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Ribeirão Preto -SP, BrasilRecebido em 1/12/06; aceito em 30/8/07; publicado na web em 26/2/08 LIQUID-PHASE MICROEXTRACTION (LPME): FUNDAMENTALS AND APPLICATIONS TO THE ANALYSIS OF DRUGS IN BIOLOGICAL SAMPLES. The analysis of drugs and metabolites in biological fluids usually requires extraction procedures to achieve sample clean-up and analyte preconcentration. Commonly, extraction procedures are performed using liquid-liquid extraction or solidphase extraction. Nevertheless, these extraction techniques are considered to be time-consuming and require a large amount of organic solvents. On this basis, microextraction techniques have been developed. Among them, liquid-phase microextraction has been standing out. This review describes the liquid-phase microextraction technique based on hollow fibers as a novel and promising alternative in sample preparation prior to chromatographic or electrophoretic analysis. The basic concepts related to this technique and its applicability in extraction of drugs are discussed.Keywords: liquid-phase microextraction; drug analysis; biological samples. INTRODUÇÃOAs técnicas de separação, tais como a cromatografia líquida de alta eficiência (HPLC), a cromatografia gasosa (GC) e a eletroforese capilar (CE), são perfeitamente adequadas para a análise de amostras contendo inúmeros componentes. No entanto, a determinação de analitos presentes em matrizes complexas, tais como plasma, soro, sangue total, homogenatos de tecidos, saliva ou urina, geralmente requer processos prévios de preparação das amostras bem elaborados. 1 Vários motivos justificam estes procedimentos, destacando-se a necessidade de eliminação prévia de parte dos componentes devido à complexidade das matrizes biológicas, a existência de proteínas que podem adsorver nas colunas cromatográficas ou nos capilares, além da necessidade de uma etapa de concentração das substâncias a serem analisadas, geralmente presentes em nível de traços. 1,2 As técnicas mais comumente utilizadas para extração e/ou pré-concentração de compostos presentes em fluidos biológicos são a extração líquido-líquido (LLE) e a extração em fase sólida (SPE). 3 Contudo, as tendências atuais apontam no sentido da utilização de menores quantidades de amostras, até mesmo para análises de traços; obtenção de maior seletividade e especificidade na extração; aumento no potencial para automação ou utilização de métodos "online", reduzindo assim a operação manual; desenvolvimento de métodos menos agressivos ao meio ambiente, com menor desperdí-cio e, o uso de quantidade mínima ou nenhuma de solventes orgâ-nicos. 4,5 Dentro deste contexto, técnicas de microextração que utilizam quantidades mínimas de solventes orgânicos e menos etapas na preparação das amostras vêm sendo desenvolvidas. Entre as principais técnicas de microextração existentes destacam-se a microextração em fase sólida (SPME), a microextração em gota suspensa (SDME) e a extração em membranas ["supported liquid membrane" (SLM), "microporous membrane liquid-liquid extraction" (MMLLE), "liquid-phase mi...

Chlorpheniramine maleate (CLOR) enantiomers were quantified by ultraviolet spectroscopy and partial least squares regression. The CLOR enantiomers were prepared as inclusion complexes with beta-cyclodextrin and 1-butanol with mole fractions in the range from 50 to 100%. For the multivariate calibration the outliers were detected and excluded and variable selection was performed by interval partial least squares and a genetic algorithm. Figures of merit showed results for accuracy of 3.63 and 2.83% (S)-CLOR for root mean square errors of calibration and prediction, respectively. The ellipse confidence region included the point for the intercept and the slope of 1 and 0, respectively. Precision and analytical sensitivity were 0.57 and 0.50% (S)-CLOR, respectively. The sensitivity, selectivity, adjustment, and signal-to-noise ratio were also determined. The model was validated by a paired t test with the results obtained by high-performance liquid chromatography proposed by the European pharmacopoeia and circular dichroism spectroscopy. The results showed there was no significant difference between the methods at the 95% confidence level, indicating that the proposed method can be used as an alternative to standard procedures for chiral analysis.

BACKGROUND The multidrug resistance (MDR) phenotype is frequently observed in Acinetobacter baumannii , the most clinically relevant pathogenic species of its genus; recently, other species belonging to the A. calcoaceticus-A. baumannii complex have emerged as important MDR nosocomial pathogens. OBJECTIVES The present study aimed to verify the occurrence of metallo-β-lactamase genes among distinct Acinetobacter species in a hospital located in the Brazilian Amazon Region. METHODS Antimicrobial susceptibility profiles were determined by broth microdilution. The genetic relationships among these isolates were assessed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Pyrosequencing reads of plasmids carrying the bla NDM-1 gene were generated using the Ion Torrent™ platform sequencing. FINDINGS A total of six isolates carried bla NDM-1 : A. baumannii (n = 2), A. nosocomialis (n = 3), and A. pittii (n = 1); three carried bla IMP-1 : A. baumannii , A. nosocomialis , and A. bereziniae . Resistance to colistin was observed for an NDM-1-producing A. nosocomialis isolate. Diverse PFGE patterns and sequence types were found among A. nosocomialis and A. baumannii isolates. The bla NDM-1 sequence was inserted in a Tn 125 transposon, while the bla IMP-1 was found as a gene cassette of the class 1 integron In 86 . MAIN CONCLUSIONS To the best of our knowledge, this is the first report describing the dissemination of bla NDM-1 among distinct Acinetobacter species recovered from the same hospital in South America.