Célia G. Amorim

Exploiting sequential injection analysis with lab-on-valve and miniaturized potentiometric detection Epinephrine determination in pharmaceutical products.

Miniaturized potentiometric units were constructed, evaluated and incorporated in a SIA-LOV manifold in order to the control of pharmaceutical analysis. The method validation was done with ephinefrine determinations in commercial pharmaceutical products. The optimization procedures were directed at potential versus epinephrine concentration. This approach was achieved by selecting 60cm of reactor and a flow rate of 11muLs(-1) and injecting 78muL of epinephrine standard solutions in a 1.0x10(-3)molL(-1) IO(4)(-) solution. A linear range was found for epinephrine concentrations between 2.0x10(-4) and 2.5x10(-3)molL(-l) with a slope of 35528mVLmol(-l) and r(2)=0.997. Under these conditions the analytical results for the commercial pharmaceutical formulations of 0.908 and 0.454mgmL(-1), respectively, with a R.S.D. of 0.34 for both, was obtained. Through down scaling periodate-selective electrode it was possible to benefit from the recognized advantages of the lab-on-valve sequential-injection based systems, namely regarding equipment portability, reduced consumption of the sample and the reagents and the reduction of effluent waste. Furthermore, the new periodate electrode configuration is easy to achieve in common laboratories and enables the implementation of low volume detection cell where the electrical noise, that is frequently presented in potentiometric based procedures, is significantly reduced.

Cyclodextrin-based potentiometric sensors for midazolam and diazepam.

In this work the implementation of benzodiazepine ion-selective electrodes for pharmaceutical formulations control is described. The solid-contact electrodes for midazolam and diazepam are based on polymeric membranes incorporating respectively beta-cyclodextrin and (2-hydroxiproyl)-gamma-cyclodextrin as ionophores, 2-fluorophenyl 2-nitrophenyl ether as plasticizer and potassium tetrakis (p-chlorophenyl) borate as ionic additive. For conventionally shaped midazolam electrode a slope of 61.9+/-1.3 mVdec(-1), a LLLR of 5.7+/-2.7 x 10(-4)gL(-1) and pH range of 2.6-5.4 was obtained, while the corresponding values for diazepam electrodes were of 67.6+/-3.0 mVdec(-1), 4.9+/-1.5 x 10(-2)gL(-1) and 1.9-2.7 pH units, respectively. Membrane optimization was based on the molar ratio between the ionophore and additive for midazolam and on inclusion cavity of cyclodextrin for diazepam. The miniaturization of the above-described electrodes gave rise to potentiometric detectors for sequential-injection lab-on-valve system with similar characteristics albeit the useful lifetime shortened from 1 year to approximately 15 days under continuous operation. The optimized flow conditions were achieved for sample injection volumes of 20 microL propelled towards the detection cell at the flow rate of 16 microLs(-1) during 80s. Real sample analysis revealed statistical accuracy and between-days precision comparable to the general used chromatographic-based procedure.

A SO2-selective electrode based on a Zn-porphyrin for wine analysis.

This work describes the assessment of a SO2-selective electrode based on the use of the neutral carrier 5,10,15,20-tetraphenyl(porphyrinate)zinc(II) in a PVC membrane plasticized with 2-nitrophenyl phenyl ether. After being conditioned in 2 mol L(-1) diethylamine solution for 24 h, the electrode exhibited selective anionic response toward the analyte in a concentration interval of more than four decades, with an slope of -59.5 mV dec(-1), a practical detection limit of 3.7×10(-6) mol L(-1) and a low limit of linear range of 7.2×10(-6) mol L(-1). The response mechanism is based on the displacement of the diethylamine:metalloporphyrin complex equilibrium within membrane bulk, inducing a variation in the cationic-sites to ionophore ratio. In turn, free hydroxyl ions are complexed by the displaced ionophore in a ratio 1:1 and translated as single negative charge nernstian response. Finally, the selectivity of the electrode is evaluated in view of its application to wine analysis. Results had high accuracy and precision when compared with a reference method.

Fluorescence probe for mercury(II) based on the aqueous synthesis of CdTe quantum dots stabilized with 2-mercaptoethanesulfonate

Manipulation of the QD surface by changing morphology and surface capping ligands, as well as adjusting nanocrystal size, plays a crucial role on the selectivity and sensitivity exhibited by QDs towards a given target analyte. In this study, a novel aqueous synthesis of CdTe QDs capped with a thiol compound containing a sulfonate (SO3−) terminal group, viz., 2-mercaptoethanesulfonate (MES), was thoroughly investigated with the aim of obtaining a fluorescent probe for chemical analysis. The results obtained with the prepared CdTe-MES QDs in the determination of various metal ions demonstrated the high efficiency of these nanomaterials for determining Hg(II) levels. Upon optimization, linear working calibration curves for Hg(II) concentrations of up to 0.5 μmol L−1 were obtained, with a determination coefficient of 0.9984. The detection limit was 0.0095 μmol L−1 and the quantification limit was 0.0324 μmol L−1. When applied to the determination of Hg(II) in tap water samples, the developed method provided analytical results similar to those obtained with a reference method. The accuracy and precision of both methods were comparatively evaluated using the Students t-test and the Fischer test, and the tabulated values showed a good agreement at a 95% confidence level.

Fully automated analytical procedure for propofol determination by sequential injection technique with spectrophotometric and fluorimetric detections

In this work, an application of an enzymatic reaction for the determination of the highly hydrophobic drug propofol in emulsion dosage form is presented. Emulsions represent a complex and therefore challenging matrix for analysis. Ethanol was used for breakage of a lipid emulsion, which enabled optical detection. A fully automated method based on Sequential Injection Analysis was developed, allowing propofol determination without the requirement of tedious sample pre-treatment. The method was based on spectrophotometric detection after the enzymatic oxidation catalysed by horseradish peroxidase and subsequent coupling with 4-aminoantipyrine leading to a coloured product with an absorbance maximum at 485 nm. This procedure was compared with a simple fluorimetric method, which was based on the direct selective fluorescence emission of propofol in ethanol at 347 nm. Both methods provide comparable validation parameters with linear working ranges of 0.005-0.100 mg mL(-1) and 0.004-0.243 mg mL(-1) for the spectrophotometric and fluorimetric methods, respectively. The detection and quantitation limits achieved with the spectrophotometric method were 0.0016 and 0.0053 mg mL(-1), respectively. The fluorimetric method provided the detection limit of 0.0013 mg mL(-1) and limit of quantitation of 0.0043 mg mL(-1). The RSD did not exceed 5% and 2% (n=10), correspondingly. A sample throughput of approx. 14 h(-1) for the spectrophotometric and 68 h(-1) for the fluorimetric detection was achieved. Both methods proved to be suitable for the determination of propofol in pharmaceutical formulation with average recovery values of 98.1 and 98.5%.

Vortex-assisted liquid-liquid micro-extraction and high-performance liquid chromatography for a higher sensitivity methyl methacrylate determination in biological matrices

A vortex-assisted liquid-liquid micro-extraction coupled with high-performance liquid chromatography, with UV-vis, is proposed to pre-concentrate methyl methacrylate and to improve separation in biological matrices. The use of 1-octanol as extracting phase, its volume, the need for a dispersant agent, the agitation conditions and the cooling time before phase separation were evaluated. In optimum conditions, enrichment factors of 20 (±0.5) and enrichment recovery of 99% were obtained. The straightforward association of this extraction process with the HPLC method, previously regulated by the International Organization for Standardization, afforded a detection limit of 122 ng/mL and a quantification limit of 370 ng/mL. The within-batch precision, relative standard deviation, was 3% for a sample with 1.49 µg/mL and 4% for a sample with 13.4 µg/mL. The results showed a between batch-precision of 21% for experiments performed on five different days, for a sample with a concentration of 1.10 µg/mL in methyl methacrylate.

Biosensing based on pencil graphite electrodes

Pencil leads have been increasingly used as electrode material in electrochemical applications. Commonly denominated as pencil graphite electrodes (PGE), they represent a viable alternative to other standard electrodes due to their comparable electrical properties but mainly for their low cost and availability, enabling disposable applications. In order to achieve the best analytical performance literature evidences the type of lead (hardness level) and electrode surface pre-treatment are critical to the envisaged application. The present review describes the use of PGE in biosensing analysis, more specifically those sensors comprising immobilized enzymes but also briefly referring nucleic acids and other biological entities. It lays an emphasis in the immobilization process of the biological entities while focusing in the analytical performance of each biosensor, mainly sensitivity, linear range and limit of detection as comparative criteria. This review also addresses the main characteristics and properties of PGEs as transducer material in the electrochemical field.

The biocompatibility and bioactivity of hemodialysis membranes: their impact in end-stage renal disease

End-stage renal disease is a growing health problem with increasing prevalence and high health care costs. Patients suffering from end-stage renal disease exhibit higher morbidity and mortality rates compared to the general population. These patients, who are treated using hemodialysis, typically suffer from anemia, inflammation, and oxidative stress. Inadequate dialyzer membrane biocompatibility exacerbates these negative side effects. Modifications of the composition of hemodialysis membranes have improved their biocompatibility and improve the patients’ quality of life. Recently, the use of dialyzer membranes coated with bioactive compounds has also been proposed to further ameliorate dialysis-associated problems. Based on a survey of the current literature, application of bioactive membranes decreases the inflammation and oxidative stress of patients treated with hemodialysis.