Depletion solid-phase microextraction for the evaluation of fiber-sample partition coefficients of pesticides

A depletion solid-phase microextraction (SPME) method for the characterisation of SPME sorption for 13 pesticides selected as probe compounds is described. The sample is extracted and analysed multiple times by SPME-GC/MS. The observed depletion in peak areas is used for the calculation of extraction ratios that varied between 3 and 28% for a PDMS fiber with confidence intervals between 0.7 and 5.4%. Apparent fiber-sample partition coefficients can be calculated and extrapolated to equilibrium conditions if specific sorption kinetics are known. Under the chosen conditions, problems were encountered for more polar compounds (logK(ow)<3) due to inefficient extraction. The extracted amount was found to be the decisive parameter for depletion SPME and the extraction conditions therefore need to be adapted to the polarity of the analyte. The importance of the initial analyte concentration especially for mixed-mode fibers is demonstrated. Compared with conventional external calibration using liquid injection, depletion SPME eliminates uncertainties due to solvent effects during injection. Furthermore, it does neither require authentic reference compounds nor knowledge of the initial analyte concentration, and thus can even be used for unknowns.     

Bio-compatible in-tube solid-phase microextraction capillary for the direct extraction and high-performance liquid chromatographic determination of drugs in human serum

A restricted access material (RAM), alkyl-diol-silica (ADS), was used to prepare a highly bio-compatible solid-phase microextraction (SPME) capillary for the automated and direct in-tube extraction of several benzodiazepines from human serum. The bifunctionality of the ADS extraction phase prevented fouling of the capillary by protein adsorption while simultaneously trapping the analytes in the hydrophobic porous interior. This the first report of a restricted access material utilized as an extraction phase for in-tube SPME. The approach simplified the required apparatus in comparison to existing RAM column switching procedures, and more importantly eliminated the excessive use of extraction solvents. The biocompatibility of the ADS material also overcame the existing problems with in-tube SPME that requires an ultrafiltration or other deproteinization step prior to handling biological samples, therefore further minimizing the sample preparation requirements. The calculated oxazepam, temazepam, nordazepam and diazepam detection limits were 26, 29, 22 and 24 ng/ml in serum, respectively. The method was linear over the range of 50-50 000 ng/ml with an average linear coefficient (R2) value of 0.9998. The injection repeatability and intra-assay precision of the method were evaluated with five injections of a 10-microg/ml serum sample (spiked with all compounds), resulting in an average RSD<7%. The ADS extraction column was robust, providing many direct injections of biological fluids for the extraction and subsequent determination of benzodiazepines.     

New high-performance cryofocalizer injector for in-tube solid-phase microextraction and headspace capillary gas chromatographic applications

The construction of a high-efficiency but cheap injector for volatile and very volatile compounds is shown. The device focuses the compounds in a fused-silica (FS) transfer capillary with the aid of liquid nitrogen. A 6.2 mm O.D. glass tube liner (ca. 25 cm x 1.5 mm I.D.) is inserted in the heated (-200 degrees C) injector of the gas chromatograph in place of the standard glass liner, and extends further externally through a liquid nitrogen container made with styrofoam-like material. Inside this glass tube, the FS transfer line passing through the oven door is connected like a pre-column to the analytical high-resolution GC column. It can move fast between the heated and the cooled zone (<-->, deltaL = 13 cm), and when this movement starts, crvofocused analytes are injected "at once" resulting in symmetrical and sharp injection bands with "zero" carryover. The performance of this device is demonstrated by its application to in-tube solid-phase microextraction and to spice volatiles analysis.     

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%.     

Monolithic silica column for in-tube solid-phase microextraction coupled to high-performance liquid chromatography

In-tube solid-phase microextraction (SPME) has successfully been coupled to capillary LC, and further an automated in-tube SPME system has been developed using a commercially available HPLC auto-sampler. However, an open tubular capillary column with a thick film of polymer (stationary phase) is unfavorable because the ratio of the surface area of coating layer contacted with sample solution to the volume of the capillary column is insufficient for mass transfer. A highly efficient SPME column is. therefore, required. We introduced a C18-bonded monolithic capillary column that was used for in-tube SPME. The column consisted of continuous porous silica having a double-pore structure. Both the through-pore and the meso-pore were optimized for in-tube SPME, and the optimized capillary column was connected to an HPLC injection valve for characterization. The results demonstrated that the pre-concentration efficiency is excellent compared with the conventional in-tube SPME. The novel method for both introduction and concentration of the samples was effective. satisfactory and suitable for use in the SPME medium.     

Comparison of solid-phase extraction and solid-phase microextraction for carbofuran in water analyzed by high-performance liquid chromatography-photodiode-array detection

In this study a direct solid-phase microextraction (SPME) procedure has been developed for the determination of carbofuran in water. Experimental parameters such as selection of SPME coating, effect of temperature, effect of salt addition and solvent desorption were studied and optimized. Analytical parameters such as linearity, precision, detection and quantitation limits, and matrix effects for solid-phase extraction (SPE) and SPME methods were evaluated for comparison purposes with the aim of selecting the most appropriate depending on the detection capabilities required. SPE and SPME were followed by high-performance liquid chromatography with diode-array detection, using a 50 x 4.6 mm I.D. guard column and a 150 x 4.6 mm I.D. analytical column, both packed with C18 silica. Both methods can be applied to real samples and give the same results, but SPE allows the detection of lower carbofuran concentrations (0.06 microg/L) as compared to     

Determination of liposome/water partition coefficients of organic acids and bases by solid-phase microextraction

The extraction of two methylated anilines and three chlorinated phenols by solid-phase microextraction (SPME) fibers coated with polyacrylate was investigated as a function of pH. Only the neutral species of the acids and bases partitioned into the polymer. Extraction kinetics were accelerated for the hydrophobic phenols at pH values around their acidity constant. This is presumably due to a reconstitution of the neutral species in the unstirred aqueous layer adjacent to the polymer surface by the charged species through the fast acid-base equilibrium. Although the charged species is not taken up into the polymer, liposome/water distribution ratios could be measured up to a pH value, where 99% of the compounds were present as charged species. The partition coefficients of the neutral and charged species were extrapolated from the pH profiles of the liposome/water distribution ratios. The resulting values were slightly lower than those measured with equilibrium dialysis. The discrepancies are discussed with respect to differences in the experimental conditions and the possibility of matrix effects during SPME measurements.     

Analysis of biogenic amines by solid-phase microextraction and high-performance liquid chromatography with electrochemical detection

We investigated the new solid-phase microextraction method by high-performance liquid chromatography with electrochemical detection for the analysis of biogenic amines. The Carbowax–Templated Resin 50 μm (purple) fibre coating offers good performances for dopamine and serotonin separation, i.e., good selectivity and high sensibility (0.1 μg l⁻¹). We also tested this fibre for biogenic amines quantification of rat striatum. The coating seems to be selective towards the amines and has low affinity for the metabolites, allowing a good separation and preventing drawbacks from the biological matrix. These first results obtained using this original separation method offer large perspectives of application to many biological samples.     

Validation of a solid-phase extraction high-performance liquid chromatographic assay for doxazosin.

The determination of doxazosin by high-performance liquid chromatography with fluorescence detection is described. Propanolol was used as the internal standard. Plasma samples were treated with methanol to precipitate the proteins. Doxazosin was isolated with C18 reversed-phase extraction columns. The determination limit is 1 ng/ml of plasma, while the extraction columns can be reused frequently. The method is applied to clinical trial samples.     

Strategies for interfacing solid-phase microextraction with liquid chromatography

Solid-phase microextraction (SPME) techniques are equally applicable to both volatile and non-volatile analytes, but the progress in applications to gas-phase separations has outpaced that of liquid-phase separations. The interfacing of SPME to gas chromatographic equipment has been straight-forward, requiring little modification of existing equipment. The requirement of solvent desorption for non-volatile or thermally labile analytes has, however, proven challenging for interfacing SPME with liquid-phase separations. Numerous options to achieve this have been described in the literature over the past decade, with applications in several different areas of analysis. To date, no single strategy or interface device design has proven optimal. During method development analysts must select the most appropriate interfacing technique among the options available. Out of these options three general strategies have emerged: (1) use of a manual injection interface tee; (2) in-tube SPME; and (3) off-line desorption followed by conventional liquid injection. In addition, there has been interest in coupling SPME directly to electrospray ionisation and matrix-assisted laser desorption ionisation (MALDI) for mass spectrometry. Several examples of each of these strategies are reviewed here, and an overview of their use and application is presented.     

Analysis of triazine in water samples by solid-phase microextraction coupled with high-performance liquid chromatography

Solid-phase microextraction (SPME) coupled with high-performance liquid chromatography (HPLC) for the determination of triazine is described. Carbowax/templated resin (CW/TPR, 50 μm), polydimethylsiloxane/divinylbenzene (PDMS/DVB, 60 μm), polydimethylsiloxane (PDMS, 100 μm), and polyacrylate (PA, 85 μm) fibers were evaluated for extraction of the triazines. CW/TPR and PDMS/DVB fibers were selected for further study. Several parameters of the extraction and desorption procedure were studied and optimized (such as types of fibers, desorption mode, desorption time, compositions of solvent for desorption, soaking periods and the flow rate during desorption period, extraction time, temperature, pH, and ionic strength of samples). Both CW/TPR and PDMS/DVB fibers are acceptable; a simple calibration-curve method based on simple aqueous standards can be used. The linearity of this method for analyzing standard solution has been investigated over the range 5-1000 ng mL⁻¹ for both PDMS/DVB and CW/TPR fibers. All the correlation coefficients in the range 5-1000 ng mL⁻¹ were better than 0.995 except Simazine and Atratone by CW/TPR fiber. The R.S.D.s range from 4.4% to 8.8 % (PDMS/DVB fiber) and from 2.4% to 7.2% (CW/TPR fiber). Method-detection limits (MDL) are in the range 1.2-2.6 and 2.8-3.4 ng mL⁻¹ for the two fibers. These methods were applied to the determination of trazines in environmental water samples (lake water).     

Determination of some aldehydes by using solid-phase microextraction and high-performance liquid chromatography with UV detection

A new approach has been developed for the extraction and determination of aldehydes such as veratraldehyde, m-nitrobenzaldehyde, cinnamaldehyde, benzaldehyde, and p-chlorobenzaldehyde by using solid-phase microextraction (SPME) and high-performance liquid chromatography with UV detection (HPLC/UV). The method involves adsorption of the aldehydes on polydimethylsiloxane/divinylbenzene-coated fiber, followed by desorption in the desorption chamber of the SPME-HPLC interface, using acetonitrile-water (70 + 30) as the mobile phase; UV detection was at 254 nm. A good separation of 5 aldehydes was obtained on a C18 column. The detection limits of veratraldehyde, m-nitrobenzaldehyde, cinnamaldehyde, benzaldehyde, and p-chlorobenzaldehyde are 25, 41, 13, 12, and 11 pg/mL, respectively, which are about 100 times better than the detection limits for other SPME methods using gas chromatography. The proposed method was validated by determining benzaldehyde in bitter almonds and cinnamaldehyde in cinnamon bark. The recoveries of the 5 analytes were determined by analysis of spiked drinking water.     

Development of an improved heated interface for coupling solid-phase microextraction to high-performance liquid chromatography

The aim of the study described in this report has been the development and the evaluation of a new improved interface to be operated under continuous heating, for on-line coupling solid-phase microextraction (SPME) to high-performance liquid chromatography (HPLC). Heating is desirable to increase desorption rate and decrease carryover. The results obtained have been compared with that obtained by off-line desorption and online desorption without heating. The SPME-HPLC interface described here has an inner volume of 60 microL, fixation for infinite points and a novel leak less sealing system. When the heating system was used, the area values were almost 10-fold higher than that obtained using the off-line mode. The obtained chromatograms showed an increasing of the area and height of chromatographic peaks and proved the excellent performance and reproducibility of the interface developed in this work.     

In-tube solid-phase microextraction with poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary for direct high-performance liquid chromatographic determination of ketamine in urine samples

Ketamine was used for anaesthesia originally but has emerged as an abused drug in recent years. The prevalence of ketamine abuse demands a direct and rapid determination method. It is known that in-tube solid phase microextraction (in-tube SPME) can perform extraction with a capillary linked directly to a HPLC system, providing an automated and accurate extraction procedure. In this paper, an in-tube SPME coupled to HPLC method was developed for the determination of ketamine in urine samples with a poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column as the extraction phase, which is expected to provide higher extraction efficiency than open tubular capillaries. After optimizing the extraction conditions, ketamine was extracted directly from urine samples in a wide dynamic linear range of 50-10,000 ng mL(-1), with the detection limit obtained as 6.4 ng mL(-1). The intra-day and inter-day precision for the method was 1.6% and 1.7%, respectively. The urine samples from suspect addicts have been successfully analyzed within 20 min. The re-usability of the monolithic column was also confirmed as no decrease of the extraction efficiency was shown after urine extraction.     

Rapid headspace gas chromatographic method for the determination of liquid/gas partition coefficients

A rapid, efficient and low-cost headspace technique useful for the determination of liquid/gas partition coefficients of gases and volatile substances of low and intermediate solubility is described. The equilibration step is carried out at constant pressure using glass syringes, with a ratio of liquid/gas phase volumes of ca. 1:3; after 30 min at the desired temperature, the headspace is recovered by transfer into another syringe and analyzed by gas chromatography. A study of the partition coefficients in water at 37 degrees C of 27 volatile compounds demonstrated that the method is fully applicable for all gases, with exception of those with a partition coefficient higher than 300.     

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%.     

Two high-performance liquid chromatographic methods for quantitation of theophylline in plasma

Two high-performance liquid chromatographic methods are described for the assay of theophylline in plasma. Both allowed the separation of theophylline from the caffeine metabolites, theobromine and 1,7-dimethylxanthine. Method A, using 8-chlorotheophylline as internal standard, involved back extraction of theophylline from organic extract with 0.1 M sodium hydroxide. Method B used generally accepted solvent extraction followed by evaporation and beta-hydroxyethyltheophylline as internal standard. High-performance liquid chromatographic analyses were performed on reversed-phase phenyl columns (25 X 0.46 and 25 X 0.41 cm) using 20% methanol in 20 mM phosphate buffer at pH 5.6 for Method A and 2% acetonitrile and 8% methanol in 20 mM phosphate buffer for Method B. The column effluent was monitored at UV 273 nm. Standard curves for both Methods A and B were fitted by linear regression (r greater than 0.999) in the concentration range of 0.05-50 micrograms/ml. Either method was selective, accurate and reproducible over the concentration range 0.08-26 micrograms/ml. However, compared with Method B, Method A provided significant advantages in terms of simplicity, speed and efficiency.     

Thin-layer chromatographic scanner, spectrophotometric and high-performance liquid chromatographic methods for the determination of colchicine

One high-performance liquid chromatographic (HPLC) and two thin-layer chromatographic (TLC) methods are proposed for the determination of colchicine in crude drugs and pharmaceutical preparations. The TLC scanner method is based on measurement of the absorbance of the separated colchicine spot; alternatively, after scraping the spot from the plate and elution the absorbance can be measured spectrophotometrically. The HPLC assay was carried out isocratically on a reversed-phase column using MeOH-H2O (60 + 40). The recoveries were 99.2 +/- 1.23, 99.1 +/- 1.12 and 99.1 +/- 2.01% for the TLC scanner, spectrophotometric and HPLC methods, respectively. The methods were shown to be sensitive and specific and can be used as an alternative to the pharmacopoeial methods having been applied to the determination of colchicine in corms of Merendera persica and in three pharmaceutical preparations.     

New materials for solid-phase extraction and multiclass high-performance liquid chromatographic analysis of pesticides in grapes

Sample preparation procedures which included the use of new aminopropyl (NH2) and octadecyl (C18) solid-phase extraction (SPE) sorbents are proposed for the simultaneous multiclass determination of the fungicide benomyl and of the herbicides tebuthiuron, diuron, simazine, atrazine, and ametryn in grapes, using single wavelength high-performance liquid chromatography. Sorbent preparation uses a fast, easy, and effective procedure to obtain silica-based materials, made by depositing polysiloxanes on a silica support followed by thermal immobilization. Recovery results of the compounds, after elution from the SPE cartridges, indicate that the most efficient system employed silica loaded with 40% of an aminofunctional polydimethylsiloxane as sorbent, using dichloromethane:methanol (95:5, v/v) as eluent. Method validation, carried out in agreement with International Conference on Harmonization directives, was performed at three fortification levels (100, 200, and 1000 microg kg(-1)). Limits of detection and quantification show that the method developed can be used to detect the pesticides at concentrations below the maximum residue levels established by Codex Alimentarius, the US Environmental Protection Agency, the European Union, and Brazilian legislation.     

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.