Determination of salbutamol using on-line solid-phase extraction and sequential injection analysis. Comparison of chemiluminescence and fluorescence detection

Determination of salbutamol using sequential injection analysis (SIA) with chemiluminescence and fluorescence detection has been devised. The chemiluminescence signal was emitted during the oxidation of salbutamol by potassium permanganate in sulfuric acid medium. Sodium polyphosphate was used as chemiluminescence enhancer. The fluorescence signal (excitation wavelength 230 nm) was also measured in sulfuric acid medium. Both detection techniques were compared with respect to the application of the methods to the determination of salbutamol in biological materials. The sample pre-treatment takes place directly in the SIA system, when salbutamol is adsorbed on the solid-phase (Baker-carboxylic acid) microcolumn integrated into the system. Sulfuric acid serves both as the reagent and the eluent. The lab-made SIA system consisted of a 2.5-mL Cavro syringe pump, ten-port Vici Valco selection valve and Spectra-Physics FS 970 fluorescence detector, which was lab-modified for chemiluminescence detection. The system was controlled by a PC using originally compiled LabVIEW-supported software. Concentrations, volumes of reagents and flow rates were optimised by a simplex method. Salbutamol was determined in the linear range 0.05-10 microg mL(-1) (RSD 1.53%), with the detection limit (3 sigma) 0.03 microg mL(-1) and sample throughput of 42 samples per hour with chemiluminescence detection in standard solutions. The fluorescence detection enabled the determination of salbutamol in standard solutions in the linear range 0.5-100 microg mL(-1) (RSD 2.69%), with the detection limit 0.2 microg mL(-1) and sample throughput of 24 h(-1). The proposed methods were applied to the determination of salbutamol in human serum and urine. However, serum is a very complicated matrix and the SIA-SPE analysis did not provide satisfactory results. It was possible to determine salbutamol in human urine using this technique. Better recovery was achieved with fluorescence detection.     

Comparison of traditional cloud-point extraction and on-line flow-injection cloud-point extraction with a chemiluminescence method using benzo[a]pyrene as a marker

In this work, using benzo(a)pyrene (BaP) as marker, the analytical merits of on-line flow-injection cloud-point extraction (FI CPE), including preconcentration factor, extraction efficiency, sample throughput, and analysis time were evaluated by use of peroxyoxalate chemiluminescence (CL) detection. Moreover, by detailed discussion of several preconcentration conditions for traditional and on-line FI CPE the advantages of on-line FI CPE became conspicuously apparent. When coupled with separation techniques such as high-performance liquid chromatography (HPLC) or capillary electrophoresis (CE), on-line FI CPE–CL has much potential for analysis of low concentrations of polycyclic aromatic hydrocarbons (PAH) in environmental samples.     

Micelle-sensitized constant-energy synchronous fluorescence spectrometry for the simultaneous determination of pyrene, benzo[a]pyrene and perylene.

The use of micellar media in constant-energy synchronous fluorescence spectrometry has been proposed. The influence of some aqueous micellar systems on the determination of pyrene, perylene and benzo[a]pyrene has been investigated. The presence of these micellar systems allows their determination in aqueous media, thus avoiding the use of an organic solvent, and greatly enhances the fluorescence signals. The combination of a constant-energy synchronous scanning technique and a micellar system provided a single spectrum for the simultaneous identification and quantitative determination of the three polycyclic aromatic hydrocarbons (PAHs). Further there was no energy transfer among them, making the measurement simple and fast. A constant-energy difference of 2800 cm(-1) was selected. The analytical characteristics of the proposed method in the presence of anionic micelles of sodium dodecylsulfate (SDS) were studied. The detection limits were at a level of ng ml(-1). Analysis of water samples from two different origins spiked with known amount of pyrene, perylene and benzo[a]pyrene also gave satisfactory results, and total average recoveries were greater than 97.1%.     

Determination of benzo[a]pyrene in shale oil-by solid-surface fluorescence

A method has been developed for determining benzo[a]pyrene in shale oil by combining dry-column chromatography, thin-layer chromatography and fluorescence spectrometry. A two-step separation method was employed to isolate benzo[a]pyrene from shale oil. Benzo[a]pyrene was identified and determined by detecting its fluorescence directly from chromatoplates; as little as 0.06 ng can be detected. The limit of detection of benzo[a]pyrene in shale oil is ca. 1.2 ppm and the reproducibility of the method is ±2.6 ppm.     

A simple method for the determination of benzo[a]pyrene and benzo[k]fluoranthene in a mixture

Atmospheric particulate matter is routinely analysed for the concentration of the carcinogen benzo[a]pyrene (BaP). Benzo[k]fluoranthene (BkF) has identical spectral characteristics and interferes in the fluorimetric estimation. A simple method for the determination of BaP in the presence of BkF is described. Based on quenching of the fluorescence of BaP by the addition of small amounts of nitromethane, the method can be used for accurate estimation for wide concentration ratios of BaP to BkF ranging from 0.1 to 10.0.     

Supersonic jet fluorescence spectrometry of perylene and benzo[a]pyrene

A supersonic jet instrument for fluorescence spectrometry is described. It consists of a high-temperature free expansion nozzle for continuous sample introduction and a vacuum chamber equipped with a high-speed pumping system. Rotationally cooled spectra obtained with the supersonic jet are compared with gas-phase spectra measured at high temperature for perylene and benzo[a]pyrene molecules. Each component of the unresolved band structure in the high-temperature spectra was found to be composed of a rotational congestion of several vibrational bands. For a 1:1 mixture of perylene and benzo[a]pyrene, selective detection is possible by using supersonic jet spectrometry. The detection limit for perylene is 100 ng. The advantage of this technique over other low-temperature spectrometric methods based on Shpol'skii and matrix isolation effects are discussed.     

Humic acid-bonded silica as a novel sorbent for solid-phase extraction of benzo[a]pyrene in edible oils

A novel solid-phase extraction (SPE) sorbent, humic acid-bonded silica (HAS), was prepared. Humic acids (HAs) were grafted onto silica matrices via an amide linkage between humyl chloride and the amido terminus of 3-aminopropyltrimethoxysilane (APTS)-silica gel. The resulting material was characterized by Fourier transform infrared spectrometer, elemental analysis, and nitrogen adsorption analysis. This sorbent exhibits an excellent adsorption capacity for some electron-abundant analytes owing to its peculiar structure. In this paper, we choose benzo[a]pyrene (BaP) in oil as a probe to validate the adsorption capacity of the material. Thus a fast, cheap and simple SPE method with humic acid-bonded silica cartridge for edible oil clean-up, followed by high-performance liquid chromatography (HPLC) with fluorescence detection was established. The effects of experimental variables, such as washing and elution solvents, and the amount of sorbents have been studied. The recoveries of BaP in edible oils spiked at 0.2-100 μg kg⁻¹ were in the range of 78.8-102.7% with relative standard deviations ranging between 1.3 and 9.3%; the limit of detection was -0.06 μg kg⁻¹.     

Detection of DNA adducts of benzo[a]pyrene using immunoelectrophoresis with laser-induced fluorescence. Analysis of A549 cells

Detection of benzo[a]pyrene diol epoxide (BPDE)-damaged DNA in a human lung carcinoma cell line (A549) has been performed using free zone affinity capillary electrophoresis with laser-induced fluorescence (LIF). Using BPDE as a model carcinogenic compound, the speed, sensitivity and specificity of this technique was demonstrated. Under free zone conditions, an antibody bound adduct was baseline-resolved from an unbound adduct in less than 2 min. The efficiencies of separation were in excess of 6 x 10(5) and 1 x 10(6) plates per meter for the antibody-bound and unbound adducts, respectively. Separation using a low ionic strength buffer permitted the use of a high electric field (830 V/cm) without the loss of resolving power. Using LIF detection, a concentration detection limit of roughly 3 x 10(-10) M was achieved for a 90-mer oligonuleotide containing a single BDPE. The use of formamide in the incubation buffer to enhance denaturing of DNA did not affect the stability of the complex between the antibody and the adducts. Using a fluorescently labeled BPDE-modified DNA adduct probe, a competitive assay was established to determine the levels of BPDE-DNA adducts in A549 cells.     

Effects of cluster formation on spectra of benzo[a]pyrene and benzo[e]pyrene

Absorption and fluorescence emission spectra of the polycyclic aromatic hydrocarbons benzo[a]pyrene (BaP) and benzo[e]pyrene (BeP) in solution and adsorbed on silica have been obtained and compared to examine the spectroscopic effects of clustering. Molecular mechanics calculations with the UFF potential were done to optimize monomer, dimer and trimer geometries, and energy differences were determined by MP2/6-31G* calculations. Fluorescence emission spectra of adsorbed BeP and BaP display a red shift that progresses with increased loading, and the two differ in their photodegradation kinetics. The experimental and theoretical results are found to be consistent.     

Second-derivative constant-wavelength synchronous scan spectrofluorimetry for determination of benzo[b]fluoranthene, benzo[a]pyrene and indeno[1,2,3-cd]pyrene in drinking water

The use of derivative constant-wavelength synchronous scan fluorimetry is reported for the determination of three polycyclic aromatic hydrocarbon pollutants in drinking water (linearity range 0.4-4 mug 1(-1)). The limits of detection (LD) and quantification (LQ) (mug 1(-1)) are 0.01 and 0.07 for benzo[b]fluoranthene, 0.03 and 0.12 for benzo[a]pyrene and 0.19 and 0.57 for indeno[1,2,3-cd]pyrene in the presence of three other pollutants, benzo[k]fluoranthene, benzo[ghi]perylene and fluoranthene. The precision (RSD /= 85%) were satisfactory.     

The selective cleanup of complex matrices and simultaneous separation of benzo[a]pyrene by solid-phase extraction with MgO microspheres as sorbents

A new method for the selective cleanup of complex matrices and simultaneous separation of benzo[a]pyrene (BaP) was developed in this study. This method was based on solid-phase extraction (SPE) using magnesium oxide microspheres as sorbents, and it eliminated interferences from various impurities, such as lipids, sulphur, pigments, halobenzenes, polychlorodibenzo-p-dioxins and polychlorodibenzofurans. Several parameters, including the volume of rinsing and eluting solvents, the type of loading solvents and SPE sorbents, were optimized systematically. The capability for impurity removal was verified by gel permeation chromatography, gas chromatography, and liquid chromatography. Compared to commercial sorbents (silica gel, florisil and alumina), MgO microspheres exhibited excellent performance in the selective isolation of BaP and removal of impurities. The proposed method was applied to detect BaP in complex samples (sediments, soils, fish, and porcine liver). The limit of quantification (LOQ) was 1.04 ngL(-1), and the resulting regression coefficient (r(2)) was greater than 0.999 over a broad concentration range (9.5-7600 ngL(-1)). In contrast to traditional methods, the proposed method can give rise to higher recovery (85.1-100.8%) and better selectivity with simpler operation and less consumption of organic solvents (20-40 mL).     

A convenient new synthesis of benzo[a]pyrene

A convenient new synthesis of the ubiquitous environmental carcinogen benzo[a]pyrene (BaP) is described. In the key step, the method entails Suzuki coupling of naphthalene 2-boronic acid with 2-bromobenzene-1,3-dialdehyde and requires only three steps. It is considerably shorter and simpler than the older methods and provides BaP in higher overall yield.     

Reinvestigation of the jutz synthesis of benzo [e] pyrene and benzo [a] pyrene derivatives from benzanthrene

Base-catalyzed reactions of benzanthrene with “vinamidinium salts” ($\text{2a-c}$ ) followed by thermal electrocyclic ring closure are regiospecific affording only benzo [e] pyrene derivatives, contrary to previous claims.     

Multi-syringe flow injection solid-phase extraction system for on-line simultaneous spectrophotometric determination of nitro-substituted phenol isomers

In this paper, a time-based multi-syringe flow injection (MSFI) approach is proposed for automated disk-based sorbent extraction of three nitro-substituted phenol isomers (2-, 3-, and 4-nitrophenol) followed by on-line simultaneous determination of individual species by diode-array spectrophotometry. The method involves the on-line enrichment of the targeted analytes from an acidic medium containing 0.1 mol L⁻¹ HCl onto a co-polymeric sorbent material, and the concurrent removal of potentially interfering matrix components. The nitrophenol isomers are subsequently eluted with an alkaline solution (0.7 mol L⁻¹ NaOH), whereupon the eluate is delivered to a diode-array spectrophotometer for recording of the spectral data in the UV-vis region. Deconvolution of strongly overlapped spectra was conducted with multivariate regression models based on multiple linear regression calibration. The analytical performance of the chemometric algorithm was characterized by relative prediction errors and recoveries.The MSFI manifold was coupled to a multiposition selection valve to set a rugged analyzer that ensures minimum operational maintenance via exploitation of membrane switching protocols. As compared with earlier methods for isolation/pre-concentration of nitro-substituted phenols based on liquid-liquid extraction, the proposed flow-through disk-based system should be regarded as an environmentally friendly approach because the use of harmful organic solvents is circumvented. Under the optimized chemical and physical variables, the 3σ_blank detection limits for 2-, 3-, and 4-nitrophenol were 1.2, 3.2 and 0.3 μmol L⁻¹ for a sample loading volume of 1.5 mL, and the relative standard deviations were ≤5.0%. The flowing system, which is able to handle up to 135 samples automatically, was proven suitable for monitoring trace levels of the target isomers in mineral, tap, and seawater.     

Simultaneous determination of fluorophores with overlapped spectra by sequential injection analysis coupled to variable angle scanning fluorescence spectrometry and multivariate linear regression algorithms

An automated and versatile sequential injection spectrofluorimetric procedure for the simultaneous determination of multicomponent mixtures in micellar medium without prior separation processes is reported. The methodology is based upon the segmentation of a sample slug between two different buffer zones in order to attain both an improvement of sensitivity and residual minimization for the whole species. Resolution of overlapping fluorescence profiles is achieved using a variable angle scanning technique coupled to multivariate least-squares regression (MLR) algorithms at both sample edges.The potentialities of the described methodology are illustrated with the spectrofluorimetric determination of four widespread pesticides with different acid-base properties; viz. carbaryl (CBL) (1-naphthyl-N-methylcarbamate), fuberidazole (FBZ) (2-(2′-furyl)benzimidazole), thiabendazole (TBZ) (2-(4′-thiazolyl)benzimidazole) and warfarin (W) (3-α-acetonylbenzyl)-4-hydroxycoumarin). Detection limits at the 3σ level were 3.9, 0.02, 0.03 and 10 μg l⁻¹ for CBL, FBZ, TBZ and W, respectively at the maximum sensitivity pH. Dynamic ranges of 13-720 μg l⁻¹ CBL, 0.10-14 μg l⁻¹ FBZ, 0.19-60 μg l⁻¹ TBZ and 0.05-5 mg l⁻¹ W were achieved. Relative standard deviations (n=10) were 0.2% for 100 μg l⁻¹ CBL and 2.4 μg l⁻¹ FBZ, 0.7% for 8 μg l⁻¹ TBZ and 1.0% for 1 mg l⁻¹ W. The proposed automated methodology, which handles 17 samples/h, was validated and applied to spiked real water samples with very satisfactory results.     

Enrichment of benzo[a]pyrene in vegetable oils and determination by HPLC-FL

We have developed a simple method for the determination of the carcinogen Benzo[a]pyrene (BP) in vegetable oils. The method consists of extraction of the vegetable oil in acetonitrile, concentration to dryness in rotary evaporator and redissolution of the residue in hexane. The purification of the hexane extract was on Sep-Pack Silica Plus cartridges, and the determination of the BP in the isolated extract was by HPLC-FL. Detection and quantification limits were 0.23 and 0.32 μg kg⁻¹ of olive oil, respectively. Recovery (>93%) and RSD (<4%) were satisfactory. When applied to 18 oil samples, BP levels varied from not detected to 1.99 μg kg⁻¹.     

Automated solid-phase extraction hyphenated to voltammetry for the determination of quercetin using magnetic nanoparticles and sequential injection lab-on-valve approach

In this study, an automated sequential injection lab-on-valve (SI-LOV) system was designed for the on-line matrix removal and preconcentration of quercetin. Octadecyl functionalized magnetic silica nanoparticles were prepared and packed into the microcolumn of the LOV as adsorbents. After being adsorbed through hydrophobic interaction, the analyte was eluted and subsequently introduced into the electrochemical flow cell by voltammetric quantification. The main parameters affecting the performance of solid-phase extraction, such as sample pH and flow rate, eluent solution and volume, accumulation potential and accumulation time were investigated in detail. Under the optimum experimental conditions, a linear calibration curve was obtained in the range of 1.0 × 10(-8) to 1 × 10(-5) mol L(-1) with R(2) = 0.9979. The limit of detection (LOD) and limit of quantitation (LOQ) were 1.3 × 10(-9) and 4.3 × 10(-9) mol L(-1), respectively. The relative standard deviation (RSD) for the determination of 1.0 × 10(-6) mol L(-1) quercetin was found to be 2.9% (n = 11) along with a sampling frequency of 40 h(-1). The applicability and reliability of the automated method described here had been applied to the determination of quercetin in human urine and red wine samples through recovery experiments, and the obtained results were in good agreement with those obtained by the HPLC method.     

A selective method for the determination of benzo[a]pyrene in soil using porous graphitic carbon liquid chromatography columns

A back-flushing procedure using porous graphitic carbon (PGC) HPLC columns has been used successfully for the cleanup of soil samples for the determination of benzo[a]pyrene in ppb levels by an ODS-fluorescence HPLC column. The procedure was tested on nine random soil samples taken from an industrial area of the Kingdom of Bahrain. The mean percent recovery from the PGC column was 96% and the average coefficient of variation for the whole method was 5.2%.