Érica A. Souza-Silva

Sample preparation with solid phase microextraction and exhaustive extraction approaches: Comparison for challenging cases.

In chemical analysis, sample preparation is frequently considered the bottleneck of the entire analytical method. The success of the final method strongly depends on understanding the entire process of analysis of a particular type of analyte in a sample, namely: the physicochemical properties of the analytes (solubility, volatility, polarity etc.), the environmental conditions, and the matrix components of the sample. Various sample preparation strategies have been developed based on exhaustive or non-exhaustive extraction of analytes from matrices. Undoubtedly, amongst all sample preparation approaches, liquid extraction, including liquid-liquid (LLE) and solid phase extraction (SPE), are the most well-known, widely used, and commonly accepted methods by many international organizations and accredited laboratories. Both methods are well documented and there are many well defined procedures, which make them, at first sight, the methods of choice. However, many challenging tasks, such as complex matrix applications, on-site and in vivo applications, and determination of matrix-bound and free concentrations of analytes, are not easily attainable with these classical approaches for sample preparation. In the last two decades, the introduction of solid phase microextraction (SPME) has brought significant progress in the sample preparation area by facilitating on-site and in vivo applications, time weighted average (TWA) and instantaneous concentration determinations. Recently introduced matrix compatible coatings for SPME facilitate direct extraction from complex matrices and fill the gap in direct sampling from challenging matrices. Following introduction of SPME, numerous other microextraction approaches evolved to address limitations of the above mentioned techniques. There is not a single method that can be considered as a universal solution for sample preparation. This review aims to show the main advantages and limitations of the above mentioned sample preparation approaches and the applicability and capability of each technique for challenging cases such as complex matrices, on-site applications and automation.

Headspace versus Direct Immersion Solid Phase Microextraction in Complex Matrixes: Investigation of Analyte Behavior in Multicomponent Mixtures

This work aims to investigate the behavior of analytes in complex mixtures and matrixes with the use of solid-phase microextraction (SPME). Various factors that influence analyte uptake such as coating chemistry, extraction mode, the physicochemical properties of analytes, and matrix complexity were considered. At first, an aqueous system containing analytes bearing different hydrophobicities, molecular weights, and chemical functionalities was investigated by using commercially available liquid and solid porous coatings. The differences in the mass transfer mechanisms resulted in a more pronounced occurrence of coating saturation in headspace mode. Contrariwise, direct immersion extraction minimizes the occurrence of artifacts related to coating saturation and provides enhanced extraction of polar compounds. In addition, matrix-compatible PDMS-modified solid coatings, characterized by a new morphology that avoids coating fouling, were compared to their nonmodified analogues. The obtained results indicate that PDMS-modified coatings reduce artifacts associated with coating saturation, even in headspace mode. This factor, coupled to their matrix compatibility, make the use of direct SPME very practical as a quantification approach and the best choice for metabolomics studies where wide coverage is intended. To further understand the influence on analyte uptake on a system where additional interactions occur due to matrix components, ex vivo and in vivo sampling conditions were simulated using a starch matrix model, with the aim of mimicking plant-derived materials. Our results corroborate the fact that matrix handling can affect analyte/matrix equilibria, with consequent release of high concentrations of previously bound hydrophobic compounds, potentially leading to coating saturation. Direct immersion SPME limited the occurrence of the artifacts, which confirms the suitability of SPME for in vivo applications. These findings shed light into the implementation of in vivo SPME strategies in quantitative metabolomics studies of complex plant-based systems.

Direct Immersion Solid-Phase Microextraction with Matrix-Compatible Fiber Coating for Multiresidue Pesticide Analysis of Grapes by Gas Chromatography-Time-of-Flight Mass Spectrometry (DI-SPME-GC-ToFMS).

A fast and sensitive direct immersion-solid-phase microextraction-gas chromatography-time-of-flight mass spectrometry (DI-SPME-GC-ToFMS) method for the determination of multiresidue pesticides in grapes employing a PDMS-modified PDMS/DVB coating was developed utilizing multivariate approaches for optimization of the most important factors affecting SPME performance. A comprehensive investigation of appropriate internal standards using a bottom-up approach led to the selection of suitable compounds that adequately covered a range of 40 pesticides pertaining to various classes. The validated method yielded good accuracy, precision, and sensitivity and has been successfully applied to the analysis of commercial samples. With regard to the limitations of the proposed method, the DI-SPME method did not provide a satisfactory performance toward more polar pesticides (e.g., acephate, omethoate, dimethoate) and highly hydrophobic pesticides, such as pyrethroids. Despite the challenges and limitations encountered by this method, the practical aspects of the PDMS-modified coating demonstrated here create new opportunities for SPME applied in food analysis.

Capturing Plant Metabolome with Direct-Immersion in Vivo Solid Phase Microextraction of Plant Tissues

For the first time, an in vivo sampling mode of direct immersion-solid phase microextraction (DI-SPME) was employed to capture the metabolome of living plant specimens, using apple (Malus × domestica Borkh.) as a model system. Metabolites were extracted from apple tissues and introduced by thermal desorption into a comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry instrument. The feasibility of this sampling approach, based on exploitation of microextraction principles, including negligible depletion of free analyte concentrations, solventless sampling and sample preparation, and on-site compatibility, was determined in global metabolite analysis. Rather than adopting an approach of traditional sample preparation, requiring metabolism quenching and laborious sample preparation, the objective of the study was to capture the metabolome in vivo, evaluate the feasibility of the approach to provide unbiased extraction coverage, and compare analytical precision when different SPME sampling modes are employed. The potential of in vivo DI-SPME in quantitative plant metabolomics was assessed by evaluating changes in metabolic fingerprints in response to fruit maturation. The in vivo SPME sampling approach has been demonstrated as capable of sampling living systems with high reproducibility, considering that nearly 50% of hundreds of evaluated compounds included in the determination of analytical performance met the 15% RSD FDA criterion. Esters were extracted with high repeatability (% RSD for hexyl butanoate and butyl butanoate of 16.5 and 5.9, respectively, from 9 determinations in 3 apples) and found to be upregulated in response to apple fruit maturation.

Insights into the Effect of the PDMS-Layer on the Kinetics and Thermodynamics of Analyte Sorption onto the Matrix-Compatible Solid Phase Microextraction Coating

The currently presented research investigated the performance of matrix compatible PDMS-overcoated fibers (PDMS-DVB/PDMS) as compared to unmodified PDMS/DVB coatings using aqueous samples and employing a wide range of analyte polarities, molecular weights, and functionalities. In the first part of the work, a kinetic approach was taken to investigate the effect of the PDMS outer layer on the uptake rate of analytes during the mass transfer process. In short, the results can be simplified into two models: (1) the rate-limiting step is the diffusion through the coating and (2) the rate-limiting step is the diffusion through the aqueous diffusional boundary layer. For polar compounds, according to the theoretical discussion, the rate-limiting step is the diffusion through the coating; therefore, the outer PDMS layer influences the uptake rate into the matrix compatible coatings. On the other hand, for nonpolar compounds, the rate-limiting step of the uptake process is diffusion through the aqueous diffusional boundary layer; as such, the overcoated PDMS does not affect uptake rate into the matrix-compatible coatings as compared to DVB/PDMS fibers. From a thermodynamic point of view, the calculated fiber constants further corroborate the hypothesis that the additional PDMS layer does not impair the extraction phase capacity.

Coupling solid phase microextraction to complementary separation platforms for metabotyping of E. coli metabolome in response to natural antibacterial agents

IntroductionEssential oils are known to possess antimicrobial activity; thus, their use has played an important role over the years in medicine and for food preservation purposes.ObjectiveThe effect of clove oil and its major constituents as bactericidal agents on the global metabolic profiling of E. coli bacteria was assessed by means of metabolic alterations, using solid phase microextraction (SPME) as a sample preparation method coupled to complementary analytical platforms.MethodE. Coli cultures treated with clove oil and its major individual components were sampled by HS-SPME-GCxGC-ToF/MS and SPME-UPLC–MS. Full factorial design was applied in order to estimate the most effective antibacterial agent towards E. coli. Central composite design and factorial design were applied to investigate parameters influencing metabolite coverage and efficiency by SPME.ResultsThe metabolic profile, including 500 metabolites identified by LC–MS and 789 components detected by GCxGC-ToF/MS, 125 of which were identified as dysregulated metabolites, revealed changes in the metabolome provoked by the antibacterial activity of clove oil, and in particular its major constituent eugenol. Analyses of individual components selected using orthogonal projections to latent structures discriminant analysis showed a neat differentiation between control samples in comparison to treated samples in various sets of metabolic pathways.ConclusionsThe combination of a sample preparation method capable of providing cleaner extracts coupled to different analytical platforms was successful in uncovering changes in metabolic pathways associated with lipids biodegradation, changes in the TCA cycle, amino acids, and enzyme inhibitors in response to antibacterial treatment.

Exploiting the tunable selectivity features of polymeric ionic liquid-based SPME sorbents in food analysis

In this work, the performances of polymeric ionic liquid (PIL) based solid-phase microextraction (SPME) coatings were assessed for applications concerning food safety and quality. Two different polymeric ionic liquid coatings, namely poly(1-4-vinylbenzyl-3-hexadecylimidazolium) bis[(trifluoromethyl)sulfonyl] imide (poly([VBHDIM][NTf2]), PIL 1, and N,N-didecyl-N-methyl-d-glucaminium poly(2-methyl-acrylic acid 2-[1-(3-{2-[2-(3-trifluoromethanesulfonylamino-propoxy)-ethoxy]-ethoxy}-propylamino)-vinylamino]-ethyl ester) (poly([DDMGlu][MTFSI]), PIL 2, were evaluated. The PIL-based coatings were compared to commercially available SPME coatings in terms of their performance toward extraction of pesticides and fruit metabolites. The partition coefficients (Kfs) of the tested coatings were calculated, with PIL 1 demonstrating similar or better performance compared to the commercial coatings. Design of experiment (DoE) was applied to optimize the parameters that most influenced SPME extraction, and a quantitative method for determination of 5 organophosphorus pesticides was developed by using PIL-based coatings and commercial SPME fibers. Despite the thin layer of the sorbent coating, PIL 1 achieved limits of quantitation at the low part-per-billion level. Moreover, in a comparative investigation of analyte coverage carried out via HS-SPME-GCxGC-ToF/MS with grape homogenate as model matrix, excellent performances were observed for the PIL-based coatings toward the determination of fruit metabolites, demonstrating their capability towards broad extractive coverage of analytes characterized by various physicochemical properties.