Determination of fluoroquinolones in eggs using in-tube solid-phase microextraction coupled to high-performance liquid chromatography

A simple, rapid, and sensitive method using in-tube solid-phase microextraction (in-tube SPME) based on poly(methacrylic acid–ethylene glycol dimethacrylate) (MAA–EGDMA) monolith coupled to HPLC with fluorescence and UV detection was developed for the determination of five fluoroquinolones (FQs). Ofloxacin (OFL), norfloxacin (NOR), ciprofloxacin (CIP), enrofloxacin (ENRO), and sarafloxacin (SARA) can be enriched and determined in the spiked eggs and albumins. CIP/ENRO in eggs and albumins of ENRO-treated hens were also studied using the proposed method. Only homogenization, dilution, and centrifugation were required before the sample was supplied to the in-tube microextraction, and no organic solvents were consumed in the procedures. Under the optimized extraction conditions, good extraction efficiency for the five FQs was obtained with no matrix interference in the process of extraction and the subsequent chromatographic separation. The detection limits (S/N=3) were found to be 0.1–2.6 ng g⁻¹ and 0.2–2.4 ng g⁻¹ in whole egg and egg albumin, respectively. Good linearity could be achieved over the range 2–500 ng mL⁻¹ for the five FQs with regression coefficients above 0.9995 in both whole egg and albumin. The reproducibility of the method was evaluated at three concentration levels, with the resulting relative standard deviations (RSDs) less than 7%. The method was successfully applied to the analysis of ENRO and its primary metabolite CIP in the eggs and albumins of ENRO-treated hens.     

On-line analysis of carbonyl compounds with derivatization in aqueous extracts of atmospheric particulate PM10 by in-tube solid-phase microextraction coupled to capillary liquid chromatography

A new device for carbonyl compounds based on coupling on-line and miniaturizing both, sample pretreatment and chromatographic separation, is reported. Two capillary columns, a GC capillary column (95% methyl-5% phenyl substituted backbone, 70 cm × 0.32 mm i.d., 3 μm film thickness) in the injection valve for in-tube solid-phase microextraction (IT-SPME) and a Zorbax SB C18 (150 mm × 0.5 mm i.d., 5 μm particle diameter) LC capillary column were employed. Different combinations of IT-SPME and derivatization using 2,4-dinitrophenylhydrazine (DNPH) were examined for mixtures containing 15 carbonyl compounds (aliphatic, aromatic and unsaturated aldehydes and ketones). A screening analysis of aqueous extracts of atmospheric particulate PM(10) was carried out. Moreover, the possibility of coupling IT-SPME and conventional liquid chromatography is also tested. Derivatization solution and IT-SPME coupled to capillary liquid chromatography provided the best results for achieving the highest sensitivity for carbonyl compounds in atmospheric particulate analysis. Detection limits (LODs) using a photodiode array detector (DAD) were ranged from 30 to 198 ng L(-1), improving markedly those LODs reported by conventional SPME-LC-DAD.     

In-tube solid-phase microextraction coupled by in valve mode to capillary LC-DAD: Improving detectability to multiresidue organic pollutants analysis in several whole waters

A simple and fast capillary chromatographic method has been developed to identify and quantify organic pollutants at sub-ppb levels in real water samples. The major groups of pesticides (organic halogens, organic phosphorous, and organic nitrogen compounds), some hydrocarbons (polycyclic aromatic hydrocarbons), phthalates and some phenols such as phenol and bisphenol A (endocrine disruptors) were included in this study. The procedure was based on coupling, in-tube solid-phase microextraction (IT-SPME) by using a conventional GC capillary column (95% methyl–5% phenyl substituted backbone, 80 cm × 0.32 mm i.d., 3 μm film thickness) in the injection valve to capillary liquid chromatography with diode array detection. A comparative study between the IT-SPME manifold and a column-switching device using a C₁₈ column (35 mm × 0.5 mm i.d., 5 μm particle size) has been performed. The IT-SPME procedure was optimal, it allows reaching limits of detection (LODs) between 0.008 and 0.2 μg/L. No matrix effect was found and recoveries between 70 and 116% were obtained. The precision of the method was good, and the achieved intra- and inter-day variation coefficients were between 2 and 30%. This procedure has been applied to the screening analysis of 28 compounds in whole waters from several points of the Mediterranean coast (Valencia Community, Spain).     

Fully automated in-tube solid-phase microextraction for liquid samples coupled to gas chromatography

An online device is described in which analytes are extracted from a liquid sample by means of in-tube solid-phase microextraction (in-tube SPME), pulse released by rapid heating, and transferred to a gas chromatograph in a fully automated way. Switching of the sample and gas flows as well as the heating of the extraction tube and the valves is controlled by a remote computer system. Results obtained for river water and for aqueous standard solutions of phenanthrene are presented and are compared to the performance of standard SPME.     

管内固相微萃取-气相色谱法联用在线测定水中有机物

将管内固相微萃取与气相色谱法结合，建立了水中痕量有机物的在线分析装置。采用毛细管气相色谱柱作为萃取柱，水样中的分析物在萃取柱中被萃取浓缩，溶剂解吸后的样品通过阀切换和柱内进样技术直接由载气携带进入气相色谱柱。采用OV-1萃取柱，对水样中的5种芳烃的富集倍数为34～85。     

Liquid-phase deposition of silica nanoparticles into a capillary for in-tube solid-phase microextraction coupled with high-performance liquid chromatography

A silica nanoparticle (NP)-deposited capillary fabricated by liquid-phase deposition (LPD) and modified with octadecyl groups was introduced for in-tube solid-phase microextraction coupled to high-performance liquid chromatography with UV detection (in-tube SPME–HPLC). The resultant capillary (60 cm × 50 μm I.D.) was demonstrated to be of higher extraction capacity by comparing with an octadecyl-grafted bare capillary and an octadecyl-grafted silica-coated capillary that was prepared by sol–gel chemistry. Two groups of compounds, endocrine disruptors and polycyclic aromatic hydrocarbons, were used as model analytes to further evaluate extraction capacity of the silica NP-deposited capillary, and its reproducibility and stability was also investigated. The extraction time profiles were monitored for all the chemicals, and their limits of detection were calculated to be in the range of 0.42–0.78 and 0.034–0.19 ng/mL with RSD values of peak area less than 4.6%.     

Octyl-type monolithic columns of 530 microm i.d. for capillary liquid chromatography

A novel monolithic capillary column (530 microm i.d.) was prepared for capillary liquid chromatography (CLC) by in situ copolymerization of octyl methacrylate (MAOE) and ethylene dimethacrylate (EDMA) in the presence of a porogen solvent containing 1-propanol, 1,4-butanediol, and water with azobisisobutyronitrile as the initiator. The influences of the contents of the porogen solvent, EDMA and the various concentration ratios of 1-propanol to 1,4-butanediol in the polymerization mixture on the morphology, porosity, globule size, stability and column efficiency were investigated. The morphology and pore size distribution of monolithic capillary columns were characterized by SEM and mercury intrusion porosimetry, respectively. Chromatographic evaluations of the columns were performed under CLC mode. The results showed that good permeability and stability can be obtained under optimal experimental conditions. The separation results of some acid, neutral and basic analytes demonstrated the hydrophobicity and low affinity to basic analytes of the new column. Three metal ions, i.e. Mg(II), Zn(II) and Cd(II) were also separated under ion-pair mode on the new monolithic capillary column and the results were acceptable.     

Determination of fluoxetine and norfluoxetine enantiomers in human plasma by polypyrrole-coated capillary in-tube solid-phase microextraction coupled with liquid chromatography-fluorescence detection

A sensitive, selective, and reproducible in-tube polypyrrole-coated capillary (PPY) solid-phase microextraction and liquid chromatographic method for fluoxetine and norfluoxetine enantiomers analysis in plasma samples has been developed, validated, and further applied to the analysis of plasma samples from elderly patients undergoing therapy with antidepressants. Important factors in the optimization of in-tube SPME efficiency are discussed, including the sample draw/eject volume, draw/eject cycle number, draw/eject flow-rate, sample pH, and influence of plasma proteins. Separation of the analytes was achieved with a Chiralcel OD-R column and a mobile phase consisting of potassium hexafluorophosphate 7.5 mM and sodium phosphate 0.25 M solution, pH 3.0, and acetonitrile (75:25, v/v) in the isocratic mode, at a flow rate of 1.0 mL/min. Detection was carried out by fluorescence absorbance at Ex/Em 230/290 nm. The multifunctional porous surface structure of the PPY-coated film provided high precision and accuracy for enantiomers. Compared with other commercial capillaries, PPY-coated capillary showed better extraction efficiency for all the analytes. The quantification limits of the proposed method were 10 ng/mL for R- and S-fluoxetine, and 15 ng/mL for R- and S-norfluoxetine, with a coefficient of variation lower than 13%. The response of the method for enantiomers is linear over a dynamic range, from the limit of quantification to 700 ng/mL, with correlation coefficients higher than 0.9940. The in-tube SPME/LC method can therefore be successfully used to analyze plasma samples from ageing patients undergoing therapy with fluoxetine.     

Fully automated analysis of estrogens in environmental waters by in-tube solid-phase microextraction coupled with liquid chromatography-tandem mass spectrometry

A simple, rapid and sensitive method for the determination of five estrogens, estrone, 17beta-estradiol, estriol, ethynyl estradiol, and diethylstilbestrol, was developed using a fully automated method consisting of in-tube solid-phase microextraction (SPME) coupled with liquid chromatography-tandem mass spectrometry (LC/MS/MS). These estrogens were separated within 8 min by HPLC using an XDB-C8 column and 0.01% ammonia/acetonitrile (60/40, v/v) at a flow rate of 0.2 mL/min. Electrospray ionization conditions in the negative ion mode were optimized for MS/MS detection of the estrogens. The optimum in-tube SPME conditions were 20 draw/eject cycles of 40 microL of sample using a Supel-Q PLOT capillary column as an extraction device. The extracted compounds were easily desorbed from the capillary by passage of the mobile phase, and no carryover was observed. Using the in-tube SPME LC/MS/MS method, good linearity of the calibration curve (r > or = 0.9996) was obtained in the concentration range from 10 to 200 pg/mL for all compounds examined. The limits of detection (S/N= 3) of the five estrogens examined ranged from 2.7 to 11.7 pg/mL. The in-tube SPME method showed 34-90-fold higher sensitivity than the direct injection method (5 microL injection). This method was applied successfully to the analysis of environmental water samples without any other pretreatment and interference peaks. Several surface water and wastewater samples were collected from the area around Asahi River, and estriol was detected at 35.7 pg/mL in the effluent of a sewage treatment plant. The recoveries of estrogens spiked into river waters were above 86%, except for estriol, and the relative standard deviations were below 0.9-8.8%.     

A simple and rapid method for simultaneous determination of benzoic and sorbic acids in food using in-tube solid-phase microextraction coupled with high-performance liquid chromatography

A simple, rapid and sensitive on-line method for the simultaneous determination of benzoic and sorbic acids in food was developed by coupling in-tube solid-phase microextraction (SPME) to high-performance liquid chromatography (HPLC) with UV detection. The diethylamine-modified poly(glycidyl methacrylate-co-ethylene dimethacrylate) monolithic capillary selected as the extraction medium exhibited a high extraction capability towards benzoic and sorbic acids. To obtain optimum extraction performance, several in-tube SPME parameters were investigated, including pH value, inorganic salt, and the organic solvent content of the sample matrix. After simple dilution with 0.02 mol/L phosphate solution (pH 4.0), carbonated drink, juice drink, sauce and jam samples could be directly injected for extraction. For succade samples, a small amount of acetonitrile was required to extract analytes prior to dilution and subsequent extraction. The linearity of the method was investigated over a concentration range of 5–20000 ng/mL for both analytes, and the correlation coefficients (R ² values) were higher than 0.999. The detection limits for benzoic and sorbic acids were 1.2 and 0.9 ng/mL, respectively. The method reproducibility was tested by evaluating the intra- and interday precisions; relative standard deviations of less than 4.4 and 9.9%, respectively, were obtained. Recoveries of compounds from spiked food samples ranged from 84.4 to 106%. The developed method was shown to be suitable for the routine monitoring of benzoic and sorbic acids in various types of food samples.     

Selective capillary coating materials for in-tube solid-phase microextraction coupled to liquid chromatography to determine drugs and biomarkers in biological samples: A review

In-tube solid-phase microextraction (in-tube SPME) coupled with high performance liquid chromatography (HPLC) or liquid chromatography coupled to mass spectrometry (LC-MS) successfully determines drugs or biomarkers in biological samples by direct sample injection or by simple sample treatment. This technique uses a capillary column as extraction device. Several capillaries (wall-coated open tubular, sorbent-packed, porous monolithic rods, or fiber-packed) with unique phases have been developed and evaluated, aiming to improve the efficiency and selectivity of the in-tube SPME-LC technique. This review describes new developments and applications occurred in recent years, and discusses future trends with emphasis on new extraction devices and current technology used for the synthesis of selective sorbents for bioanalysis, such as (i) polypyrrole, (ii) restricted-access materials, (iii) immunosorbents, (iv) molecular imprinting polymers, (v) monolithic polymers, and (vi) bi-functional materials.     

Fast analysis of soy isoflavones by high-performance liquid chromatography with monolithic columns

A fast method using high-performance liquid chromatography based on two monolithic columns has been developed for the simultaneous determination of isoflavones extracted from soybeans and derived foods. The 12 main isoflavones were resolved in 10 min in two coupled monolithic columns working at 35 °C using a elution gradient of acidified water (0.1% acetic acid) and methanol (0.1% acetic acid) at a flow rate of 5 mL min⁻¹. Retention time and relative area standard deviations were below 1% for all isoflavones. The method developed was successfully applied to several soy food samples and spiked samples. Total isoflavone concentration in sampled soy foods ranged from 34.28 mg L⁻¹ to 4.29 mg g⁻¹.     

Determination of polycyclic aromatic hydrocarbons in food samples by automated on-line in-tube solid-phase microextraction coupled with high-performance liquid chromatography-fluorescence detection

A simple and sensitive automated method, consisting of in-tube solid-phase microextraction (SPME) coupled with high-performance liquid chromatography-fluorescence detection (HPLC-FLD), was developed for the determination of 15 polycyclic aromatic hydrocarbons (PAHs) in food samples. PAHs were separated within 15 min by HPLC using a Zorbax Eclipse PAH column with a water/acetonitrile gradient elution program as the mobile phase. The optimum in-tube SPME conditions were 20 draw/eject cycles of 40 μL of sample using a CP-Sil 19CB capillary column as an extraction device. Low- and high-molecular weight PAHs were extracted effectively onto the capillary coating from 5% and 30% methanol solutions, respectively. The extracted PAHs were readily desorbed from the capillary by passage of the mobile phase, and no carryover was observed. Using the in-tube SPME HPLC-FLD method, good linearity of the calibration curve (r > 0.9972) was obtained in the concentration range of 0.05–2.0 ng/mL, and the detection limits (S/N = 3) of PAHs were 0.32–4.63 pg/mL. The in-tube SPME method showed 18–47 fold higher sensitivity than the direct injection method. The intra-day and inter-day precision (relative standard deviations) for a 1 ng/mL PAH mixture were below 5.1% and 7.6% (n = 5), respectively. This method was applied successfully to the analysis of tea products and dried food samples without interference peaks, and the recoveries of PAHs spiked into the tea samples were >70%. Low-molecular weight PAHs such as naphthalene and pyrene were detected in many foods, and carcinogenic benzo[a]pyrene, at relatively high concentrations, was also detected in some black tea samples. This method was also utilized to assess the release of PAHs from tea leaves into the liquor.     

Determination of perfluorooctanoic acid and perfluorooctane sulfonate by automated in-tube solid-phase microextraction coupled with liquid chromatography-mass spectrometry

We have developed a simple, rapid, and sensitive method for the determination of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) by on-line in-tube solid-phase microextraction (SPME) coupled with liquid chromatography-mass spectrometry (LC-MS). PFOA and PFOS were separated within 10 min by high-performance liquid chromatography using an Inertsil ODS-3 column and 10 mM ammonium acetate/methanol (35/65, v/v) as a mobile phase at a flow rate of 0.25 mL min⁻¹. Electrospray ionization conditions in the negative ion mode were optimized for MS detection of PFOA and PFOS. The optimum in-tube SPME conditions were 20 draw/eject cycles with a sample size of 40 μL using a CP-Pora PLOT amine capillary column as the extraction device. The extracted compounds could be desorbed easily from the capillary by passage of the mobile phase, and no carryover was observed. Using the in-tube SPME LC-MS method, good linearity of the calibration curve (r = 0.9990 for PFOA, r = 0.9982 for PFOS) was obtained in the range of 0.05-5 ng mL⁻¹ each compound. The detection limits (S/N = 3) for PFOA and PFOS were 1.5 and 3.2 pg mL⁻¹, respectively. The method described here showed about 100-fold higher sensitivity than the direct injection method. The within-day and between-day precisions (relative standard deviations) were below 3.7 and 6.0%, respectively. This method was applied successfully to the analysis of PFOA and PFOS in environmental water samples and to the elution test from a Teflon^®-coated frying pan without interference peaks. The recoveries of PFOA and PFOS spiked into river samples were above 81%, and PFOA was detected at pg mL⁻¹ levels in environmental water samples and eluate from the frying pan.     

Temperature-response polymer coating for in-tube solid-phase microextraction coupled to high-performance liquid chromatography

The silica nanoparticle (SiO₂ NP)-deposited capillary fabricated by liquid phase deposition (LPD) was bonded by 3-(triethoxysilyl) propyl methacrylate and then modified with poly(N-isopropylacrylamide) (PNIPAAm) by polymerization. The resulting PNIPAAm modified SiO₂ NP-deposited capillary was applied to in-tube solid-phase microextraction coupled to high-performance liquid chromatography (in-tube SPME-HPLC). To investigate the extraction performance of the prepared capillary, diethylstilbestrol (DES) with moderate polarity was selected as the model analyte. Results demonstrate that PNIPAAm modified SiO₂ NP-deposited capillary exhibited obvious temperature responsive character. Finally, the PNIPAAm modified SiO₂ NP-deposited capillary was applied to the analysis of three synthetical estrogens from milk samples. The detection limit of the method was found to be in the range 1.2-2.2 ng/g, and recovery was 71.7-98.9% with relative standard deviations in the range of 2.8-12.6%.     

A critical review of vacuum-assisted headspace solid-phase microextraction for environmental analysis

Vacuum-assisted headspace solid-phase microextraction (Vac-HSSPME) is an emerging analytical technique, which further advances HSSPME by providing lower detection limits of analytes with poor volatility at shorter extraction times. This review discusses the theoretical aspects and possibilities of the Vac-HSSPME technique for analysis of environmental samples. Optimization of key parameters, currently available equipment and methods for quantification of organic pollutants in water and soil are considered. Key problems and limitations of the technique are discussed along with possible approaches for its future development. The technique has a well-developed theory, which could be used for modeling of the extraction process, faster method development, and optimization. Wider application of the technique is limited by the lack of automation, which, however, seems possible to develop and implement by manufacturers of commercial multi-purpose autosamplers for gas chromatography instruments. It has been shown that Vac-HSSPME allows decreasing cross-contamination of samples from the laboratory air, which is advantageous for identification and quantification of trace environmental pollutants. Simple equipment for the technique makes it possible to apply for on-site sample preparation and analysis of environmental samples.     

Determination of camptothecin and 10-hydroxycamptothecin in human plasma using polymer monolithic in-tube solid phase microextraction combined with high-performance liquid chromatography

A biocompatible in-tube solid-phase microextraction (SPME) device was used for the direct and on-line extraction of camptothecin and 10-hydroxycamptothecin in human plasma. Biocompatibility was achieved through the use of a poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column for extraction. Coupled to high performance liquid chromatography (HPLC) with UV detection, this on-line in-tube SPME method was successfully applied to the simultaneous determination of camptothecin and 10-hydroxycamptothecin in human plasma. The calculated detection limits for camptothecin and 10-hydroxycamptothecin were found to be 2.62 and 1.79 ng/mL, respectively. The method was linear over the range of 10–1000 ng/mL. Excellent method reproducibility was achieved, yielding RSDs of 2.49 and 1.59%, respectively. The detection limit (S/N=3) of camptothecin was found to reach 0.1 ng/mL using fluorescence detection. The proposed method was shown to cope robustly with the extraction and analysis of camptothecin and 10-hydroxycamptothecin in plasma samples.     

Determination of fluoroquinolones in environmental waters by in-tube solid-phase microextraction coupled with liquid chromatography-tandem mass spectrometry

We developed a sensitive and useful method for the determination of five fluoroquinolones (FQs), enoxacin, ofloxacin, ciprofloxacin, norfloxacin, and lomefloxacin in environmental waters, using a fully automated method consisting of in-tube solid-phase microextraction (SPME) coupled with liquid chromatography-tandem mass spectrometry (LC/MS/MS). These compounds were analysed within 7 min by high-performance liquid chromatography (HPLC) using a CAPCELL PAK C8 column and aqueous ammonium formate (pH 3.0, 5 mM)/acetonitrile (85/15, v/v) at a flow rate of 0.2 mL/min. Electrospray ionization conditions in the positive ion mode were optimized for MS/MS detection. In order to optimize the extraction of FQs, several in-tube SPME parameters were examined. The optimum in-tube SPME conditions were 20 draw/eject cycles of 40 μL of sample at a flow-rate of 150 μL/min, using a Carboxen 1010 PLOT capillary column as an extraction device. The extracted compounds were easily desorbed from the capillary by passage of the mobile phase. Using the in-tube SPME LC/MS/MS method, good linearity of the calibration curve (r ≥ 0.997) was obtained in the concentration range from 0.1 to 10 ng/mL for all compounds examined. The limits of detection (S/N = 3) of the five FQs ranged from 7 to 29 pg/mL. The in-tube SPME method showed 60-94-fold higher sensitivity than the direct injection method (5 μL injection). This method was applied successfully to the analysis of environmental water samples without any other pretreatment and interference peaks. Several surface waters and wastewaters were collected from the area around Asahi River, and ofloxacin was detected in wastewater samples of a sewage treatment plant and other two hospitals at 17.5-186.2 pg/mL. The recoveries of FQs spiked into river water were above 81% for a 0.1 or 0.2 ng/mL spiking concentration, and the relative standard deviations were below 1.9-8.6%.     

Hybrid organic-inorganic octyl monolithic column for in-tube solid-phase microextraction coupled to capillary high-performance liquid chromatography

An octyl-functionalized hybrid silica monolithic column was developed for in-tube solid-phase microextraction (SPME) to perform on-line preconcentration coupled to capillary high-performance liquid chromatography (microHPLC) analysis. A hybrid silica monolithic column functionalized with octyl groups was conveniently synthesized by a two-step acid/base-catalyzed hydrolysis/co-condensation of tetraethoxysilane (TEOS) and n-octyltriethoxysilane (C8-TEOS). The size of through-pores as well as the carbon content can be adjusted by changing the ratio of TEOS to C8-TEOS in the polymerization mixture. The extraction characteristics of the monolithic column prepared under optimized fabrication conditions were studied by using polycyclic aromatic hydrocarbons (PAHs) as the analytes. The sample volume that could be injected into the system was increased up to 1mL with simultaneous increase of column efficiency, when hybrid silica monolithic column was used as a precolumn. Good linear calibration curves (R>0.999) were obtained, and the limits of detection (signal-to-noise ratio, S/N=3) for the analytes were found to be between 2.4 and 8.1ng/mL with a UV absorbance detector, which are 299-456 times lower than those obtained without preconcentration. The column-to-column RSD values were 1.3-8.0% for recoveries of PAHs investigated.     

High-performance liquid chromatographic enantioseparations on capillary columns containing crosslinked polysaccharide phenylcarbamate derivatives attached to monolithic silica

Monolithic capillary columns containing native silica gel were covalently modified with 3,5-disubstituted phenylcarbamate derivatives of cellulose and amylose and applied for enantioseparations in capillary LC. The method previously used for covalent immobilization of polysaccharide phenylcarbamate derivatives onto the surface of microparticulate silica gel was successfully adapted for in situ modification of monolithic fused-silica capillary columns. The effects of the nature of polysaccharide and the substituents, as well as of multiple covalent immobilization of polysaccharide derivative on chromatographic performance of capillary columns were studied. The capillary columns obtained using this technique are stable in all solvents commonly used in LC and exhibit promising enantiomer resolving ability.