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Improved chip design for integrated solid-phase microextraction in on-line proteomic sample preparation.

A recently introduced silicon microextraction chip (SMEC), used for on-line proteomic sample preparation, has proved to facilitate the process of protein identification by sample clean up and enrichment of peptides. It is demonstrated that a novel grid-SMEC design improves the operating characteristics for solid-phase microextraction, by reducing dispersion effects and thereby improving the sample preparation conditions. The structures investigated in this paper are treated both numerically and experimentally. The numerical approach is based on finite element analysis of the microfluidic flow in the microchip. The analysis is accomplished by use of the computational fluid dynamics-module FLOTRAN in the ANSYS software package. The modeling and analysis of the previously reported weir-SMEC design indicates some severe drawbacks, that can be reduced by changing the microextraction chip geometry to the grid-SMEC design. The overall analytical performance was thereby improved and also verified by experimental work. Matrix-assisted laser desorption/ionization mass spectra of model peptides extracted from both the weir-SMEC and the new grid-SMEC support the numerical analysis results. Further use of numerical modeling and analysis of the SMEC structures is also discussed and suggested in this work.

Miniaturization↗

A solid-phase microextraction fiber coated with diglycidyloxycalix[4]arene yields very high extraction selectivity and sensitivity during the analysis of chlorobenzenes in soil.

A novel fiber coated with novel sol-gel (5,11,17,23-tetra-tert-butyl-25,27-dihydroxy-26,28-diglycidyloxycalix[4]arene/hydroxy-terminated silicone oil; diglycidyloxy-C[4]/OH-TSO) was prepared for use with headspace solid-phase microextraction (HS-SPME) combined with gas chromatography (GC) and electron capture detection (ECD), which was applied in order to determine nine chlorobenzenes in soil matrices. Due to the improved fiber preparation, which increases the percentage of calixarene in the coating, the new calixarene fiber exhibits very high extraction selectivity and sensitivity to chlorine-substituted compounds. Various parameters affecting the extraction efficiency were optimized in order to maximize the sensitivity during the chlorobenzene analysis. Interferences from different soil matrices with different characteristics were investigated, and the amount extracted was strongly influenced by the matrix. Therefore, a standard addition protocol was performed on the real soil samples. The linear ranges of detection for the chlorobenzenes tested covered three orders of magnitude, and correlation coefficients >0.9976 and relative standard deviations (RSD) <8% were observed. The detection limits were found at sub-ng/g of soil levels, which were about an order of magnitude lower than those given by the commercial poly(dimethylsiloxane) (PDMS) coating for most of the compounds. The recoveries ranged from 64 to 109.6% for each analyte in the real kaleyard soil matrix when different concentration levels were determined over the linear range, which confirmed the reliability and feasibility of the HS-SPME/GC-ECD approach using the fiber coated with diglycidyloxy-C[4]/OH-TSO for the ultratrace analysis of chlorobenzenes in complex matrices.

Journal Article↗

Multielemental speciation analysis of organometallic compounds of mercury, lead and tin in natural water samples by headspace-solid phase microextraction followed by gas chromatography-mass spectrometry.

The development of a simple and rapid multielemental speciation method is described with the ultimate goal to simultaneously determine various organometallic compounds of mercury, lead and tin (inorganic mercury, methylmercury, trimethyllead, triethyllead, monobutyl-, dibutyl- and tributyltin) in natural water samples. The analytical method consists on the ethylation with NaBEt4, simultaneous headspace-solid phase microextraction (HS-SPME) of the derivatives and final gas chromatographic-mass spectrometric (GC-MS) analysis. After optimization of important process parameters, like SPME fiber coating, extraction time and extraction temperature, the analytical characteristics were evaluated. Detection limits in the low ng l(-1) level, linearity over three orders of magnitude and repeatability in the range of 3-20% were achieved for all compounds under study. The accuracy of the method in terms of average percentage recovery of the compounds in spiked river water and seawater samples was better than 90%. Finally, application of the proposed method to real natural aqueous samples enabled the simultaneous determination of all the compounds under study in seawater samples obtained from the marina area of Gijón (Asturias, Spain).

Gas Chromatography-Mass Spectrometry↗

Determination of 2,4,6-tribromoaniline in the color additives D&C Red Nos. 21 and 22 (Eosin Y) using solid-phase microextraction and gas chromatography-mass spectrometry.

The present work demonstrates the presence of an impurity, 2,4,6-tribromoaniline (TBA), in the color additives D&C Red Nos. 21 and 21 lake (21L) and describes the determination of TBA in certified lots of D&C Red Nos. 21, 21L and 22 (Eosin Y). A method was developed using solid-phase microextraction with [13C6]TBA as an internal standard followed by gas chromatography-mass spectrometry analysis. Test portions from 23 lots of US-certified color additives D&C Red Nos. 21, 21L and 22 were analyzed for TBA using the new method. These lots represent domestic (four) and foreign (four) manufacturers that requested certification for the color additives during the past 2 years. Of the test portions analyzed, 12 (52.2%) contained TBA in amounts ranging from 19.9 to 638.9 ppm with an average value of approximately 278.7 ppm. The remaining 11 (47.2%) test portions contained no detectable TBA or less than 0.01 ppm, which is the limit of quantification of the present method. The wide range of TBA levels found in lots submitted for certification suggest that the contamination with TBA may be avoided or significantly decreased through appropriate changes in the color-manufacturing process.

Aniline Compounds↗

Determination of atrazine and four organophosphorus pesticides in ground water using solid phase microextraction (SPME) followed by gas chromatography with selected-ion monitoring.

A rapid, sensitive, and convenient method is presented for the determination of atrazine and four organophosphorus pesticides (OPP) in small (10 ml) samples of ground water. Samples are initially fortified with ethion (internal standard), then extracted without organic solvent using a 65-microm thickness polydimethylsiloxane/divinylbenzene (PDMS-DVB) solid-phase microextraction (SPME) fiber. The analytes collected are thermally desorbed in a heated gas chromatographic inlet, separated using a fused-silica capillary column, and detected using a mass selective detector in its selected-ion monitoring (SIM) mode. Two independent statistical procedures were used to evaluate the detection limits, which typically range between 2 and 8 microg l(-1) for these analytes. Method performance was also evaluated using "performance evaluation" samples, in which clean authentic ground waters were fortified to known concentrations with at least two of the analytes of interest. Sample-to-sample analysis time is approximately 30 min, making the new method ideal for "quick turn" determinations.

Atrazine↗

(Z)-dodec-3-en-1-ol, a novel termite trail pheromone identified after solid phase microextraction from Macrotermes annandalei.

(Z)-dodec-3-en-1-ol was isolated and identified by GC-MS as the major component of the trail-following pheromone from whole body and sternal gland extracts of workers of the fungus-growing termite, Macrotermes annandalei (Silvestri) (Termitidae, Macrotermitinae). For the first time, this trail pheromone was also identified by using solid phase microextraction from the surface of the secretory sternal gland of workers. Bioassays showed that synthetic dodecenol induced both orientation and recruitment behavioral effects. The activity threshold of (Z)-dodec-3-en-1-ol in eliciting trail-following is similar to that of (3Z,6Z,8E)-dodeca-3,6,8-trien-1-ol in the Rhinotermitidae, but amounts of dodecenol secreted are 100 times higher than those of dodecatrienol. There is about 1 ng of (Z)-dodec-3-en-1-ol per worker. Artificial trails made of synthetic dodecenol are able to compete with natural trails in the field. The activity duration of synthetic (Z)-dodec-3-en-1-ol trails is shorter than that of trails made from whole sternal secretion of workers. Observations showed that (Z)-dodec-3-en-1-ol is probably the only major component of the trail-following pheromone of M. annandalei and that it could be associated with other compounds in a pheromonal blend providing specificity and/or stability to trails.

Journal Article↗

Preparation and application of the sol-gel-derived acrylate/silicone co-polymer coatings for headspace solid-phase microextraction of 2-chloroethyl ethyl sulfide in soil.

Three types of novel acrylate/silicone co-polymer coatings, including co-poly(methyl acrylate/hydroxy-terminated silicone oil) (MA/OH-TSO), co-poly(methyl methacrylate/OH-TSO) (MMA/OH-TSO) and co-poly(butyl methacrylate/OH-TSO) (BMA/OH-TSO), were prepared for the first time by sol-gel method and cross-linking technology and subsequently applied to headspace solid-phase microextraction (HS-SPME) of 2-chloroethyl ethyl sulfide (CEES), a surrogate of mustard, in soil. The underlying mechanisms of the coating process were discussed and confirmed by IR spectra. The selectivity of the three types of sol-gel-derived acrylate/silicone coated fibers was studied, and the BMA/OH-TSO coated fibers exhibited the highest extraction ability to CEES. The concentration of BMA and OH-TSO in sol solution was optimized, and the BMA/OH-TSO (3:1)-coated fibers possessed the highest extraction efficiency. Compared with commercially available polyacrylate (PA) fiber, the sol-gel-derived BMA/OH-TSO (3:1) fibers showed much higher extraction efficiency to CEES. Therefore, the BMA/OH-TSO (3:1)-coated fibers were chosen for the analysis of CEES in soil matrix. The reproducibility of coating preparation was satisfactory, with the RSD 2.39% within batch and 3.52% between batches, respectively. The coatings proved to be quite stable at high temperature (to 350 degrees C) and in different solvents (organic or inorganic), thus their lifetimes (to 150 times) are longer than conventional fibers. Extraction parameters, such as the volume of water added to the soil, extraction temperature and time, and the ionic strength were optimized. The linearity was from 0.1 to 10 microg/g, the limit of detection (LOD) was 2.7 ng/g, and the RSD was 2.19%. The recovery of CEES was 88.06% in agriculture soil, 92.61% in red clay, and 101.95% in sandy soil, respectively.

Acrylates↗

Analysis of strawberry volatiles using comprehensive two-dimensional gas chromatography with headspace solid-phase microextraction.

The aims of the current study were to develop an enantioselective multi-dimensional gas chromatography (GC x GC) method for the examination of strawberry volatiles and to use this method to make comparisons between the volatile profiles of different cultivars and between fresh picked and post-harvest berries of the same cultivar. Strawberry volatiles were sampled using solid-phase microextraction (SPME), and the repeatability and reproducibility of this method was examined. Semi-quantitative analysis of the volatiles was conducted using the relatively new technique of comprehensive multi-dimensional gas chromatography, using enantioselective (chiral) columns for the differentiation of analyte enantiomers. Chiral GC x GC facilitated the detection of key enantiomers in strawberry flavour. The (-)-enantiomer of 2,5-dimethyl-4-hydroxy-(2H)-furan-3-one (DMHF) and the S-enantiomer of linalool were tentatively identified as the predominant forms in both the cultivars Selva and Adina. The compounds benzaldehyde and methyl hexanoate were shown to decrease in post-harvest berries, whilst DMHF and nerolidol increased upon storage.

Chromatography, Gas↗

Simple determination of 22 organophosphorous pesticides in human blood using headspace solid-phase microextraction and gas chromatography with mass spectrometric detection.

A simple and rapid procedure for the determination of 22 organophosphorous pesticides (bromophos-ethyl, bromophos-methyl, chlorfenvinphos, chlorpyriphos, demethon-S-methylsulfon, diazinon, dichlorvos, dicrotophos, dimethoate, disulfoton, edifenphos, fenitrothion, fenthion, malathion, methidathion, mevinphos, monocrotophos, omethoate, parathion-ethyl, parathion-methyl, phosphamidon, and quinalphos) in human blood using headspace (HS) solid-phase microextraction (SPME) and gas chromatography (GC)-mass spectrometry (MS) is presented. The effects of various sample additions, incubation temperatures, absorption times, desorption times, and depths of fiber insertion into the injection port of the GC are optimized to enhance the sensitivity of the procedure. The recoveries of spiked blood samples are determined between 70% and 95% compared with samples prepared in water, and absolute recoveries are in the range between 0.1% and 19.6%. For quantitation in the single ion monitoring mode, linearity is established over concentration ranges from 0.025 to 5.0 microg/g with excellent coefficients of correlation (0.991-0.998). The detection limits are in the range between 0.01 and 0.3 microg/g. The time for analysis is 44 min per sample including extraction and GC-MS analysis. HS-SPME in combination with GC-MS is an effective method for the determination of organophosphorous pesticides in human blood and shows a great potential for use in rapid on-site analytical work, which is highly demanded in clinical and forensic toxicology.

Calibration↗

Determination of cocaine, benzoylecgonine and cocaethylene in human hair by solid-phase microextraction and gas chromatography-mass spectrometry.

The present work describes a highly precise and sensitive method developed to detect cocaine (COC), benzoylecgonine (BE, its main metabolite) and cocaethylene (CE, transesterification product of the coingestion of COC with ethanol) in human head hair samples. The method was based on an alkylchloroformate derivatization of benzoylecgonine and the extraction of the analytes by solid-phase microextraction (SPME). Gas chromatography-mass spectrometry (GC-MS) was used to identify and quantify the analytes in selected ion monitoring mode (SIM). The limits of quantification and detection (LOQ and LOD) were: 0.1 ng/mg for COC and CE, and 0.5 ng/mg for BE. Good inter- and intra-assay precision was observed. The dynamic range of the assay was 0.1-50 ng/mg. The method is not time consuming and was shown to be easy to perform.

Cocaine↗

Solid phase microextraction of aminodinitrotoluenes in tissue.

Tubifex tubifex metabolizes 2,4,6-trinitrotoluene (TNT) to 2-amino-4,6-dinitrotoluene (2ADNT) and 4-amino-2,6-dinitrotoluene (4ADNT). Elimination rates of metabolically-generated ADNTs are low compared to ADNTs absorbed directly from water, suggesting that metabolically-generated ADNTs may be bound or sequestered within tissue and therefore less available for elimination. A solid phase microextraction (SPME) technique was used to extract ADNTs from T. tubifex tissue to investigate the recalcitrance of metabolically-generated ADNTs. As SPME is a gentle, non-depletive, equilibrium sampling technique useful for measuring "available" organic compounds, we hypothesized that metabolically-generated ADNTs would be less extractable than absorbed ADNTs. T. tubifex were exposed to two scenarios to generate tissues containing absorbed ADNTs and metabolically-generated ADNTs. Tissue was then homogenized in a neutral buffer solution. Polyacrylate-coated (PA) SPME fibers were deployed and agitated in tissue homogenates to measure available ADNTs. Extractability of ADNTs from tissue containing metabolically-generated ADNTs was significantly less than expected: 50-60% based on the theoretical fiber-water partition ratio. Extractability of absorbed ADNTs was significantly higher (81-90%), and not significantly different than expected. The lower SPME extractability of metabolically-generated ADNTs may stem from the unavailability of metabolically-generated ADNTs sequestered in tissue or bound to tissue macromolecules during metabolism of TNT to ADNT. Tissue extractions using SPMEs may be able to estimate bound organic residues in tissue and serve to indicate the toxicological bioavailability of tissue-associated organic compounds.

Aniline Compounds↗

Selective determination of polychlorinated biphenyls in waste oils using liquid-liquid partition followed by headspace solid-phase microextraction and gas chromatography with atomic emission detection.

A procedure for the determination of the total content of polychlorinated biphenyls (PCBs) in industrial oil samples using gas chromatography coupled to atomic emission detection (GC-AED) is presented. Analytes were extracted from the samples using dimethylsulfoxide (DMSO), this extract was diluted with water, and PCBs were concentrated on a PDMS-DVB solid-phase microextraction (SPME) fibre using the headspace (HS) mode. Fibres were thermally desorbed for 3 min in the splitless injection port of the GC-AED system. Influence of liquid-liquid extraction conditions on the performance of the analytical procedure is presented and the need of a sample oxidation step, previous to the extraction of the PCBs with DMSO, discussed. Working under optimal conditions, quantification limits from 0.5 to 1 microg/g (total PCBs content) were obtained for several Aroclor mixtures in transformer oil samples. The repeatability of the whole sample preparation procedure (liquid-liquid partition followed by headspace SPME and GC-AED determination) ranged from 4 to 7%.

Chromatography, Gas↗

Evaluation of sample recovery of malodorous livestock gases from air sampling bags, solid-phase microextraction fibers, Tenax TA sorbent tubes, and sampling canisters.

Odorous gases associated with livestock operations are complex mixtures of hundreds if not thousands of compounds. Research is needed to know how best to sample and analyze these compounds. The main objective of this research was to compare recoveries of a standard gas mixture of 11 odorous compounds from the Carboxen/PDMS 75-microm solid-phase microextraction fibers, polyvinyl fluoride (PVF; Tedlar), fluorinated ethylene propylene copolymer (FEP; Teflon), foil, and polyethylene terephthalate (PET; Melinex) air sampling bags, sorbent 2,b-diphenylene-oxide polymer resin (Tenax TA) tubes, and standard 6-L Stabilizer sampling canisters after sample storage for 0.5, 24, and 120 (for sorbent tubes only) hrs at room temperature. The standard gas mixture consisted of 7 volatile fatty acids (VFAs) from acetic to hexanoic, and 4 semivolatile organic compounds including p-cresol, indole, 4-ethylphenol, and 2'-aminoacetophenone with concentrations ranging from 5.1 ppb for indole to 1270 ppb for acetic acid. On average, SPME had the highest mean recovery for all 11 gases of 106.2%, and 98.3% for 0.5- and 24-hr sample storage time, respectively. This was followed by the Tenax TA sorbent tubes (94.8% and 88.3%) for 24 and 120 hr, respectively; PET bags (71.7% and 47.2%), FEP bags (75.4% and 39.4%), commercial Tedlar bags (67.6% and 22.7%), in-house-made Tedlar bags (47.3% and 37.4%), foil bags (16.4% and 4.3%), and canisters (4.2% and 0.5%), for 0.5 and 24 hr, respectively. VFAs had higher recoveries than semivolatile organic compounds for all of the bags and canisters. New FEP bags and new foil bags had the lowest and the highest amounts of chemical impurities, respectively. New commercial Tedlar bags had measurable concentrations of N,N-dimethyl acetamide and phenol. Foil bags had measurable concentrations of acetic, propionic, butyric, valeric, and hexanoic acids.

Acetophenones↗

Study of the photoinduced degradation of polycyclic musk compounds by solid-phase microextraction and gas chromatography/mass spectrometry.

Polycyclic musks are widely used synthetic fragrances that have been identified during the last few years in biota samples and environmental matrices. Nevertheless, there is a lack of information concerning the photodegradation behavior of these compounds. In this work, the photoinduced degradation of six polycyclic musk compounds (Cashmeran, Celestolide, Phantolide, Galaxolide, Traseolide and Tonalide) was studied using a solid-phase microextraction (SPME) fiber as support. Musk fragrances were extracted from aqueous solutions using SPME fibers that were subsequently exposed to ultraviolet (UV) irradiation for different times. To study the degradation kinetics and to tentatively identify the photoproducts generated, gas chromatography coupled to ion trap mass spectrometry was used. Aqueous photodegradation studies were also performed. The on-fiber photodegradation approach avoids the need for further extraction processes and makes the identification of photoproducts easier, due to their higher concentration on the fibers. All musk compounds were easily photodegraded, suggesting that UV irradiation could work as a decontamination tool for these musks.

Journal Article↗

Solid-phase microextraction and gas chromatography-mass spectrometry of volatile compounds from avocado puree after microwave processing.

Microwave processing offers an alternative to blanch fruits and vegetables, since the application of high temperature and short time often results in minimum damage. An experimental design was used to investigate the effect of microwave time, pH, and avocado leaves (independent variables) on avocado flavor (response) using solid-phase microextraction (SPME)-GC-MS. Among the fully characterized flavor volatiles, 19 compounds were derived from lipid oxidation and only 4 from the avocado leaves. The main components derived from lipids were aldehydes, ketones and alcohols. Terpenoids, estragole, and 2-hexenal [E] were volatiles derived from avocado leaves. When leaves were added to fresh and microwaved avocado terpenoids and 2-hexenal [E]/hexanal ratio increased, this behavior was considered to have a positive effect on the sensorial quality of the product. From the statistical analysis of the experimental design, it was possible to determinate that the most important factors influencing the abundance of flavor compounds derived from lipids were microwave time and pH. Maximum values of these compounds were detected at high levels of microwave time and low values of pH. On the other hand, response surface of terpenoids and estragole showed an increment when microwave time and avocado leaf was increased. The region of optimum response was 30 s microwave time, pH 5.5, and 1% of avocado leaf.

Gas Chromatography-Mass Spectrometry↗

Solid-phase microextraction coupled with high performance liquid chromatography using on-line diode-array and electrochemical detection for the determination of fenitrothion and its main metabolites in environmental water samples.

The aim of this study was to develop a methodology for the analysis of the insecticide fenitrothion and its two main environmental metabolites, fenitrooxon and 3-methyl-4-nitrophenol. For this purpose, a solid-phase microextraction (SPME) method coupled to high performance liquid chromatography (LC) was optimized. Two on-line detectors, diode array (DAD) and direct current amperometrical (DCAD) were used in order to determine sensitivity and selectivity. The effects of the extraction parameters, including exposure and desorption time, pH, temperature, salt concentration and desorption mode on the extraction efficiency were studied. A satisfactory reproducibility for extractions from samples at 20 ppb-level with RSD < 12.5% (n = 10) was obtained. The calibration graphs were linear in the range of 10-1000 microg l(-1) and detection limits for the target compounds were between 1.2 and 11.8 microg l(-1) depending on which detector was used. The method was applied for determining fenitrothion and both its metabolites in river waters which run through forest areas near to aerial application of the pesticide.

Chromatography, High Pressure Liquid↗

Solid-phase microextraction in biomedical analysis.

Chromatographic methods are preferred in the analysis of organic molecules with lower molecular mass (<500 g/mol) in body fluids, i.e., the assay of drugs, metabolites, endogenous substances and poisons as well as of environmental exposure by gas chromatography (GC) and liquid chromatography (LC), for example. Sample preparation in biomedical analysis is mainly performed by liquid-liquid extraction and solid-phase extraction. However, new methods are investigated with the aim to increase the sample throughput and to improve the quality of analytical methods. Solid-phase microextraction (SPME) was introduced about a decade ago and it was mainly applied to environmental and food analysis. All steps of sample preparation, i.e., extraction, concentration, derivatization and transfer to the chromatograph, are integrated in one step and in one device. This is accomplished by the intelligent combination of an immobilized extraction solvent (a polymer) with a special geometry (a fiber within a syringe). It was a challenge to test this novel principle in biomedical analysis. Thus, an introduction is provided to the theory of SPME in the present paper. A critical review of the first applications to biomedical analyses is presented in the main paragraph. The optimization of SPME as well as advantages and disadvantages are discussed. It is concluded that, because of some unique characteristics, SPME can be introduced with benefit into several areas of biomedical analysis. In particular, the application of headspace SPME-GC-MS in forensic toxicology and environmental medicine appears to be promising. However, it seems that SPME will not become a universal method. Thus, on-line SPE-LC coupling with column-switching technique may be a good alternative if an analytical problem cannot be sufficiently dealt with by SPME.

Body Fluids↗

Headspace solid-phase microextraction gas chromatography/mass spectrometry in the analysis of the aroma constituents of 'Cannonau of Jerzu' wine.

"Cannonau of Jerzu" wine is a typical Sardinian product, obtained from only one particular grape variety. Its aroma profile is highly characteristic from the organoleptic point of view but, until now, no analytical investigation of the aroma components has been reported. This aspect has been studied by headspace solid-phase microextraction coupled with gas chromatography/mass spectrometry (GC/MS). Using this approach, 76 abundant aroma components were identified. The results thus obtained show on one hand that the terpene profile is, as expected, dependent on the quality of the grapes (in terms of selection before vintage), and, on the other, that some components are strongly dependent on the production and ageing method.

Gas Chromatography-Mass Spectrometry↗