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We examine the potential of ultra-high-performance supercritical fluid chromatography for multiresidue quantification of ten pesticides commonly applied to lettuce and compares it to ultra-high-performance liquid chromatography. Initially, a thorough study of the stationary and mobile phase composition and injection solvent was carried out. In a second step, a chemometric approach based on design of experiments was used to simultaneously study the influence of temperature, pressure, and percentage of ethanol on the retention, resolution and symmetry of the peaks. Using this approach, it was possible to obtain the Design Space, a robust region where complete separation of the analytes was achieved, with acceptable peak shape. Both methods were validated according to the figures of merit: selectivity, linearity, quantification limit, accuracy (in terms of recovery), and precision (repeatability and intermediate precision) and used to quantify the pesticides in lettuce samples. Comparing both techniques, it was concluded that the limits of quantification, accuracy, and precision were similar. However, in supercritical fluid chromatography, a reduced volume of organic solvent was used, the method was faster and generated lower amounts of residues., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A new analytical method for multiresidue determination of 16 multiclass pesticides in lettuce was developed using ultra-high performance liquid chromatography with tandem mass spectrometry with a triple quadrupole mass analyzer and positive mode electrospray ionization, using a previously optimized quick, easy, cheap, effective, rugged, and safe method for sample preparation. Validation studies, according to document SANTE/11945/2015, demonstrated that the developed method is selective, accurate, and precise, providing recoveries of 70-120%, relative standard deviations ≤20% and quantification limits from 3 μg/kg. The method was compared with one based on high-performance liquid chromatography with tandem mass spectrometry, in terms of chromatographic performance, detectability and matrix effect for five varieties of lettuce. The new method provided a reduction in the time for the chromatographic analysis of 50%, from 30 to 15 min, using a lower mobile phase flow rate (0.147 mL/min), which reduced the consumption of mobile phase by 25%, and injection of smaller amounts of sample (1.7 μL). Lower limits of quantification were obtained for almost all pesticides studied for green-leaf lettuce. However, in relation to the matrix effect, four of the five types of lettuce studied presented higher matrix effects., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A new analytical method using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) procedure for multi-residue determination of 16 multiclass pesticides in five different types of lettuce was developed using high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry. The QuEChERS procedure was optimized in terms of extractor solvent, partitioning salts and clean-up salts, through recovery, gravimetric analysis and matrix-effect studies. Microwave extraction and the use of disposable pipette extraction in the clean-up step were also tested, providing interesting alternatives to the traditional QuEChERS method, depending on the pesticides properties. The use of a laboratory-made phenyl-type sorbent, based on silica and poly(2-phenylpropyl)methylsiloxane, was studied in the clean-up step, presenting promising results as a substitute for primary secondary amine (PSA). The optimized QuEChERS method was defined employing acetonitrile, citrate buffer and clean-up with PSA, MgSO 4 and graphitized carbon black. The method developed was validated according to Document SANTE/11945/2015 and proved to be selective, accurate and precise, obtaining limits of quantification from 5μgkg -1 and recoveries in the range of 70-120% with relative standard deviation≤20%. The method was applied on 14 real samples from commercial markets in Brazil and 21% of the samples analyzed presented irregularities, according to local pesticides regulations., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A new molecularly imprinted polymer modified with restricted access material (a hydrophilic external layer), (MIP-RAM) was synthesized via polymerization in situ in an open fused silica capillary. This stationary phase was used as sorbent for in-tube solid phase microextraction (in-tube SPME) to determine parabens in breast milk samples by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Scanning electron micrographs (SEM) illustrate MIP surface modification after glycerol dimethacrylate (hydrophilic monomer) incorporation. The interaction between parabens and MIP-RAM was investigated by Fourier-transform infrared (FTIR) spectroscopy. The Scatchard plot for MIP-RAM presented two linear parts with different slopes, illustrating binding sites with high- and low-affinity. Endogenous compounds exclusion from the MIP-RAM capillary was demonstrated by in-tube SPME/LC-UV assays carried out with blank milk samples. The in-tube SPME/UHPLC-MS/MS method presented linear range from 10 ng mL(-1) (LLOQ) to 400 ng mL(-1) with coefficients of determination higher than 0.99, inter-assay precision with coefficient of variation (CV) values ranging from 2 to 15%, and inter-assay accuracy with relative standard deviation (RSD) values ranging from -1% to 19%. Analytical validation parameters attested that in-tube SPME/UHPLC-MS/MS is an appropriate method to determine parabens in human milk samples to assess human exposure to these compounds. Analysis of breast milk samples from lactating women demonstrated that the proposed method is effective., (Copyright © 2016 Elsevier B.V. All rights reserved.)

A new reversed phase high-performance liquid chromatography (RP-HPLC) stationary phase was prepared and its chromatographic and physical-chemical properties were evaluated. The new stationary phase was prepared with a silica support and poly(2-phenylpropyl)methylsiloxane (PPPMS), a phenyl type polysiloxane copolymer. Since this is a new copolymer and there is little information in the literature, it was submitted to physical-chemical characterization by infrared spectroscopy and thermogravimetry. The chromatographic phase was prepared through sorption and microwave immobilization of the copolymer onto a silica support. The chromatographic performance was evaluated by employing test procedures suggested by Engelhardt and Jungheim, Tanaka and co-workers, Neue, and Szabó and Csató. These test mixtures provide information about the hydrophobic selectivity, silanophilic activity, ion-exchange capacity, shape selectivity and interaction with polar analytes of the new Si-PPPMS reversed phase. Stability tests were developed using accelerated aging tests under both basic and acidic conditions to provide information about the lifetime of the packed columns., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Independent methods for determination of organic contaminants such as pharmaceuticals and pesticides in drinking water samples, using SPE as the extraction technique and LC-MS/MS in the MRM mode with electrospray ionization, were developed and validated. Different SPE sorbents were evaluated, including lab-made fluorinated and phenyl and commercial Oasis HLB and C18, with the commercial phases being more suitable for the target compounds. Recoveries in the range of 70-120% were obtained for all target compounds, with the exception for paracetamol (acetaminophen), and precision values (inter-day and intra-day), expressed in terms of relative standard deviations (RSD), lower than 20% were obtained for all target compounds. Quantification limits were in the range of 0.006-0.208 μg L(-1) and the methods developed were successfully applied for the analysis of drinking water samples, detecting some pharmaceuticals and pesticides, but at concentration levels lower than the MRL., (Copyright © 2012 Elsevier B.V. All rights reserved.)

A procedure involving hollow fiber microporous membrane liquid-liquid extraction (HF-MMLLE) and detection by liquid chromatography with tandem mass spectrometry was developed and applied. The extraction is based on liquid-liquid microextraction with a polypropylene porous membrane as a solid support for the solvent. On the membrane walls the solvent forms a renewable liquid membrane which improves the trueness of the method and promotes the sample clean-up. The applicability of this method was evaluated through the simultaneous extraction of 18 pesticides of different classes: polar organophosphates, carbamates, neonicotinoids, amides, pyrimidines, benzimidazoles and triazoles in industrial and fresh orange juice. The parameters affecting the extraction efficiency were optimized by multivariable designs. Under optimized conditions, analytes were concentrated onto 1.5 cm long microporous membranes placed directly into the sample containing 9mL of juice at pH 7.0, 4 g of ammonium sulfate and 400 μL of toluene:ethyl acetate (85:15, v/v). The best extraction conditions were achieved at 25°C with 35 min of extraction time. The analyte desorption was carried out using 50 μL of methanol:acetone (50:50, v/v) for 2 min in an ultrasonic bath. Limits of detection ranging between 0.003-0.33 mgL(-1), 0.003-0.35 mg L(-1) and 0.003-0.15 mgL(-1) were obtained for the carton orange juice, carton light orange juice and fresh orange juice samples, respectively. Good repeatability (lower than 7.6%) was obtained for all three sample types. The method was applied to five different juice samples containing soybean extract, orange pulp, nectar, light juice and fresh orange juice. The results suggest that the proposed method represents a very simple and low-cost alternative microextraction procedure rendering adequate limits of quantification for the determination of these pesticides in juice samples., (Copyright © 2011 Elsevier B.V. All rights reserved.)

A sensitive and precise analysis using hollow fiber renewal liquid membrane (HFRLM) extraction followed by high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) is described for determination of five sulfonamides in honey samples. In this procedure, the organic solvent introduced directly into the sample matrix extracts the sulfonamides and carries them over the polypropylene porous membrane. An organic solvent is immobilized inside the polypropylene porous membrane, leading to a homogeneous phase. The stripping phase at higher pH in the lumen of the membrane promotes the ionization of the target compounds releasing them to this phase. The most important parameters affecting the extraction efficiency were optimized by multivariable designs (pH and sample mass, pH and buffer for stripping phase, extraction temperature and time, type and volume of extractor solvent and use of salt to saturate the sample). Detection limits in the range of 5.1-27.4 μg kg(-1) and linearity coefficient of correlation higher than 0.987 were obtained for the target analytes. The results obtained for the proposed method show that HFRLM-LC-MS/MS can be used for determination of the five sulfonamides studied in honey samples with excellent precision, accuracy, practicality and short analysis time., (Copyright © 2010 Elsevier B.V. All rights reserved.)

At the present time, more complex analyses of apolar compounds with similar chemical structures or of polar compounds, especially basic ones, having diverse properties require more selective stationary phases having better stabilities. This paper describes several new stationary phases with directed selectivities that were prepared by immobilizing two different phenyl group-containing siloxanes and a trifluoropropyl-containing siloxane onto chromatographic silica and, in the case of the fluorinated siloxane, onto zirconized silica, using thermal treatment or microware radiation. The chromatographic properties and stabilities of these new phases were determined and several applications were evaluated. The phenyl-containing phases showed excellent characteristics related to the separation of several different types of aromatic compounds while the fluorinated phases, which present a more polar character, revealed selectivity for the separation of positional isomers as well as for a mixture of basic pharmaceuticals. Stability tests indicate that immobilization of the polysiloxanes increases column lifetimes by making the stationary phases less susceptible to dissolution, while the phases immobilized with microwave radiation were somewhat more stable than those immobilized by thermal treatments.

A method for the determination of artemether (ART) and its main metabolite dihydroartemisinin (DHA) in plasma employing liquid-phase microextraction (LPME) for sample preparation prior to liquid chromatography-tandem mass spectrometry (LC-MS-MS) was developed. The analytes were extracted from 1mL of plasma utilizing a two-phase LPME procedure with artemisinin as internal standard. Using the optimized LPME conditions, mean absolute recovery rates of 25 and 32% for DHA and ART, respectively, were achieved using toluene-n-octanol (1:1, v/v) as organic phase with an extraction time of 30min. After extraction, the analytes were resolved within 5min using a mobile phase consisting of methanol-ammonium acetate (10mmolL(-1), pH 5.0, 80:20, v/v) on a laboratory-made column based on poly(methyltetradecylsiloxane) attached to a zirconized-silica support. MS-MS detection was employed using an electrospray interface in the positive ion mode. The method developed was linear over the range of 5-1000ngmL(-1) for both analytes. Precision and accuracy were within acceptable levels of confidence (<15%). The assay was applied to the determination of these analytes in plasma from rats treated with ART. The two-phase LPME procedure is affordable and the solvent consumption was very low compared to the traditional methods of sample preparation.

A new fluorinated stationary phase was prepared through thermal immobilization of poly(methyl-3,3,3-trifluoropropylsiloxane) onto 5 microm Kromasil silica particles. The best conditions of immobilization time and temperature were determined through a central composite design and response surface methodologies. Physical-chemical characterization using solid-state (29)Si NMR measurements, infrared spectroscopy and elemental analysis showed that the immobilization process was effective to promote a coating of the support that corresponds to a monolayer of polymer. The stationary phase presents selectivity for positional isomers and good peak shape for basic compounds.

Many chromatographic separations require high pH mobile phases for the elution of basic compounds with symmetrical peaks appropriate for quantitation. Phosphate and carbonate salts are the most common additives used to achieve these conditions. However, they act in different ways on the stability of silica-based stationary phases. To provide a faster evaluation of the effects of these high pH solutions on the column lifetime of new immobilized-polymer metalized-silica-based phases prepared in our laboratory, an accelerated stability test at a nominal pH of 10 and at 50 degrees C was developed and applied with monitoring of the retention factor, column efficiency, and asymmetry as functions of the volume of high pH mobile phase. The laboratory-made stationary phases presented chemical stabilities in phosphate solution more than three times better their chemical stabilities in carbonate buffer.

A statistical approach for the simultaneous optimization of the mobile and stationary phases used in reversed-phase liquid chromatography is presented. Mixture designs using aqueous mixtures of acetonitrile (ACN), methanol (MeOH) and tetrahydrofuran (THF) organic modifiers were performed simultaneously with column type optimization, according to a split-plot design, to achieve the best separation of compounds in two sample sets: one containing 10 neutral compounds with similar retention factors and another containing 11 pesticides. Combined models were obtained by multiplying a linear model for column type, C8 or C18, by quadratic or special cubic mixture models. Instead of using an objective response function, combined models were built for elementary chromatographic criteria (retention factors, resolution and relative retention) of each solute or pair of solutes and, after their validation, the global separation was accomplished by means of Derringer's desirability functions. For neutral compounds a 37:12:8:43 (v/v/v/v) percentage mixture of ACN:MeOH:THF:H2O with the C18 column and for pesticides a 15:15:70 (v/v/v) ACN:THF:H2O mixture with the C8 column provide excellent resolution of all peaks.

An endcapped stationary phase is prepared by thermal immobilization of poly(methyltetradecylsiloxane) (PMTDS) onto a doubly zirconized silica support followed by endcapping using a mixture of hexamethyldisilazane and trimethylchlorosilane. The preparation of the Si-Zr(PMTDS)ec phase shows good repeatability with RSD <3.0% for carbon loadings and column efficiency. This new stationary phase has a lower density of residual hydroxyl groups, according to spectroscopic methods while basic compounds from the Tanaka and Engelhardt test mixtures are eluted with essentially symmetric peaks. Furthermore, the stability of the Si-Zr(PMTDS)ec stationary phase, measured using an accelerated aging test, is twice as great as the stability of a similar nonendcapped phase. The new phase shows promise for the separation of basic pharmaceuticals.

This paper describes fundamentals and applications of multivariate statistical techniques for the optimization of chromatographic systems. The surface response methodologies: central composite design, Doehlert matrix and Box-Behnken design are discussed and applications of these techniques for optimization of sample preparation steps (extractions) and determination of experimental conditions for chromatographic separations are presented. The use of mixture design for optimization of mobile phases is also related. An optimization example involving a real separation process is exhaustively described. A discussion about model validation is presented. Some applications of other multivariate techniques for optimization of chromatographic methods are also summarized.

The models parallel factor analysis (PARAFAC) and the recently introduced bilinear least squares (BLLS) were applied to develop second-order calibration methods to high performance liquid chromatography with diode array detection (HPLC-DAD) data, where overlap of interferences with the compounds of interest was observed, making the determination and resolution of the analytes possible. In this work, the simultaneous determination of five pesticides and two metabolites in wine samples by HPLC-DAD was performed, using the second-order advantage. The results of two chromatographic methods were compared, involving either isocratic or gradient elution. An appropriate preprocessing method was necessary to correct the effects of time shifts, baseline variations and background. BLLS presented results that were of the same quality as PARAFAC in five cases, but in two other situations only PARAFAC enabled analyte quantitation. Relative errors of prediction lower than 10% for all compounds were obtained, indicating that the methodology employing HLPC-DAD and second-order calibration can handle complex analytical systems.

A new material based on poly(methyltetradecylsiloxane) (PMTDS) thermally immobilized onto a silica support has been tested as a sorbent for the solid-phase extraction (SPE) from water of several pesticides used in soybean cultivation. The SPE methodology was developed and validated for six of these pesticides (imazethapyr, imazaquin, metsulfuron-methyl, bentazone, chlorimuron-ethyl and tebuconazole) according to the International Conference on Harmonization directives and the results were compared with those obtained with a commercial C18 SPE cartridge. The PMTDS-based sorbent gives results similar to the commercial sorbent with recoveries and precisions in agreement with directives for residue analysis. The quantification limits, after concentration, of all the pesticides evaluated were 1.0 microg L(-1), below the levels imposed by the principal regulatory agencies. The PMTDS-based sorbent preparation is fast, easy and reproducible and the cartridges are less expensive than similar commercial SPE materials.

Near infrared emission spectroscopy (NIRES) has been investigated in the study of different materials employed in liquid chromatography. The samples were heated in a nitrogen atmosphere and the emission spectra were obtained using a lab-made NIRES instrument. Through principal component analysis (PCA) using the raw emission spectra, it was possible to distinguish different materials according to their physical and/or chemical characteristics. Linear relationships between emissivity spectra and the contents of the coating material or the specific surface areas was observed for stationary phases or bare silicas, respectively. Furthermore, the thermal stability of stationary phases could be followed in real time.

A laboratory-made sorbent for solid-phase extraction (SPE) was obtained by thermal immobilization of poly(methyloctylsiloxane) (PMOS) onto silica. Cartridges packed with the new sorbent were used for the simultaneous determination of imazethapyr, nicosulfuron, diuron, linuron and chlorimuron-ethyl in water. These pesticides were separated and quantified using high-performance liquid chromatography with diode array detection (HPLC-DAD). The recoveries achieved with the laboratory-made PMOS cartridges were compared with those of some commercially available silica-based and polymer-based cartridges having C18, C8 and NH(2) pendant groups. Method validation using the laboratory-made sorbent was performed for the five pesticides at three fortifications levels (1x, 2x and 10x the limit of quantification of each pesticide). The laboratory-made PMOS cartridge has low cost preparation and showed good recoveries (72-111%) for all pesticides. Repeatability and intermediate precision were lower than 15%. Its performance was similar or even better, in some cases, than those of the commercial cartridges.

Immobilized presynthesized polymers on porous metalized (zirconized or titanized) silica particles as new stationary phases with improved chemical stability for RP-HPLC are reviewed. The preparations using different polymers, such as poly(methyloctylsiloxane), poly(methyltetradecylsiloxane), and poly(butadiene), different immobilization steps (gamma radiation, thermal treatment, and microwave radiation), and the chromatographic performances of these phases for polar, apolar, acidic, and basic compounds are discussed. The stability of some of these stationary phases using alkaline mobile phases is also presented.

Titanium oxide-modified silica was prepared by reaction of silica with titanium tetrabutoxide and then was used as support in the preparation of stationary phases with self-immobilized polybutadiene (PBD) and PBD immobilized through microwave radiation. Chromatographic performance of the stationary phases was evaluated in terms of the efficiency (plates/m), asymmetry (A(s)), retention factor (k) and resolution (R(s)) of two standard sample mixtures, one of then containing the basic compound N,N-dimethylaniline. A microwave irradiation of 30 min at 520 W gave the best efficiency (86,500 N m(-1)), greater than that of a 6-day self immobilized phase (69,500 N m(-1)). Self-immobilized stationary phases prepared with bare silica were also studied for comparison. These resulted in lower chromatographic performance, 43,800 N m(-1), when compared to the self-immobilized phase prepared with titanized silica.

A method for high-performance liquid chromatographic (HPLC) multiresidue determination of six pesticides in tomatoes was developed and validated. Silica-based C18 (octadecyl) and NH2 (aminopropyl) solid-phase extraction (SPE) sorbents, made in our laboratory, were used for sample preparation. The SPE materials were obtained by thermal immobilization of appropriate polysiloxanes onto 40 microm silica surfaces and were used in sample preparation for multiresidue analysis of the following pesticides: tebuthiuron and diuron (urea herbicides), simazine, atrazine and ametryn (triazines herbicides) and benomyl (benzimidazol fungicide). The results were compared with similar commercial materials. Reversed-phase high-performance liquid chromatography (RP-HPLC) using a Purospher RP-18 5 microm column and ACN: 0.01% aqueous NH4OH, pH 8.4 (35:65, v/v) as mobile phase, at 0.7 mlmin(-1), with 235 nm UV detection, was used for separation and quantification of the pesticides. Method validation was performed at three fortification levels (100, 200, 1000 microg l(-1)). Limits of detection and quantification show that the methods developed can be used to detect the pesticides in concentrations below the maximum residue levels (MRL) established by the Codex Alimentarius, USA, European Union and Brazilian legislations. The results showed that aminopropyl materials have a better performance than the octadecyl sorbents. Laboratory-made materials give results similar to commercial sorbents, with recoveries and precisions in agreement with directives for method validation in residue analysis.

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.

Polybutadiene (PBD) has been immobilized on high-performance liquid chromatography (HPLC) silica by microwave radiation at various power levels (52-663 W) and actuation times (3-60 min). Columns prepared from these reversed-phase HPLC materials, as well as from similar non-irradiated materials, were tested with standard sample mixtures and characterized by elemental analysis (%C) and infrared spectroscopy. A microwave irradiation of 20 min at 663 W gives a layer of immobilized PBD that presented good performance. Longer irradiation times give thicker immobilized layers having less favorable chromatographic properties.

A 100 ml bottle of polybutadiene, PBD, was repeatedly exposed to air over a period of 6 months. Samples were taken at time zero (PBD-0), after 3 months (PBD-3) and 6 months (PBD-6). These samples were sorbed onto HPLC silica by an open-air solution-evaporation procedure, which involved exposure to the atmosphere for 6 days. Portions of the three sets of samples were used to compare self-immobilization and the effects of 100 degrees C thermal treatments in air or nitrogen on HPLC performance of the resulting phases. It is concluded that self-immobilization is enhanced by prior exposure of sorbed PBD to air and subsequent heating at 100 degrees C further enhances column performance. The best performance (10(5) plates m(-1)) resulted from 4 h heating of PBD-6 material in nitrogen.

Chromatographic evaluations of a C18 dimethylurea phase in 150 mm x 3.9 mm HPLC columns were performed using the Tanaka and Engelhardt test mixtures. The applicability of the new C18 dimethylurea phase was also evaluated with a mixture of some herbicides and their metabolites. An artificial aging procedure was also performed by passing a potassium phosphate mobile phase buffered at pH 7.0 through C18 50 mm x 3.9 mm dimethylurea columns. The column stability was evaluated by means of the chromatographic parameters obtained for the separation of some compounds from the Neue test mixture, using apolar, polar and highly basic analytes.

A method for the simultaneous determination of diuron and linuron pesticides in human urine was developed, using both solid-phase extraction (SPE) and high-performance liquid chromatography (HPLC) phases made in our own laboratory. These materials were prepared by sorption of polysiloxanes onto a silica surface, followed by immobilization. The HPLC columns were prepared from poly(methyloctylsiloxane), PMOS, immobilized onto silica with microwave radiation while the SPE cartridges where made with poly(methyloctadecylsiloxane), immobilized thermally. Method validation was performed for diuron and linuron for three fortification levels. The recoveries obtained were 85-103%, the inter- and intra-assay precisions were less than 1.6 and 1.8%, respectively. The limits of quantitation and detection for diuron were 2.4 and 8.0 microg/l and for linuron were 5.0 and 12 microg/l, respectively.

Polybutadiene (PBD) has been immobilized on HPLC silica by gamma radiation doses in the range from 5 to 180 kGy. Columns prepared from these reversed-phase materials, as well as from similar non-irradiated materials, were tested with standard sample mixtures and characterized by elemental analysis (% C) and infrared spectroscopy. A low dose of 5 kGy is sufficient to produce a layer of immobilized PBD which functions as an efficient and stable stationary phase. Higher doses give thicker immobilized layers having less favorable chromatographic properties.

A method was developed to determine simazine, atrazine and their metabolite, 2-chloro-4,6-diamino-1,3,5-triazine, in urine. The presence of these herbicides in urine may reflect possible exposure to pesticides. Sample preparation involved protein precipitation and solid-phase extraction. The samples were analyzed by high-performance liquid chromatography-mass spectrometry. The detection limits were 0.4 microg/l and the analytes have a linear response in the interval 6-800 microg/l. The precision of the method was reflected in the RSD of < 2.4% for the herbicides studied. Based on the detectable herbicide levels from spiked urine samples collected from unexposed volunteers, this method can be used to determine the low levels necessary for establishing reference values of the selected herbicides and the metabolite.

New monofunctional C18 urea stationary phases with sterically protecting dimethyl and diisopropyl groups were prepared by a single step modification process. ProntoSil spherical silica (3 microm) was chemically modified with the monofunctional ethoxysilanes, [(3-octadecylurea)propyl]dimethyl and [(3-octadecylurea)propyl]diisopropyl ethoxysilanes. The phases were characterized by elemental analysis, infrared and solid-state 29Si and 13C NMR spectroscopies. Chromatographic characterizations of the new phases in 50x3.9 mm HPLC columns were performed by the separation of two different test mixtures, containing nonpolar, polar and highly basic compounds.

Chromatographic evaluations of a new C18 urea phase in 150x3.9 mm HPLC columns, involving the separation of different test mixtures, indicate good performance for both polar and basic compounds when compared with a commercial C18 reversed phase and also show promising results for the separation of some herbicides. An aging study was performed by passing a potassium phosphate mobile phase buffered at pH 7 through 50x3.9 mm HPLC columns. The column stability was evaluated by means of the chromatographic parameters obtained for the separation of some compounds of the Neue test mixture, containing apolar, polar and highly basic analytes. The applicability of the new C18 urea phase was evaluated with a herbicide mixture.

Five different reversed-phase materials for high-performance liquid chromatography were obtained by deposition of poly(methyloctylsiloxane) in HPLC silica particles, followed by immobilization using different processes: thermal treatment (120 or 220 degrees C for 4 h), irradiation with microwaves (495 W for 15 min), gamma radiation (dose of 80 kGy) and self-immobilization. These phases were characterized by gel permeation chromatography, percent carbon, 13C and 29Si solid-state nuclear magnetic resonance spectroscopy, infrared spectroscopy, thermogravimetric analysis and scanning electron microscopy. The results show that the different immobilization processes produce different physical characteristics in the prepared phases.

A new C18 urea stationary phase was prepared by a single-step modification process through the reaction of ProntoSil spherical silica (3 microm, Bischoff) with the trifunctional alkoxysilane (CH3CH2O)3-Si-(CH2)3-NH-C(O)-NH-(CH2)17-CH3, prepared in our laboratory. The phase was characterized by elemental analysis and solid-state 29Si and 13C nuclear mangnetic resonance spectrometry. Chromatographic evaluations of the new phase in 150 x 3.9 mm HPLC columns involving the separation of different test mixtures, indicate good performance for both polar and basic mixtures and also show promising results for further applications.

This work presents a fast, easy and effective procedure to obtain silica-based C18 type sorbents for use in solid-phase extraction (SPE). The solid-phase was made by depositing a polymer, poly(methyloctadecylsiloxane) (PMODS), on the silica support followed by immobilization using gamma irradiation. The method presents an application of green chemistry in the analytical laboratory, since there are no toxic residues after the synthesis. Water samples spiked with a mixture of pesticides (benomyl, tebuthiuron, diuron, simazine, atrazine and ametryn) were extracted in the off-line mode in order to evaluate the performance of this sorbent.

New urea-functionalized silica stationary phases were prepared by a single-step surface modification through reaction of LiChrosorb Si100 (5 microm particle size) with a homologous series of alkoxysilanes, synthesized in our laboratory, with the general formula (CH3CH2O)3Si(CH2)3NHC(O)NH(CH2)nCH3, where n=4, 6 and 11. The modified silicas were characterized by elemental analysis of carbon and nitrogen, solid-state 29Si- and 13C-cross polarization magic angle spinning nuclear magnetic resonance and nitrogen adsorption isotherms at 77 K. Chromatographic evaluation of the three urea-functionalized silicas in 150x3.9 mm I.D. HPLC columns was carried out by the separation of a test mixture composed of uracil, acetophenone, benzene, toluene and naphthalene, using acetonitrile-water as mobile phase. These new stationary phases, with embedded polar urea groups, are very promising when compared with amide phases prepared by the conventional two-step modification process. A single-step reaction process silica modification is better for obtaining a well-characterized and homogeneous modified surface.

Reversed-phase packing materials were prepared from HPLC silica and from zirconized HPLC silica support particles having sorbed poly(methyloctylsiloxane) (PMOS) as the stationary phase. Portions of zirconized material were subjected to 80 kGy of ionizing radiation. Columns prepared from these packing materials were subjected to 5000 column volumes each of neutral and alkaline (pH 10) mobile phases, with periodic tests to evaluate chromatographic performance. It was shown that the PMOS stationary phase sorbed onto zirconized silica requires an immobilization treatment (such as gamma irradiation) for long term stability while prior surface zirconization of the silica support surface greatly improves the chromatographic stability of the stationary phase when using alkaline mobile phases.

A method for determination of some triazine residues in water has been developed. The method involves concentration with C18 solid-phase extraction cartridges followed by high-performance liquid chromatographic analysis using a C18 column with UV detection at 230 nm, a mobile phase of methanol-water (60:40, v/v) at pH 4.6 (phosphoric acid) and a flow-rate of 0.8 ml/min. After optimization of the extraction and separation conditions, the method was validated. The method can be used for determination of atrazine, simazine, cyanazine and ametryn in water, within the international limits of 0.1 microg/l.

Poly(methyloctylsiloxane) (PMOS) was sorbed into the pores of HPLC silica by a solvent evaporation procedure, then irradiated with gamma rays from a cobalt-60 source to absorbed doses in the range from 0 to 200 kGy (1Gy = 1J kg-1). Non-irradiated and irradiated samples were characterized by solvent extraction, specific surface area determination, infrared spectroscopy and reversed-phase column performance. Solvent extraction data reveal that about 40% of the PMOS is not extractable prior to irradiation and this increases to about 75% with radiation doses of 50 kGy or higher. Column performance was improved by the radiation treatment, reaching a maximum efficiency in the dose range of 80-140 kGy while the peak symmetry changed from As = 1.7 to 1.1. The improvement is attributed to the increased mass of polymer immobilized by the radiation treatment and to a more uniform distribution of the immobilized polymer in the silica pore system. A multi-layer stationary-phase model is presented in which the first layer consists of an adsorbed monolayer of PMOS and the second layer is immobilized by gamma radiation.

A method for determination of bentazon residues in water has been developed. The method involves solid-phase extraction with C18 extraction tubes and high-performance liquid chromatographic analysis. A C18 column and guard column were used with UV detection at 230 nm, a mobile phase of methanol-water (60:40, v/v) at pH 4.6 (phosphoric acid) and a flow-rate of 0.8 ml/min. After optimization of the extraction and separation conditions, the method was validated. The method developed can be used for determination of bentazon in water, within the international limits of 0.1 microgram/l, with a 500-fold pre-concentration.