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Development of a multicommutated flow system for chemiluminometric quantification of gentamicin L. Santos 1 , A. Araújo 1 , A. Fachini 1 , M. Montenegro 1 , B.F. Reis 2 1 REQUIMTE, Department of Physical Chemistry, Faculty of Pharmacy, University of Porto, R. Aníbal Cunha, 4050 Porto, Portugal. 2 Centro de Energia Nuclear na Agricultura, Universidade de S. Paulo, PO Box 96, 13400-970, Piracicaba- SP Multicommutation is a continuous flow system technique that presents several advantages relatively to flow injection analysis (FIA), such as versatility, low reagent consumption, and robustness [1]. Chemiluminescence is a high sensitivity methodology, with low detection limits that requires simple instrumentation. It is based on intensity measurements of light produced in chemiluminescent reaction. The signal intensity can be correlated with the concentration of any of the starting reagents. This analytical technique allows the quantification of a wide variety of compounds that are directly or indirectly involved in the chemiluminometric reaction [2]. The aim of this work is to quantify gentamicin in different matrices using a multicommutated flow system where the chemiluminometric oxidation of luminol by hypochlorite is accomplished. In the presence of gentamicin, an aminoglycoside antibiotic that is effective against a wide variety of gram-positive and gram-negative bacteria, the hypochlorite concentration is reduced since it oxidizes the secondary amines presents in the chemical structure of the analyte. The optimization process had been separeted in two parts: optimization of chemical conditions and optimization of flow conditions. In the first part, parameters such as the hypochlorite and luminol concentrations and the pH of the buffer solution used to prepare these reagents have being studied. After that, the flow conditions will be tested. In the final, the proposed method will be applied to the quantification of gentamicin in pharmaceutical formulations and foodstuffs. References: [1] Lavorante, A.F., Feres, M.A. and Reis, B.F. (2006), Multi-commutation in Flow Analysis: A Versatile Tool for the Development of the Automatic Analytical Procedure Focused on the Reduction of Reagent Consumption, Spectroscopy Letters, 39 (6), 631-650. [2] García-Campaña, A.M. and Baeyens, W.R.G. (2000), Principles and recent analytical applications of chemiluminescence, Analusis, 28 (8), 686-698. Acknowledgements: L. Santos thanks to Fundação Astrazeneca the master degree scholarship conceded. 137
Potentiometric determination of gentamicin in pharmaceutical formulations Martina Koci a , Célia Amorim b , Alberto Araújo b ,Maria da Conceição Montenegro b and Marie Pospíšilova, Petr Solich a Department of Analytical chemistry, Fakulty of Pharmacy, Charles University in Hradec Králové, Heyrovského 1203, Hradec Kralove 500 05, Czech Republic b REQUIMTE, Departamento de Química-Física, Faculdade de Farmácia, Universidade do Porto, Rua Aníbal Cunha, 164, 4099-030 Porto, Portugal. email: martina.koci@gmail.com Gentamicin is an aminoglycoside antibiotic, and can treat many types of bacterial infections, particularly Gram-negative infection. It is one of the heat-stable antibiotics that remain active after autoclaving, which make it particularly useful in the preparation in the certain microbiological growth media. Traditionally, pharmaceutical analysis of these compounds relies heavily on chromatography, yet also frequently requiring the use of reagent-based detection principles like spectrophotometric [1,2],chemiluminescent detection in flow-injection analysis [3], capillary electrophoresis [4] a novel LC/ELSD [5], thin-layer chromatography [6], HPLC [7].A simple and sensitive potentiometric procedure for the determination of gentamicin is presented, after development of selective electrodes based on PVC membranes using different type of cyclodextrins and its derivatives, or Evans Blue complex with gentamicin as ionophers. Several mediator solvents and lipophilic ionic species were also used in order to optimize the characteristics of the membranes. The evaluation of the general characteristics for all the units prepared revealed an analytical linear range of about 2x10 -4 ; 5x10 -3 mol L -1 , slopes of about 50 mV/dec, good reproducibility and low response times. The evaluation of Potentiometric selectivity coefficients will permit to choose the best electrode for gentamicin control in pharmaceutical preparations. References [1] El-Didamony, A. M.m Amin, A.S., Ghoneim, A.K., Telebany, A.M., (2006), Indirect spectrophotometric determination of gentamicin and vancomycin antibiotics based on their oxidation by potassium permangate, Central European J. of Chem. 4 (4), pp 708-72 [2] Ryan, J.A, (1984), Colorimetric determination of gentamicin, kanamycin, tobramycin, and amikacin aminoglycosides with 2,4-dinitrofluorobenzene, J. of Pharm. Sci. 73 (9), pp. 1301-1302 [3] Ranmos-Fernandez, J.M., Garcia-Campana, A.M.,Ales-Barrero, F.,Bosque-Sendra, (J.M, 2006), Corrigendum to “Determination of gentamicin in pharmaceutical formulations using peroxyoxalate chemiluminescent detection in flow-injection analysis, Talanta 69 (3), pp. 763-768 [4] Garcia-Ruiz, C., Marina,M.L., (2006), Recent advances in the analysis of antibiotics by capillary electrophoresis, Electrophoresis 27 (1), pp. 266-282 [5] Megoulas, N.C., Koupparis, M.A., (2004), Development and validation of a novel LC/ELSD method for the quantitation of gentamicin sulphate components in pharmaceuticals, J. Pharm. Biom. Anal. 36 (1), pp. 73- 79 [6] Sekkat, M., Fabre, H., Simeon De Bouchberg,M.m Mandrou, B., (1989), Determination of aminoglycosides in pharmaceutical formulations – I. Thin-layer chromatography, J. Pharm. Biom. Anal 7 (12), pp.883-892 [7]Fabre, H., Seddat, M., Blanchin, M.D., Mandrou, B., (1989), Determination of aminoglycosides in pharmaceutical formulations – II. High-performance liquid chromatography, J. Pharm. Biom. Anal 7(7), pp. 1711-1718 Aknowledgement: I would like to thank program Erasmus and MSMT for financial support. 138
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