Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, São Paulo, Brazil.
Electrophoresis. 2011 Oct;32(19):2746-56. doi: 10.1002/elps.201000658. Epub 2011 Sep 8.
Hollow fiber liquid-phase microextraction and CE were applied for the determination of albendazole sulfoxide (ASOX) enantiomers in liquid culture medium after a fungal biotransformation study. The analytes were extracted from 1 mL of liquid culture medium spiked with the internal standard (rac-hydroxychloroquine) and buffered with 0.50 mol/L phosphate buffer, pH 10. The analytes were extracted into 1-octanol impregnated in the pores of the hollow fiber, and into an acid acceptor solution inside the polypropylene hollow fiber. The electrophoretic separations were carried out in 0.05 mol/L tris(hydroxymethyl)aminomethane buffer, pH 9.3, containing 3.0% w/v sulfated-β-CD (S-β-CD) with a constant voltage of +15 kV and detection at 220 nm. The method was linear over the concentration range of 250-5000 ng/mL for each ASOX enantiomer. Within-day and between-day assay precision and accuracy for the analytes were studied at three concentration levels and the values of RSD% and relative error % were lower than 15%. The developed method was applied for the determination of ASOX after a biotransformation study employing the endophytic fungus Penicillium crustosum (VR4). This study showed that the endophytic fungus was able to metabolize the albendazole to ASOX enantioselectively. In addition, it was demonstrated that hollow fiber liquid-phase microextraction coupled to CE can be an excellent and environmentally friendly technique for the analysis of samples obtained in biotransformation studies.
采用中空纤维液相微萃取和毛细管电泳法测定真菌生物转化后液体培养基中阿苯达唑砜(ASOX)对映体。用内标(rac-羟氯喹)和 0.50mol/L 磷酸盐缓冲液(pH10)缓冲的 1ml 液体培养基中提取分析物。分析物被萃取到中空纤维孔内的 1-辛醇中,并被萃取到聚丙烯中空纤维内的酸接受溶液中。电泳分离在 0.05mol/L 三羟甲基氨基甲烷缓冲液(pH9.3)中进行,其中含有 3.0%w/v 硫酸化-β-环糊精(S-β-CD),外加 15kV 的恒定电压,在 220nm 处检测。对于每个 ASOX 对映体,该方法在 250-5000ng/mL 的浓度范围内呈线性。在三个浓度水平下研究了分析物的日内和日间测定精密度和准确度,RSD%和相对误差%的值低于 15%。该方法应用于内生真菌 Penicillium crustosum(VR4)进行生物转化研究后 ASOX 的测定。该研究表明,内生真菌能够将阿苯达唑选择性地代谢为 ASOX 对映体。此外,还证明了中空纤维液相微萃取与毛细管电泳的结合可以作为生物转化研究中获得的样品分析的一种极好的环保技术。