Polypyrrole-coated capillary coupled to HPLC for in-tube solid-phase microextraction and analysis of aromatic compounds in aqueous samples

In-tube solid-phase microextraction (SPME) based on a polypyrrole (PPY)-coated capillary was investigated for the extraction of aromatic compounds from aqueous solutions. The PPY-coated capillary was coupled on-line to HPLC that was programmed with an autosampler to achieve automated in-tube SPME and HPLC analysis. Three groups of aromatics, including both polar and nonpolar compounds, were examined. The results demonstrated that the PPY coating had a higher extraction efficiency than the currently used commercial capillary coatings, especially for polycyclic aromatic compounds and polar aromatics due to the increasing pi-pi interactions, interactions by polar functional groups, and hydrophobic interactions between the polymer and the analytes. In addition to the functional groups in the PPY coating, which contributed to the higher extraction efficiency and selectivity toward analytes, the coating's porous surface structure,which was revealed by electron microscopy experiments, provided a high surface area that allowed for high extraction efficiency. It was found that the extraction efficiency and selectivity could be tuned by changing the coating thickness. The preliminary study of the extraction mechanism indicated that analytes were extracted onto the PPY coating mainly by an adsorption mechanism. The method was used for the extraction and analysis of both polar and nonpolar aromatics in aqueous samples.     

Automated in-tube solid-phase microextraction coupled with liquid chromatography/electrospray ionization mass spectrometry for the determination of beta-blockers and metabolites in urine and serum samples.

The technique of automated in-tube solid-phase microextraction (SPME) coupled with liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) was evaluated for the determination of beta-blockers in urine and serum samples. In-tube SPME is an extraction technique for organic compounds in aqueous samples, in which analytes are extracted from the sample directly into an open tubular capillary by repeated draw/eject cycles of sample solution. LC/MS analyses of beta-blockers were initially performed by liquid injection onto a LC column. Nine beta-blockers tested in this study gave very simple ESI mass spectra, and strong signals corresponding to [M + H]+ were observed for all beta-blockers. The beta-blockers were separated with a Hypersil BDS C18 column using acetonitrile/methanol/water/acetic acid (15:15:70:1) as a mobile phase. To optimize the extraction of beta-blockers, several in-tube SPME parameters were examined. The optimum extraction conditions were 15 draw/eject cycles of 30 microL of sample in 100 mM Tris-HCl (pH 8.5) at a flow rate of 100 microL/min using an Omegawax 250 capillary (Supelco, Bellefonte, PA). The beta-blockers extracted by the capillary were easily desorbed by mobile-phase flow, and carryover of beta-blockers was not observed. Using in-tube SPME/LC/ESI-MS with selected ion monitoring, the calibration curves of beta-blockers were linear in the range from 2 to 100 ng/mL with correlation coefficients above 0.9982 (n = 18) and detection limits (S/N = 3) of 0.1-1.2 ng/mL. This method was successfully applied to the analysis of biological samples without interference peaks. The recoveries of beta-blockers spiked into human urine and serum samples were above 84 and 71%, respectively. A serum sample from a patient administrated propranolol was analyzed using this method and both propranolol and its metabolites were detected.     

Automated in-tube solid-phase microextraction coupled with HPLC for the determination of N-nitrosamines in cell cultures

An automated in-tube solid-phase microextraction (SPME) HPLC analysis method for 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and several metabolites has been developed. NNK is one of the tobacco-specific N-nitrosamines (TSNA), which has been linked to cancers associated with the use of or exposure to tobacco products. In-tube SPME is an on-line extraction technique in which analytes are extracted and concentrated from the sample directly into a coated capillary by repeated draw/eject steps. In this study, a tailor-made polypyrrole (PPY)-coated capillary and several commercially available capillaries (capillary GC columns) were used to evaluate their extraction efficiencies for NNK and several metabolites in cell cultures. Compared with commercial capillaries that were currently used for in-tube SPME, the PPY-coated capillary showed better extraction efficiency for all of the compounds studied. After optimization of the extraction conditions, NNK and five metabolite compounds were analyzed in spiked cell cultures, confirming the applicability of the developed method. Excellent linearity was observed for all compounds (av R2 = 0.9942) and detection limits that ranged from 20 to 250 ng/mL. The average within-day and between day variations (% RSD) were 2.9 and 3.6%, respectively. This automated extraction and analysis method simplified the determination of the TSNA, requiring a total sample analysis time of only approximately 30 min.     

In-tube moleculary imprinted polymer solid-phase microextraction for the selective determination of propranolol

A molecularly imprinted polymer (MIP) material was synthesized for use as an in-tube solid-phase microextraction (SPME) adsorbent. The inherent selectivity and chemical and physical robustness of the MIP material was demonstrated as an effective stationary-phase material for in-tube SPME. An automated and on-line MIP SPME extraction method was developed for propranolol determination in biological fluids. This simplified the sample preparation process and the chromatographic separation of several beta-blocker compounds. The method developed for propranolol showed improved selectivity in comparison to alternative in-tube stationary-phase materials, overcoming the limitations of existing SPME coating materials. Preconcentration of the sample by the MIP adsorbent increased the sensitivity, yielding a limit of detection of 0.32 microg/mL by UV detection. Excellent method reproducibility (RSD < 5.0%) and column reusability (> 500 injections) were observed over a fairly wide linear dynamic range (0.5-100 microg/mL) in serum samples. To our knowledge, this is the first report on the automated application of a MIP material for in-tube SPME. The method was inexpensive, simple to set up, and simplified the choice of SPME adsorbent for in-tube extraction. The approach can potentially be extended to other MIPs for the determination of a wide range of chemically significant analytes.     

Sol-gel capillary microextraction

Sol-gel capillary microextraction (sol-gel CME) is introduced as a viable solventless extraction technique for the preconcentration of trace analytes. To our knowledge, this is the first report on the use of sol-gel-coated capillaries in analytical microextraction. Sol-gel-coated capillaries were employed for the extraction and preconcentration of a wide variety of polar and nonpolar analytes. Two different types of sol-gel coatings were used for extraction: sol-gel poly(dimethylsiloxane) (PDMS) and sol-gel poly(ethylene glycol) (PEG). An in-house-assembled gravity-fed sample dispensing unit was used to perform the extraction. The analysis of the extracted analytes was performed by gas chromatography (GC). The extracted analytes were transferred to the GC column via thermal desorption. For this, the capillary with the extracted analytes was connected to the inlet end of the GC column using a two-way press-fit fused-silica connector housed inside the GC injection port. Desorption of the analytes from the extraction capillary was performed by rapid temperature programming (at 100 degrees C/min) of the GC injection port. The desorbed analytes were transported down the system by the helium flow and further focused at the inlet end of the GC column maintained at 30 degrees C. Sol-gel PDMS capillaries were used for the extraction of nonpolar and moderately polar compounds (polycyclic aromatic hydrocarbons, aldehydes, ketones), while sol-gel PEG capillaries were used for the extraction of polar compounds (alcohols, phenols, amines). The technique is characterized by excellent reproducibility. For both polar and nonpolar analytes, the run-to-run and capillary-to-capillary RSD values for GC peak areas remained under 6% and 4%, respectively. The technique also demonstrated excellent extraction sensitivity. Parts per quadrillion level detection limits were achieved by coupling sol-gel CME with GC-FID. The use of thicker sol-gel coatings and longer capillary segments of larger diameter (or capillaries with sol-gel monolithic beds) should lead to further enhancement of the extraction sensitivity.     

Facile Preparation of SiO(2)/TiO(2) Composite Monolithic Capillary Column and Its Application in Enrichment of Phosphopeptides

A novel SiO(2)/TiO(2) composite monolithic capillary column was prepared by sol-gel technology and successfully applied to enrich phosphopeptides as a metal oxide affinity chromatography (MOAC) material. For the monolith preparation, tetramethoxysilane (TMOS) and tetrabutoxytitanium (TBOT) were used as silica and titania source, respectively, and glycerol was introduced to attenuate the activity of titanium precursor, which provided a mild synthetic condition. The prepared monolith was characterized by energy dispersive X-ray spectroscopy (EDX) and X-ray diffraction (XRD). The results revealed an approximate 1/2 molar ratio of titanium to silica as well as an atom-scale homogeneity in the framework. The scanning electron microscopy (SEM) results demonstrated an excellent anchorage between the column and the inner capillary wall, and nitrogen adsorption-desorption experiments showed a bimodal porosity with a narrow mesopore distribution around 3.6 nm. The prepared monolith was then applied for selective enrichment of phosphopeptides from the digestion mixture of phosphoproteins and bovine serum albumin (BSA) as well as human blood serum, nonfat milk, and egg white using an in-tube solid phase microextraction (SPME) system. Our results showed that SiO(2)/TiO(2) composite monolithic capillary column could efficiently enrich the phosphopeptides from complex matrixes. To the best of our knowledge, this is the first attempt for preparing the silica-metal composite monolithic capillary column, which offers the promising application of the monolith on phosphoproteomics study.     

固相萃取-双柱气相色谱法测定水中多种有机磷农药残留

针对单一气相色谱柱定性分析易出现假阳性误判的情况,采用固相萃取-气相色谱双柱技术测定环境水体中多种有机磷农药的残留量。水样中的有机磷农药通过HLB固相萃取小柱富集,经洗脱浓缩后,用DB-35MS和HP-5毛细管气相色谱柱、火焰光度检测器测定。方法的平均加标回收率为86.1%～118%,相对标准偏差为3.2%～6.8%,最低检出限为20～40ng/L,可满足环境水体中痕量有机磷农药残留的分析需要。     

In-tube solid-phase microextraction coupled to capillary LC for carbamate analysis in water samples

Recently, the on-line sample preparation technique, intube solid-phase microextraction (SPME), was successfully implemented with a Hewlett-Packard 1100 HPLC system for analysis of carbamates in water samples. This paper describes the coupling of in-tube SPME to capillary LC and explores its utility as a sample preparation method in that format, relative to conventional LC. The Hewlett-Packard HPLC system was upgraded to a capillary LC system using commercially available accessories from LC Packings. The combination of in-tube SPME with a capillary LC system was expected to build on the merits of both in-tube SPME and the capillary LC to generate a sensitive method with an easy, effective, and efficient sample preparation. Due to the relatively large effective injection volume of the in-tube SPME technique (30-45 microL), on-column focusing was employed in order to achieve good chromatographic efficiency. Excellent sensitivity was achieved with very good method precision. For all carbamates studied, the RSD of retention time was between 0.5 and 0.8% under 4 microL/min microgradient conditions. The RSD of peak area counts was between 1.5 and 4.6%. The detection limits for all carbamates studied were less than 0.3 microg/L and, for carbaryl, just 0.02 microg/L (20 ppt). Compared with the conventional in-tube SPME/LC method, the LODs were lowered for carbaryl, propham, methiocarb, promecarb, chlorpropham, and barban, by factors of 24, 45, 42, 81, 62, and 56, respectively. The optimized method was successfully applied to the analysis of carbamates in surface water samples.     

Thin-film microextraction

The properties of a thin sheet of poly(dimethylsiloxane) (PDMS) membrane as an extraction phase were examined and compared to solid-phase microextraction (SPME) PDMS-coated fiber for application to semivolatile analytes in direct and headspace modes. This new PDMS extraction approach showed much higher extraction rates because of the larger surface area to extraction-phase volume ratio of the thin film. Unlike the coated rod formats of SPME using thick coatings, the high extraction rate of the membrane SPME technique allows larger amounts of analytes to be extracted within a short period of time. Therefore, higher extraction efficiency and sensitivity can be achieved without sacrificing analysis time. In direct membrane SPME extraction, a linear relationship was found between the initial rate of extraction and the surface area of the extraction phase. However, for headspace extraction, the rates were somewhat lower because of the resistance to analyte transport at the sample matrix/headspace barrier. It was found that the effect of this barrier could be reduced by increasing either agitation, temperature, or surface area of the sample matrix/headspace interface. A method for the determination of PAHs in spiked lake water samples was developed based on the membrane PDMS extraction coupled with GC/MS. A linearity of 0.9960 and detection limits in the low-ppt level were found. The reproducibility was found to vary from 2.8% to 10.7%.     

Dynamic liquid-liquid-liquid microextraction with automated movement of the acceptor phase

A new dynamic liquid-liquid-liquid microextraction procedure, with the automated movement of acceptor phase (LLLME/AMAP) to facilitate mass transfer, was developed in this study. Four compounds, 3-nitrophenol, 4-nitrophenol, 3,4-dinitrophenol, and 2,4-dichlorophenol, were used as model compounds to be preconcentrated from water samples. The extraction involved filling a 2-cm length of hollow fiber with 4 muL of acceptor solution using a conventional microsyringe, followed by impregnation of the pores of the fiber wall with 1-octanol. The fiber was then immersed in 4 mL of aqueous sample solution. The analytes in the sample solution were extracted into the organic solvent and then back-extracted into the acceptor solution. During extraction, the acceptor phase was repeatedly moved in and out of the hollow fiber channel and the syringe controlled by a syringe pump. Separation and quantitative analyses were then performed by using high-performance liquid chromatography. The results indicated that up to 400-fold enrichment of the analytes could be obtained under the optimized conditions. The enrichment factors were two times those of static liquid-liquid-liquid microextraction. Good repeatabilities (RSD values below 9.30%) were obtained. The calibration linear range was from 10 to 1000 ng/mL with the square of the correlation coefficient (r2) >0.9916. Detection limits were in the range of 0.45-0.98 ng/mL. In addition, as compared with the previously reported dynamic three-phase microextraction in which there was no relative movement between the acceptor and the organic phase (which is not conducive to effective mass transfer), this new method shows much higher extraction efficiency. All these results suggest that this new dynamic LLLME/AMAP technique could be a better alternative to the previous LLLME for the extraction of analytes from aqueous samples.     

Separation and determination of polycyclic aromatic hydrocarbons by solid phase microextraction/cyclodextrin-modified capillary electrophoresis

A sensitive method for the determination of polycyclic aromatic hydrocarbons (PAHs) by solid phase microextraction coupled with cyclodextrin (CD)-modified capillary electrophoresis (CE) using UV detection has been developed. A glass fiber was prepared and used for absorbing 16 EPA priority PAHs from diluted samples until equilibrium was reached. After the glass fiber was connected to a separation capillary via an adapter, the absorbed analytes were directly released into the CE buffer stream, and electrophoretic separation was effected using a 50 mM borate, pH 9.2, buffer containing 35 mM sulfobutyloxy-β-CD, 10 mM methyl-β-CD, and 4 mM α-CD. Separation was effected since neutral PAHs differentially partitioned between the neutral and charged CD phases. Under 30 kV applied potential, separation was achieved in less than 15 min with high resolution and number of theoretical plates. Pyrene as low as 8 ppb was detected, while the highest limit of detection was 75 ppb for acenaphthene. Very satisfactory reproducibility with respect to migration time and peak area was obtained for repetitions using the same separation capillary and adapter, where only the extraction fiber was discarded after each analysis.     

Penicillium digitatum immobilized on pumice stone as a new solid phase extractor for preconcentration and/or separation of trace metals in environmental samples

This study presents a column solid phase extraction procedure based on column biosorption of Cu(II), Zn(II) and Pb(II) ions on Penicillium digitatum immobilized on pumice stone. The analytes were determined by flame atomic absorption spectrometry (FAAS). The optimum conditions such as: pH values, amount of solid phase, elution solution and flow rate of sample solution were evaluated for the quantitative recovery of the analytes. The effect of interfering ions on the recovery of the analytes has also been investigated. The recoveries of copper, zinc and lead under the optimum conditions were found to be 97+/-2, 98+/-2 and 98+/-2%, respectively, at 95% confidence level. For the analytes, 50-fold preconcentration was obtained. The analytical detection limits for Cu(II), Zn(II) and Pb(II) were 1.8, 1.3 and 5.8 ng mL(-1), respectively. The proposed procedure was applied for the determination of copper, zinc and lead in dam water, waste water, spring water, parsley and carrot. The accuracy of the procedure was checked by determining copper, zinc and lead in standard reference tea samples (GBW-07605).     

Liquid-phase microextraction of phenolic compounds combined with on-line preconcentration by field-amplified sample injection at low pH in micellar electrokinetic chromatography.

This paper describes a novel method that applies field-amplified sample injection (FASI) in micellar electrokinetic chromatography (MEKC) with a low pH background electrolyte (BGE). Six phenolic compounds prepared in water or NaOH solution were used as the test analytes. Sample was injected electrokinetically after the introduction of a plug of water. During the injection, the water plug was pumped out of the capillary inlet by the electroosmotic flow, and the phenolic anions migrated very quickly in the direction of the outlet. When the anions reached the boundary between the water plug and BGE, they were neutralized and ceased moving. Thereafter, MEKC was initiated for the separation. This on-line preconcentration method could be conveniently coupled with a liquid-liquid-liquid microextraction procedure, in which a hollow fiber was used as an extraction solvent support to extract the analytes from the water sample. The acceptor phase consisted of 8 mM NaOH. After extraction, the extract was analyzed directly by MEKC, as described.     

Absorption of hydrophobic compounds into the poly(dimethylsiloxane) coating of solid-phase microextraction fibers: high partition coefficients and fluorescence microscopy images

The use of solid-phase microextraction with poly(dimethylsiloxane) (PDMS)-coated glass fibers for the extraction and analysis of hydrophobic organic analytes is increasing. The literature on this topic is characterized by large discrepancies in partition coefficients and an uncertainty of whether highly hydrophobic analytes are retained by absorption into the fiber coating or by adsorption to the fiber surface. We applied a new method, which minimizes the impact of experimental artifacts, to determine PDMS water partition coefficients of 17 hydrophobic analytes including chlorinated benzenes, PCBs, PAHs, and p,p'-DDE. These partition coefficients are several orders of magnitude higher than some reported values. Two observations strongly suggest that the retention of hydrophobic organic substances is governed by partitioning into the PDMS coating. (1) The partition coefficients are proportional with octanol/water partition coefficients. (2) The fluorescence of fluoranthene was observed to be homogeneously distributed within the polymer coating when studied by means of fluorescence microscopy. Implications of these findings for the application of solid-phase microextraction with respect to potential detection limits, with respect to biomimetic extraction, and with respect to measurements in multicompartment systems are discussed.     

Continuous electrophoretic purification of individual analytes from multicomponent mixtures

Individual analytes can be isolated from multicomponent mixtures and collected in the outlet vial by carrying out electrophoretic purification through a capillary column. Desired analytes are allowed to migrate continuously through the column under the electric field while undesired analytes are confined to the inlet vial by application of a hydrodynamic counter pressure. Using pressure ramping and buffer replenishment techniques, 18% of the total amount present in a bulk sample can be purified when the resolution to the adjacent peak is approximately 3. With a higher resolution, the yield could be further improved. Additionally, by periodically introducing fresh buffer into the sample, changes in pH and conductivity can be mediated, allowing higher purity (>or=99.5%) to be preserved in the collected fractions. With an additional reversed cycle of flow counterbalanced capillary electrophoresis, any individual component in a sample mixture can be purified providing it can be separated in an electrophoresis system.     

SFE-GC with quantitative transfer of the extraction effluent to a megabore capillary column

Supercritical fluid extraction was directly coupled with gas chromatography such that the entire extraction effluent was directed onto the GC column. A sealed connection between the restrictor and the column ensured that no loss of analyte could occur. The elevated pressure generated in the column itself resulted in a reduced flow rate through the column, making efficient trapping possible. Metal-jacketed capillary columns were used as they can tolerate the elevated pressures required. In addition to maximizing sensitivity, very practical extraction parameters were possible. For a 25-min extraction, with a 2.2 mL/min flow rate, and the column at 40 degrees C, analytes as volatile as C-14 were trapped and separated. Trapping efficiency improved further with shorter extraction times, mild cooling of the column, or lower flow rates.     

Kinetic calibration for automated headspace liquid-phase microextraction

The kinetics of the absorption and desorption of analytes for headspace liquid-phase microextraction (HS-LPME) were studied. It was found that the desorption of analytes from the extraction phase into the sample matrix is isotropic to the absorption of the analytes from the sample matrix into the extraction phase under the same conditions. This therefore allows for the calibration of absorption using desorption. Calibration was accomplished by exposing the extraction phase, which contained a standard, to the sample matrix. The information from the desorption of the standard, such as time constant a, could be directly used to estimate the concentration of the target analyte in the sample matrix. This new kinetic calibration method for headspace LPME was successfully used to correct the matrix effects in the BTEX analysis of an orange juice sample. In this study, the headspace LPME techniques were successfully fully automated, for both static and dynamic methods, with the CTC CombiPal autosampler. All operations of headspace LPME, including sample transfer and agitation, filling of extraction solvent, exposing the solvent in the headspace, withdrawing the solvent to syringe and introducing the extraction phase into injector, were autoperformed by the CTC autosampler. The fully automated headspace LPME technique is more convenient and improved the precision and sensitivity of the method. This automated dynamic headspace LPME technique can be also used to obtain the distribution coefficient between the sample matrix (aqueous or another solution) and the extraction phase (1-octanol or another solvent). The distribution coefficient between 1-octanol and orange juice, at 25 degrees C, was obtained with this technique.     

Hybrid field-assisted solid-liquid-solid dispersive extraction for the determination of organochlorine pesticides in tobacco with gas chromatography

A novel one-step sample preparation technique termed hybrid field-assisted solid-liquid-solid dispersive extraction (HF-SLSDE) was developed in this study. A simple glass system equipped with a condenser was designed as an extraction vessel. The HF-SLSDE technique was a three-phase dispersive extraction approach. Target analytes were extracted from the sample into the extraction solvent enhanced by the hybrid field. Meanwhile, the interfering components were adsorbed by dispersing sorbent. No cleanup step preceded chromatographic analysis. The efficiency of the HF-SLSDE approach was demonstrated in the determination of organochlorine pesticide (OCP) residues in tobacco with a gas chromatography-electron capture detector (GC-ECD). Various operation conditions were studied systematically. Low detection limits (0.3-1.6 μg/kg) and low quantification limits (1.0-4.5 μg/kg) were achieved under the optimized conditions. The recoveries of OCPs ranged from 70.2% to 118.2%, with relative standard deviations of <9.6%, except for the lowest fortification level. Because of the effect of the hybrid field, HF-SLSDE showed significant predominance compared with other extraction techniques. The dispersing sorbent with good cleanup ability used in this study was also found to be a microwave absorption medium, which could heat the nonpolar extraction solvent under microwave irradiation. Different microstructures of tobacco samples before and after extractions demonstrated the mechanism of HF-SLSDE was based on an explosion at the cell level. According to the results, HF-SLSDE was proved to be a simple and effective sample preparation method for the analysis of pesticide residues in solid samples and could potentially be extended to other nonpolar target analytes in a complex matrix.     

Affinity monolith-integrated poly(methyl methacrylate) microchips for on-line protein extraction and capillary electrophoresis

Immunoaffinity monolith pretreatment columns have been coupled with capillary electrophoresis separation in poly(methyl methacrylate) (PMMA) microchips. Microdevices were designed with eight reservoirs to enable the electrically controlled transport of selected analytes and solutions to carry out integrated immunoaffinity extraction and electrophoretic separation. The PMMA microdevices were fabricated reproducibly and with high fidelity by solvent imprinting and thermal bonding methods. Monoliths with epoxy groups for antibody immobilization were prepared by direct in situ photopolymerization of glycidyl methacrylate and ethylene glycol dimethacrylate in a porogenic solvent consisting of 70% 1-dodecanol and 30% cyclohexanol. Antifluorescein isothiocyanate was utilized as a model affinity group in the monoliths, and the immobilization process was optimized. A mean elution efficiency of 92% was achieved for the monolith-based extraction of fluorescein isothiocyanate (FITC)-tagged human serum albumin. FITC-tagged proteins were purified from a contaminant protein and then separated electrophoretically using these devices. The developed immunoaffinity column/capillary electrophoresis microdevices show great promise for combining sample pretreatment and separation in biomolecular analysis.     

Optimization of the coating procedure for a high-throughput 96-blade solid phase microextraction system coupled with LC-MS/MS for analysis of complex samples

Biocompatible C18-polyacrylonitrile (PAN) coating was used as the extraction phase for an automated 96-blade solid phase microextraction (SPME) system with thin-film geometry. Three different methods of coating preparation (dipping, brush painting, and spraying) were evaluated; the spraying method was optimum in terms of its stability and reusability. The high-throughput sample preparation was achieved by using a robotic autosampler that enabled simultaneous preparation of 96 samples in 96-well-plate format. The increased volume of the extraction phase of the C18-PAN thin film coating resulted in significant enhancement in the extraction recovery when compared with that of the C18-PAN rod fibers. Various factors, such as reusability, reproducibility, pH stability, and reliability of the coating were evaluated. The results showed that the C18-PAN 96-blade SPME coating presented good extraction recovery, long-term reusability, good reproducibility, and biocompatibility. The limits of detection and quantitation were in the ranges of 0.1-0.3 and 0.5-1 ng/mL for all four analytes.