis of a given sample lot. Further, real-time decisions relying on feedback mechanisms started to be exploited for improving the analytical figures of merit, simplifying the laboratory management, and allowing real-time system optimization and fault detection. This is the essence of the expert flow analyzers, which involve manifold re-dimensioning by means of flow/manifold programming, often relying on multicommutation. The development of flow analysis from repetitive to real-time defined assays, the involved terminology, and trends on further development are highlighted in this review. Applications involve segmented and unsegmented flow analysis of agronomical, clinical, environmental, industrial, pharmaceutical, and geological samples.

Multi-energy calibration to circumvent matrix effects in the determination of biodiesel quality parameters by UV-Vis spectrophotometry.

Biodiesel is a sustainable fuel, which consists of a mixture of fatty acid alkyl esters obtained by the transesterification of vegetable oils or animal fats. Due to the diversity of raw materials used in its production, the biofuel can show significant differences in its physicochemical properties, which causes severe matrix effects in biodiesel analysis. In this work, a calibration strategy recently proposed for atomic spectrometry, named multi-energy calibration (MEC), was evaluated as a tool to circumvent matrix effects in molecular spectrometry. The strategy yielded more accurate results in the spectrophotometric determination of the iodine value and free glycerol in biodiesel produced from different raw materials (root mean square error of prediction, RMSEP, values of 6.0 and 8.0, respectively), than both external standard calibration (EC, RMSEP as high as 45.6) and the standard additions method (SAM, RMSEP as high as 25.3). Moreover, results agreed with those obtained by the reference procedures at the 95% confidence level, which was not observed for EC (free glycerol determination) and SAM (iodine value determination). Coefficients of variation (n = 8) and limits of detection (99.7% confidence level) were, respectively, estimated at 1.7% and 5 g I2/100 g iodine value and 3.0% and 10 mg kg-1 free glycerol in biodiesel, which are comparable to or better than those achieved using previously proposed procedures. Thus, MEC is a viable alternative for biodiesel quality control, with outstanding minimization of matrix effects, simplicity, practicality, and environmental friendliness.

Flow analysis during the 60 years of Talanta.

A parallel within the development of flow analysis and the consolidation of Talanta as one of the main journals in analytical chemistry is drawn. Influence of scientific divulgation, meeting organizations, thematic issues devoted to scientific events and Talanta awards in the recent development of flow analysis is emphasized. For didactic purposes, the discussion is focused on three 20-year periods. A scientometric overview demonstrated the consolidation of Talanta as the main journal for divulgation of recent innovations in flow analysis.

The multiple facets of flow analysis. A tutorial.

The amazing development of flow analysis has led to a loss of conceptual uniformity and to the proposals of a number of modalities, each assigned to an acronym, and this aspect may hinder further developments in the field. As any sample handling step of a flow-based analytical procedure can be accomplished in different ways, there are multiple facets associated to it. This tutorial is focused on the critical evaluation of these facets and the proposal of a novel way to present the flow analyzers, disregarding or even avoiding the need for specifying flow modalities and acronyms.

A spot test for total esters determination in sugarcane spirits exploiting smartphone-based digital images.

Cachaça is an alcoholic beverage produced from sugarcane, whose flavor and taste are related to the esters content, usually expressed as equivalent to ethyl acetate. The official method for the determination of specific esters in cachaça is based on gas chromatography with a flame ionization detector, whereas a volumetric procedure is recommended for determining the total content. Because of the high analytical demand, faster and more practical analytical procedures are required for quality control of the product. The aim of this work was to develop a spot test exploiting smartphone-based digital images for in situ determination of total esters in cachaça. The procedure was based on the reaction of the analytes with hydroxylamine, generating the corresponding hydroxamate ions, which form violet complexes with Fe(iii) in an acidic medium. Digital images were acquired under controlled illumination and converted to RGB values using the PhotoMetrix® 1.8 application. The values of channel B were taken as the analytical response because of the complementarity with the color of the reaction product. A linear response was obtained within 100-500 mg L-1 ethyl acetate, with the coefficient of variation (n = 10) and limit of detection (99.7% confidence level) estimated at 1.1% and 30 mg L-1 ethyl acetate, respectively. The procedure consumes only 1.4 mg NH2OH·HCl and 115 µg Fe(iii) and generates only 900 µL of waste per determination. The results of the proposed procedure agreed with those obtained by the reference volumetric method at the 95% confidence level.

Flow-Batch Sample Preparation for Fractionation of the Stress Signaling Phytohormone Salicylic Acid in Fresh Leaves.

Salicylic acid (SA) is an important stress signaling phytohormone and plays an essential role in physiological processes in plants. SA fractionation has been carried out batchwise, which is not compatible with the high analytical demand in agronomical studies and increases susceptibility to analytical errors. In this context, a novel flow-batch sample preparation system for SA fractionation on fresh plant leaves was developed. It was based on microwave-assisted extraction with water and conversion of the conjugated species to free SA by alkaline hydrolysis. Free and total SA were quantified by fluorimetry after separation by sequential injection chromatography in a C18 monolithic column. The proposed procedure is directly applicable to plant leaves containing up 16 mg kg-1 SA, with a limit of detection of 0.1 mg kg-1 of SA, coefficient of variation of 3.0% (n = 10), and sampling rate of 4 samples h-1. The flow-batch sample preparation system was successfully applied to SA fractionation in sugarcane, corn, and soybean leaves without clogging or increasing in backpressure. The proposed approach is simple, less time-consuming, and more environmentally friendly in comparison to batchwise procedures.

Systematic evaluation of sample preparation for fractionation of phytohormone salicylic acid in fresh leaves.

The determination of salicylic acid (SA), an important phytohormone responsible for stress signaling in plants, is of great importance in agricultural studies. However, a critical evaluation of the procedures for the extraction of the analytes and hydrolysis of the conjugated forms of SA is lacking in the literature and the available alternatives are complex, time-consuming, and laborious. In this study, the sample preparation methods for SA fractionation were critically evaluated to develop a simpler and faster alternative procedure. Microwave-assisted extractions were carried out with 2.0 g of fresh leaves and 8.0 mL of a 75% v/v ethanol:water solution at 40 °C for 10 min, followed by alkaline hydrolysis using 100 µL of 0.1 mol L-1 NaOH at 80 °C for 60 min. Free and total SA were determined in crude and hydrolyzed extracts, respectively, by fluorimetry after chromatographic separation of the sample matrix under isocratic elution (25% v/v acetonitrile/phosphate buffer) using a C18 column. Recovery experiments using methyl salicylate and acetylsalicylic acid model compounds demonstrated that the soft microwave-assisted extraction did not decompose the SA derivatives and that alkaline hydrolysis was quantitative. The proposed procedure was successfully applied for fractionation of SA in sugarcane, corn, and soybean leaves with extraction and hydrolysis yields up to 70 and 20% higher than those achieved in previously proposed approaches, respectively. The developed procedure is a simple, fast, and reliable alternative for SA fractionation in crude extracts without sample clean-up, and utilizes dilute reagents and green solvents.

Two-dimensional separation by sequential injection chromatography.

Sequential injection chromatography (SIC) is an alternative for fast chromatographic separations with low consumption of organic solvents. However, its separation capacity is restricted by the use of short chromatographic columns and the limitations for gradient elution. The present work aimed to expand the analytical capacity of SIC by exploiting a multidimensional approach with two chromatographic columns, different in their separation mechanisms, which increased the selectivity and peak resolution. The viability of the proposal was demonstrated by separation of aromatic biogenic amines (histamine, tyramine, phenylethylamine, and tryptamine), whose unidimensional separation was not achieved either by using cyanopropyl or C18 chromatographic columns. In the two-dimensional approach, the fraction of the eluate unresolved in the first dimension (containing tyramine and phenylethylamine) was collected in a sampling loop and, subsequently, inserted in the second chromatographic dimension (heart-cutting mode). Under the optimized conditions, the first chromatographic dimension was based on a cyanopropyl monolithic column and an aqueous mobile phase composed of phosphoric acid solution, pH 2.5, while the second dimension employed a C18 superficially porous particle column and a mobile phase composed of acetonitrile and phosphoric acid aqueous solution, pH 2.5 (7:93, v/v). The total analysis time was 8 min, and a resolution of 1.72 was achieved between the nearest peaks (tyramine and phenylethylamine). Linear responses were obtained within 10 and 50 mg L-1 (r > 0.997), with detection limits estimated at 2.7, 7.7, 1.9, and 0.3 mg L-1, for histamine, tyramine, phenylethylamine, and tryptamine, respectively, and a coefficient of variation of 3.0% (n = 12).

A simple and low-cost approach for microdistillation: Application to methanol determination in biodiesel exploiting smartphone-based digital images.

Although usual in analytical chemistry, separations of volatile analytes by distillation are time-consuming, whereas gas diffusion through membranes is characterized by low efficiency. These limitations directly hinder sample throughput and sensitivity. In this work, a simple and inexpensive approach is proposed for separation of volatile analytes by microdistillation. It consists of an eppendorf type tube containing the acceptor solution suspended inside a 50 mL Falcon® capped tube containing the sample. The bottom of the sample tube is submitted to controlled heating in a water bath to promote analyte volatilization to the headspace, thus favoring its collection into the acceptor solution. The efficiency of the approach was evaluated by determination of methanol in biodiesel, whose threshold limit is 0.2% (m/m) according the current legislation. Measurements were based on discoloration of an acidic potassium permanganate solution (750 µL), caused by oxidation of methanol with consequent reduction to Mn(II). The analytical measurements were based on intensity of the reflected radiation acquired as digital images by the camera of a smartphone under controlled lighting. Values of the G channel were used as analytical parameter because of the complementarity with the color of the potassium permanganate solution. Under heating at 70 °C for 40 min, the transference of methanol to the acceptor solution was estimated at (98 ±â€¯3)%. A linear response was achieved within 0.025% and 0.200% (m/m) methanol, with coefficient of variation (n = 10) and limit of detection (99.7% confidence level) estimated at 3.7% and 0.009% (m/m) respectively. The procedure requires only 100 µg of potassium permanganate and 200 µL of sample and generates as low as 950 µL of residue per determination. Because of the selectivity provided by microdistillation, the results obtained by the proposed approach agreed with the reference procedure at the 95% confidence level. The proposed approach is a simple, efficient, and cost-effective alternative for separation and determination of volatile species.

A novel spot test based on digital images for determination of methanol in biodiesel.

Methanol is used in transesterification reaction for biodiesel production, being an important quality parameter to evaluate the purity of the final product. Methanol concentrations higher than 0.20% (m/m) reduce the biofuel efficiency. The Brazilian, United States, and European standards recommend the reference method based on gas chromatography with flame ionization detection, which involves a time-consuming procedure and requires high-cost equipment. In this work, it is proposed a simple and low cost spot test based on digital images acquired with a smartphone camera for the determination of methanol in biodiesel. The procedure was based on the oxidation of methanol to formaldehyde and subsequent reaction with Schiff reagent, resulting in a violet product. Analytical signals were based on the reflected radiation measured on the G channel (RGB system), which refers to the complementary color of the reaction product. Linear responses were obtained within 20-500 mg kg-1 and 500-2000 mg kg-1 methanol for 600 and 150 µL of sample aqueous extracts, respectively. The coefficient of variation (n = 10) and the limit of detection (99.7% confidence level) were estimated as 4.8% and 10 mg kg-1 (600 µL of sample), respectively. The procedure requires only 75 µg of potassium permanganate, 6 mg of oxalic acid, and 1.5 mg of p-rosaniline and generates ca. 1.2 mL of residue per determination. The results agreed with those obtained by the reference procedure at the 95% confidence level, demonstrating that the proposed method is an alternative for routine analysis of the biofuel.

A green flow-based procedure for fluorimetric determination of acid-dissociable cyanide in natural waters exploiting multicommutation.

A flow system designed with solenoid valves is proposed for determination of weak acid dissociable cyanide, based on the reaction with o-phthalaldehyde (OPA) and glycine yielding a highly fluorescent isoindole derivative. The proposed procedure minimizes the main drawbacks related to the reference batch procedure, based on reaction with barbituric acid and pyridine followed by spectrophotometric detection, i.e., use of toxic reagents, high reagent consumption and waste generation, low sampling rate, and poor sensitivity. Retention of the sample zone was exploited to increase the conversion rate of the analyte with minimized sample dispersion. Linear response (r = 0.999) was observed for cyanide concentrations in the range 1-200 microg L(-1), with a detection limit (99.7% confidence level) of 0.5 microg L(-1) (19 nmol L(-1)). The sampling rate and coefficient of variation (n = 10) were estimated as 22 measurements per hour and 1.4%, respectively. The results of determination of weak acid dissociable cyanide in natural water samples were in agreement with those achieved by the batch reference procedure at the 95% confidence level. Additionally to the improvement in the analytical features in comparison with those of the flow system with continuous reagent addition (sensitivity and sampling rate 90 and 83% higher, respectively), the consumption of OPA was 230-fold lower.