A continuous flow system combined with a sensing fluorimetric transductor for the determination of β-naphthol

A single automatic method for continuous flow determination of β-naphthol based on the enhancement of its native fluorescence once the analyte was transitorily retained on-line on a solid support (QAE A-25 resin) is reported. So, a flow-through optosensor was developed using a flow-injection analysis system with solid phase fluorimetric transduction. KCl (0.15 mol l⁻¹) at pH 12.0 was used as carrier solution. To obtain the optimum fluorescence signal the wavelengths chosen were 245 nm (excitation) and 420 nm (emission). The response of the sensor was directly proportional to the sample volume injected in the studied range 40-1500 μl. Approximately one higher order of magnitude is achieved in sensitivity when 1500 μl are used with respect to the use of 40 μl of sample. The sensor was calibrated for three different injection volumes: 40, 600 and 1500 μl, responding linearly in the measuring range of 2-60, 0.5-15 and 0.2-5 μg l⁻¹ with detection limits of 0.5, 0.09 and 0.05 μg l⁻¹, respectively. The relative standard deviation for ten independent determination is 0.6% (40 μl), 0.9% (600 μl) and 2.3% (1500 μl). A recovery study was performed onto three different spiked water samples at concentration levels from 1 to 2.5 μg l⁻¹ and the recovery percentage from the experimental data ranged between 101±2 and 105±5.     

Sensing of trace amounts of cadmium in drinking water using a single fluorescence-based optosensor

In this work, a single solid surface fluorescence based flow-through optosensor has been developed for the determination of trace amounts of cadmium in drinking water samples. The developed methodology is based on the transient immobilization of the target species on an appropriate active solid sensing zone (Sephadex QAE-A25).The target species was the fluorogenic chelate, formed as a result of the on-line complexation of Cd (II) with 8-hydroxyquinoline-5-sulfonic acid, being its fluorescence signal continuously monitored at an emission wavelength of 520 nm upon excitation at 360 nm. The effect of instrumental, chemical and flow-injection variables on the fluorescence signal were carefully investigated. Under optimized conditions, the proposed optosensor was calibrated in the range 2-60 μg l^− 1, obtaining a detection limit of 0.48 μg l^− 1, and a R.S.D of 1.9%, with a sampling frequency of 18 h^− 1. The proposed method was satisfactorily applied to different drinking water samples, with recoveries between 97% and 104%. Simplicity, low cost and easy operation are the main and more remarkable features of the proposed procedure.     

Fluorimetric detector and sensor for flow analysis made of light emitting diodes

Two compact optoelectronic fluorimetric devices operating according to the paired-emitter-detector-diode concept have been developed. The fluorimetric detector, fabricated of three light emitting diodes only, has been applied for the development of fluorimetric optosensor by further integration with sensing solid phase. In these investigations as a model analyte and as a model sensing layer useful for solid phase spectrometry, riboflavin and C18-silica have been chosen, respectively. Both developed analytical devices have been applied for non-stationary fluorimetric measurements performed under conditions of flow injection analysis. The presented flow-through detector and sensor operating under given flow conditions offer riboflavin determination in mg L⁻¹ and μg L⁻¹ ranges of concentration, respectively.     

Sensitive determination of hydrazine in water by gas chromatography–mass spectrometry after derivatization with ortho-phthalaldehyde

A gas chromatography–mass spectrometric (GC–MS) method has been established for the determination of hydrazine in drinking water and surface water. This method is based on the derivatization of hydrazine with ortho-phthalaldehyde (OPA) in water. The following optimum reaction conditions were established: reagent dosage, 40 mg mL⁻¹ of OPA; pH 2; reaction for 20 min at 70 °C. The organic derivative was extracted with methylene chloride and then measured by GC–MS. Under the established condition, the detection and the quantification limits were 0.002 μg L⁻¹ and 0.007 μg L⁻¹ by using 5.0-mL of surface water or drinking water, respectively. The calibration curve showed good linearity with r² = 0.9991 (for working range of 0.05–100 μg L⁻¹) and the accuracy was in a range of 95–106%, and the precision of the assay was less than 13% in water. Hydrazine was detected in a concentration range of 0.05–0.14 μg L⁻¹ in 2 samples of 10 raw drinking water samples and in a concentration range of 0.09–0.55 μg L⁻¹ in 4 samples of 10 treated drinking water samples.     

Multiwavelength fluorescence based optosensor for simultaneous determination of fuberidazole, carbaryl and benomyl

In the present work, a novel approach is proposed for the simultaneous determination of three widely used pesticides (namely, fuberidazole (FBZ), carbaryl (CBL) and benomyl (BNM)). The proposed method is based on a single continuous-flow solid surface fluorimetric multi-optosensor implemented with the use of a minicolumn placed just before the flow-through cell and filled with C₁₈ silica gel. The three pesticides are determined from an only injection (simultaneous determination): the minicolumn strongly retains two of them while the third develops a transitory signal when passing through the sensing solid microzone. Then, two alternate eluting solutions appropriately selected perform the sequential elution of the two pesticides from the minicolumn, achieving the detection zone and developing their transitory signals. The proposed optosensor works under optimal sensitivity conditions for all the three analytes because of the use of multi-wavelength fluorescence detection mode, so recording three different signals corresponding at three pairs of optima excitation/emission wavelengths. Using a sample volume of 2100 μl, the system was calibrated in the range 0.5-15, 40-800 and 50-1000 μg l⁻¹ with detection limits of 0.09, 6 and 9 μg l⁻¹ for FBZ, CBL and BNM, respectively. The R.S.D values (n=10) were lower than 2% in all cases. The proposed methodology was applied satisfactorily to water samples. Recovery percentages ranging from 97.8 to 101.1%, 97.9 to 103% and from 97 to 105% for FBZ, CBL and BNM, respectively, were obtained.     

Determination of dissolved sulphides in waste water samples by flow-through stripping chronopotentiometry with a macroporous mercury-film electrode

Sulphides in water samples were determined by stripping chronopotentiometry in a computer controlled flow system with a flow-through electrochemical cell. The working electrode was a porous glassy carbon electrode coated with Nafion and mercury. The sample was diluted with 0.1 mol L⁻¹ NaOH and analysed. Sulphides in the sample were collected in the porous electrode as mercury sulphide and then stripped by a current of −500 μA. The limit of detection was found to be 1.6 μg L⁻¹ and 0.5 μg L⁻¹ for 1 mL and 5 mL of preconcentrated sample, respectively. The linear range for 1 mL sample was found to be 5-400 μg L⁻¹. The repeatability and reproducibility was found to be 2.6% and 4.8%, respectively. The method was applied to analyses of waste water samples from a tannery.     

Analysis of phenylurea and propanil herbicides by solid-phase microextraction and liquid chromatography combined with post-column photochemically induced fluorimetry derivatization and fluorescence detection

This study examines the application of solid-phase microextraction coupled with high performance liquid chromatography combined with post-column photochemically induced fluorimetry derivatization and fluorescence detection (SPME-HPLC-PIF-FD) for the determination of four phenylurea herbicides (monolinuron, diuron, linuron and neburon) and propanil in groundwater. Direct immersion (DI) SPME was applied using a 60 μm polydimethylsiloxane/divinylbenzene (PDMS/DVB) fiber for the extraction of the pesticides from groundwater samples. An AQUASIL C₁₈ column (150 mm × 4.6 mm i.d., 5 μm) was used for separation and determination in HPLC. The method was evaluated with respect to the limits of detection (LODs) and the limits of quantification (LOQs) according to IUPAC. The limits of detection varied between 0.019 μg L⁻¹ and 0.034 μg L⁻¹. Limits of quantification ranged between 0.051 μg L⁻¹ and 0.088 μg L⁻¹. These values meet the recommended limits for individual pesticides in groundwater (0.1 μg L⁻¹) established by the EU. Recoveries ranged between 86% and 105% and relative standard deviation values between 2% and 8%.     

Exploiting the bead injection LOV approach to carry out spectrophotometric assays in wine: application to the determination of iron

A sequential injection lab-on-valve (SI-LOV) system was used to develop a new methodology for the determination of iron in wine samples exploiting the bead injection (BI) concept for solid phase extraction and spectrophotometric measurement. Nitrilotriacetic Acid (NTA) Superflow resin was used to build the bead column of the flow through sensor. The iron (III) ions were retained by the bead column and react with SCN⁻ producing an intense red colour. The change in absorbance was monitored spectrophotometrically on the optosensor at 480 nm. It was possible to achieve a linear range of 0.09-5.0 mg L⁻¹ of iron, with low sample and reagent consumption; 500 μL of sample, 15 μmol of SCN⁻, and 9 μmol of H₂O₂, per assay. The proposed method was successfully applied to the determination of iron in wine, with no previous treatment other than dilution, and to other food samples.     

A multicommuted fluorescence-based sensing system for simultaneous determination of Vitamins B2 and B6

A multicommuted flow-through optosensor based on the direct fluorescence measurements of Vitamins B₂ and B₆ using a non-polar sorbent (C₁₈ silica gel) as solid sensing zone (to accomplish the separation and subsequent preconcentration/detection of the target analytes) have been developed. The proposed flow system was controlled by Java-written home-made software and designed using three-way solenoid valves for independent automated manipulation of sample and carrier solutions. The native fluorescence signal was simultaneously monitored at two pairs of excitation/emission wavelengths (450/519 and 294/395 for B₂ and B₆, respectively). The separation of the analytes was performed in the detection flow cell, using the differences in the sorption/elution process on the solid support between the two vitamins, due to their different polarity. Using an optimised sampling time, the analytical signal showed linearity in the range 0.01-0.4 and 0.15-3 μg ml⁻¹ with detection limits of 0.003 and 0.045 μg ml⁻¹ for B₂ and B₆, respectively, obtaining R.S.D. (%) values better than 2% for both analytes. The proposed methodology was applied to different pharmaceutical preparations, obtaining remarkably good results with recoveries ranging from 96 to 107.5%.     

Validation of an enzyme-linked immunosorbent assay (ELISA) for the determination of 4-nonylphenol and octylphenol in surface water samples by LC-ESI-MS

4-Nonylphenol (NP) and octylphenol (OP) were measured by direct ELISA in both laboratory-fortified and surface water samples collected monthly from 10 rivers. In this procedure, samples were concentrated by solid phase extraction (SPE) using Lichrolut RP-18 sorbent with good recoveries obtained for both LC-MS and ELISA, giving a low level of detection (LOD) at the range of low μg L⁻¹ and good reproducibility. Analysis of 40 surface water samples demonstrated that the ELISA may be a useful screening tool for the determination of the alkylphenols in surface water matrices. The concentration of NP and OP in surface waters ranged from 0.11 to 6.58 μg L⁻¹. A good correlation of results obtained by ELISA and LC-MS within the concentration range of 0.08-6.86 μg L⁻¹ was found in the river samples [R² = 0.924, n = 39]. The influence of various factors on assay determination was also discussed.     

Speciation analysis of mercury in sediments, zoobenthos and river water samples by high-performance liquid chromatography hyphenated to atomic fluorescence spectrometry following preconcentration by solid phase extraction

A high-pressure microwave digestion was applied for microwave-assisted extraction (MAE) of mercury species from sediments and zoobenthos samples. A mixture containing 3 mol L⁻¹ HCl, 50% aqueous methanol and 0.2 mol L⁻¹ citric acid (for masking co-extracted Fe³⁺) was selected as the most suitable extraction agent. The efficiency of proposed extraction method was better than 95% with R.S.D. below 6%. A preconcentration method utilizing a “homemade” C18 solid phase extraction (SPE) microcolumns was developed to enhance sensitivity of the mercury species determination using on-column complex formation of mercury-2-mercaptophenol complexes. Methanol was chosen for counter-current elution of the retained mercury complexes achieving a preconcentration factor as much as 1000. The preconcentration method was applied for the speciation analysis of mercury in river water samples. The high-performance liquid chromatography-cold vapour atomic fluorescence spectrometric (HPLC/CV-AFS) method was used for the speciation analysis of mercury. The complete separation of four mercury species was achieved by an isocratic elution of aqueous methanol (65%/35%) on a Zorbax SB-C18 column (4.6 mm × 150 mm, 5 μm) using the same complexation reagent (2-mercaptophenol). The limits of detection were 4.3 μg L⁻¹ for methylmercury (MeHg⁺), 1.4 μg L⁻¹ for ethylmercury (EtHg⁺), 0.8 μg L⁻¹ for inorganic mercury (Hg²⁺), 0.8 μg L⁻¹ for phenylmercury (PhHg⁺).     

Ultra-sensitive determination of cyanide in surface water by gas chromatography–tandem mass spectrometry after derivatization with 2-(dimethylamino)ethanethiol

A gas chromatography–tandem mass spectrometric (GC–MS/MS) method has been established for the determination of cyanide in surface water. This method is based on the derivatization of cyanide with 2-(dimethylamino)ethanethiol in surface water. The following optimum reaction conditions were established: reagent dosage, 0.7 g L⁻¹ of 2-(dimethylamino)ethanethiol; pH 6; reaction carried out for 20 min at 60 °C. The organic derivative was extracted with 3 mL of ethyl acetate, and then measured by using GC–MS/MS. Under the established conditions, the detection and quantification limits were 0.02 μg L⁻¹ and 0.07 μg L⁻¹ in 10-mL of surface water, respectively. The calibration curve had a linear relationship relationship with y = 0.7140x + 0.1997 and r² = 0.9963 (for a working range of 0.07–10 μg L⁻¹) and the accuracy was in a range of 98–102%; the precision of the assay was less than 7% in surface water. The common ions Cl⁻, F⁻, Br⁻, NO₃⁻, SO₄²⁻, PO₄³⁻, K⁺, Na⁺, NH₄⁺, Ca²⁺, Mg²⁺, Ba²⁺, Mn⁴⁺, Mn²⁺, Fe³⁺, Fe²⁺ and sea water did not interfere in cyanide detection, even when present in 1000-fold excess over the species. Cyanide was detected in a concentration range of 0.07–0.11 μg L⁻¹ in 6 of 10 surface water samples.     

Quantitative detection of tetracycline residues in honey by a simple sensitive immunoassay

Tetracyclines (TCs) are widely used for prevention and control of infectious diseases and have a great activity against variety of Gram-positive and Gram-negative bacteria. Due to the widespread use of TCs in animal husbandry, it can lead to an increase the risk of TCs remaining in human food. To protect consumers, many countries have set acceptable tolerance levels for these drugs. Therefore, it is necessary to establish a suitable analytical technique with specificity, sensitivity and simplicity.The biotin-avidin mediated ELISA method was performed to determine TC residues in honey quantitatively. By using PBS-EDTA assay buffer at pH 7.2, a honey solution of TC standard was prepared and diluted. And no additional pre-treatment of sample was required in this method. The limit of detection and limit of quantitation of the optimized method were 3.98 × 10⁻¹⁰ M (0.19 μg L⁻¹) and 7.94 × 10⁻¹⁰ M (0.38 μg L⁻¹), respectively, and the dynamic range was from 1.52 μg L⁻¹ to 152 μg L⁻¹ of TC in honey. No cross-reactivity was observed with the structurally similar compounds, and mean percent recoveries of TC spiked in honey ranged from 95% to 101%. Compared to other methods, this method was superior in terms of detection limit, dynamic range, and % recovery with simple sample-preparation.     

Speciation of inorganic arsenic in a sequential injection dual mini-column system coupled with hydride generation atomic fluorescence spectrometry

The separation and speciation of inorganic arsenic(III) and arsenic(V) are facilitated by employing a novel sequential injection system incorporating two mini-columns followed by detection with hydride generation atomic fluorescence spectrometry. An octadecyl immobilized silica mini-column is used for selective retention of the complex between As(III) and APDC, while the sorption of As(V) is readily accomplished by a 717 anion exchange resin mini-column. The retained As(III)-PDC complex and As(V) are effectively eluted with a 3.0 mol L⁻¹ hydrochloric acid solution as stripping reagent, which well facilitates the ensuing hydride generation process via reaction with tetrahydroborate. With a sampling volume of 1.0 mL and an eluent volume of 100 μL for both species, linear ranges of 0.05-1.5 μg L⁻¹ for As(III) and 0.1-1.5 μg L⁻¹ for As(V) are obtained, along with enrichment factors of 7.0 and 8.2, respectively. Precisions of 2.8% for As(III) and 2.9% for As(V) are derived at the concentration level of 1.0 μg L⁻¹. The practical applicability of the procedure has been demonstrated by analyzing a certified reference material of riverine water (SLRS-4), in addition to spiking recovery in a lake water sample matrix.     

Determination of environmental organic pollutants with a portable optical immunosensor

A portable surface plasmon resonance (SPR) optical biosensor device is described as a direct immunosensing system to determine organic pollutants in natural water samples. Monitoring of organochlorine (DDT), organophosphorus (chlorpyrifos) and carbamate (carbaryl) compounds within the concentration levels stipulated by the European legislation, can be accomplished using this immunosensor. The lowest limit of detection (LOD) was obtained for DDT, at 20 ng L⁻¹, whilst 50 ng L⁻¹ and 0.9 μg L⁻¹, were achieved for chlorpyrifos and carbaryl, respectively. Matrix effects were evaluated for the carbaryl immunoassay in different water types with detection limits within the range of carbaryl standard curves in distilled water (0.9-1.4 μg L⁻¹). The covalent immobilization of the analyte derivative through an alkanethiol self-assembled monolayer (SAM) allowed the reusability of the sensor surface during more than 250 regeneration cycles. The quality of the regeneration was proved over a 1-month period of continuous working. The analysis time for a complete assay cycle, including regeneration, comprises 24 min. Our portable SPR-sensor system is already a market product, commercialized by the company SENSIA, SL. The size and electronic configuration of the device allow its portability and utilization on real contaminated locations.     

Determination of the steroid hormone levels in water samples by dispersive liquid-liquid microextraction with solidification of a floating organic drop followed by high-performance liquid chromatography

In this study, the steroid hormone levels in river and tap water samples were determined by using a novel dispersive liquid-liquid microextraction method based on the solidification of a floating organic drop (DLLME-SFO). Several parameters were optimized, including the type and volume of the extraction and dispersive solvents, extraction time, and salt effect. DLLME-SFO is a fast, cheap, and easy-to-use method for detecting trace levels of samples. Most importantly, this method uses less-toxic solvent. The correlation coefficient of the calibration curve was higher than 0.9991. The linear range was from 5 to 1000 μg L⁻¹. The spiked environmental water samples were analyzed using DLLME-SFO. The relative recoveries ranged from 87% to 116% for river water (which was spiked with 4 μg L⁻¹ for E1, 3 μg L⁻¹ for E2, 4 μg L⁻¹ for EE2 and 9 μg L⁻¹ for E3) and 89% to 102% for tap water (which was spiked with 6 μg L⁻¹ for E1, 5 μg L⁻¹ for E2, 6 μg L⁻¹ for EE2 and 10 μg L⁻¹ for E3). The detection limits of the method ranged from 0.8 to 2.7 μg L⁻¹ for spiked river water and 1.4 to 3.1 μg L⁻¹ for spiked tap water. The methods precision ranged from 8% to 14% for spiked river water and 7% to 14% for spiked tap water.     

Multiplex competitive microbead-based flow cytometric immunoassay using quantum dot fluorescent labels

In answer to the ever-increasing need to perform the simultaneous analysis of environmental hazards, microcarrier-based multiplex technologies show great promise. Further integration with biofunctionalized quantum dots (QDs) creates new opportunities to extend the capabilities of multicolor flow cytometry with their unique fluorescence properties. Here, we have developed a competitive microbead-based flow cytometric immunoassay using QDs fluorescent labels for simultaneous detection of two analytes, bringing the benefits of sensitive, rapid and easy-of-manipulation analytical tool for environmental contaminants. As model target compounds, the cyanobacterial toxin microcystin-LR and the polycyclic aromatic hydrocarbon compound benzo[a]pyrene were selected. The assay was carried out in two steps: the competitive immunological reaction of multiple targets using their exclusive sensing elements of QD/antibody detection probes and antigen-coated microsphere, and the subsequent flow cytometric analysis. The fluorescence of the QD-encoded microsphere was thus found to be inversely proportional to target analyte concentration. Under optimized conditions, the proposed assay performed well within 30 min for the identification and quantitative analysis of the two environmental contaminants. For microcystin-LR and benzo[a]pyrene, dose–response curves with IC₅₀ values of 5 μg L⁻¹ and 1.1 μg L⁻¹ and dynamic ranges of 0.52–30 μg L⁻¹ and 0.13–10 μg L⁻¹ were obtained, respectively. Recovery was 92.6–106.5% for 5 types of water samples like bottled water, tap water, surface water and seawater using only filtration as sample pretreatment.     

Implementation of multicommutation principle with flow-through multioptosensors

For the first time, the combination of multicommutation concept with flow-through multioptosensors is exploited and a biparameter multicommutated UV sensor is developed. A very easy-operated and automated sensing device is proposed here and demonstrated to be useful in the routine analysis by applying it to the determination of two widely used pharmaceuticals chosen as model analytes: salicylamide and caffeine. The particulated solid phase used performs two functions: (a) states selectivity conditions by itself in the flow cell and (b) provides appropriate separation of the analytes in the on-line precolumn, thus making possible the sequential arrival of the analytes to the detection zone. Both carrier and eluting solutions chosen allow (with a very simple solenoid valve configuration and operation) the transitory signals to be developed in a completely automated device. Drastic reduction of carrier and eluting solutions used are achieved as compared to typically FIA optosensor configuration. The sensor responds linearly in the range 2-30 and 1-14 μg ml⁻¹ for SLC and CF, with detection limits of 0.33 and 0.15 μg ml⁻¹, respectively. Solution savings between 50 and 85% were obtained when comparing to typical flow injection analysis (FIA).     

Sensitive and ultra-fast determination of arsenic(III) by gas-diffusion flow injection analysis with chemiluminescence detection

A novel chemiluminescence gas-diffusion flow injection system for the determination of arsenic(III) in aqueous samples is described. The analytical procedure involves injection of arsenic(III) samples and standards into a 0.3 mol L⁻¹ hydrochloric acid carrier stream which is merged with a reagent stream containing 0.2% (w/v) sodium borohydride and 0.015 mol L⁻¹ sodium hydroxide. Arsine, generated in the combined carrier/reagent donor stream, diffuses across the hydrophobic Teflon membrane of the gas-diffusion cell into an argon acceptor stream and then reacts with ozone in the flow-through chemiluminescence measuring cell of the flow system. Under optimal conditions, the method is characterized by a wide linear calibration range from 0.6 μg L⁻¹ to 25 mg L⁻¹, a detection limit of 0.6 μg L⁻¹ and a sample throughput of 300 samples per hour at 25 mg L⁻¹ and 450 samples per hour at 25 μg L⁻¹.     

Biosensor immunoassays for the detection of bisphenol A

Bisphenol A (BPA) is a xenoestrogen found in the environment, in consequence, for the biosensor detection of BPA we raised antibodies (polyclonal (PAbs) and monoclonal (MAbs)) against a structural analogue of BPA, 4,4 bis-(4-hydroxyphenyl) valeric acid (BVA). The kinetics of the MAb-BPA interaction were evaluated and the MAb providing the highest affinity was directly immobilized onto the sensor chip surface to evaluate a direct assay. Afterwards, the performance of the MAbs and the PAbs was compared in an inhibition assay using a BVA-coated chip.The highest sensitivity (limit of detection (LOD) of 0.4 μg L⁻¹) was obtained with MAb 12 in the direct assay. However, the inhibition assay was the most robust and the PAbs showed the highest sensitivity (LOD of 0.5-1 μg L⁻¹). The antibodies were specific for BVA and BPA as only minor cross-reactivities were found toward structurally related compounds or other endocrine disruptors. In the inhibition assay (with a run time of 6 min), water samples spiked with BPA at different levels (0.5-50 μg L⁻¹) resulted in recoveries varying between 68% and 121%. The sensitivity of the inhibition assay could be improved 40 times (LOD of 0.03 μg L⁻¹ with the Mab 12-based assay) using solid phase extraction (SPE).