Departamento de Ciências Farmacêuticas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
Electrophoresis. 2011 Oct;32(19):2765-75. doi: 10.1002/elps.201100328. Epub 2011 Sep 5.
Knowing that microbial transformations of compounds play vital roles in the preparation of new derivatives with biological activities, risperidone and its chiral metabolites were determined by capillary electrophoresis and hollow fiber liquid-phase microextraction after a fungal biotransformation study in liquid culture medium. The analytes were extracted from 1 mL liquid culture medium 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 100 mmol/L sodium phosphate buffer pH 3.0 containing 2.0% w/v sulfated-α-CD and carboxymethyl-β-CD 0.5% w/v with a constant voltage of -10 kV. The method was linear over the concentration range of 100-5000 ng/mL for risperidone and 50-5000 ng/mL for each metabolite enantiomer. Within-day and between-day assay precisions and accuracies for all the analytes were studied at three concentration levels, and the values of relative standard deviation and relative error were lower than 15%. The developed method was applied in a pilot biotransformation study employing risperidone as the substrate and the filamentous fungus Mucor rouxii. This study showed that the filamentous fungus was able to metabolize risperidone enantioselectively into its chiral active metabolite, (-)-9-hydroxyrisperidone.
鉴于微生物对化合物的转化在制备具有生物活性的新衍生物方面起着至关重要的作用，在液体培养基中的真菌生物转化研究之后，通过毛细管电泳和中空纤维液相微萃取来确定利培酮及其手性代谢物。将分析物从 1 mL 液体培养基中提取到中空纤维孔中的 1-辛醇中，并提取到聚丙稀中空纤维内部的酸接受溶液中。在含有 2.0%w/v 硫酸化-α-CD 和 0.5%w/v 羧甲基-β-CD 的 100mmol/L 磷酸钠缓冲液 pH3.0 中，在-10 kV 的恒电压下进行电泳分离。该方法在利培酮的浓度范围为 100-5000ng/mL 和每个代谢物对映体的 50-5000ng/mL 范围内呈线性。在三个浓度水平下研究了所有分析物的日内和日间测定精密度和准确度，相对标准偏差和相对误差的值低于 15%。该方法已应用于利培酮作为底物的丝状真菌毛霉的初步生物转化研究。该研究表明，丝状真菌能够对映选择性地将利培酮代谢为其手性活性代谢物(-)-9-羟基利培酮。
