Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, 14040-901, Ribeirão Preto, SP, Brazil.
J Pharm Biomed Anal. 2012 Mar 5;61:100-7. doi: 10.1016/j.jpba.2011.12.012. Epub 2011 Dec 21.
A high-performance liquid chromatographic method using polar organic mode was developed to analyze albendazole (ABZ), albendazole sulfone (ABZSO(2)) and the chiral and active metabolite albendazole sulfoxide (ABZSOX, ricobendazole) that was further applied in stereoselective fungal biotransformation studies. The chromatographic separation was performed on a Chiralpak AS column using acetonitrile:ethanol (97:3, v/v) plus 0.2% triethylamine and 0.2% acetic acid as the mobile phase at a flow rate of 0.5 mL min(-1). The present study employed hollow fiber liquid-phase microextraction as sample preparation. The method showed to be linear over the concentration range of 25-5000 ng mL(-1) for each ABZSOX enantiomer, 200-10,000 ng mL(-1) for ABZ and 50-1000 ng mL(-1) for ABZSO(2) metabolite with correlation coefficient (r)>0.9934. The mean recoveries for ABZ, rac-ABZSOX and ABZSO(2) were, respectively, 9%, 33% and 20% with relative standard deviation below 10%. Within-day and between-day precision and accuracy assays for these analytes were studied at three concentration levels and were lower than 15%. This study opens the door regarding the possibility of using fungi in obtaining of the active metabolite ricobendazole. Nigrospora sphaerica (Sacc.) E. W. Mason (SS67), Pestalotiopsis foedans (VR8), Papulaspora immersa Hotson (SS13) and Mucor rouxii were able to stereoselectively metabolize ABZ into its chiral metabolite. Among them, the fungus Mucor rouxii was the most efficient in the production of (+)-ABZSOX.
建立了一种采用极性有机模式的高效液相色谱法，用于分析阿苯达唑（ABZ）、阿苯达唑砜（ABZSO(2)）和手性活性代谢物阿苯达唑砜（ABZSOX，瑞波西隆），并进一步应用于立体选择性真菌生物转化研究。在 Chiralpak AS 柱上，以乙腈：乙醇（97:3，v/v）加 0.2%三乙胺和 0.2%乙酸作为流动相，流速为 0.5 mL min(-1)，进行色谱分离。本研究采用中空纤维液相微萃取作为样品制备方法。该方法对每个 ABZSOX 对映体的浓度范围为 25-5000ng mL(-1)、ABZ 的浓度范围为 200-10000ng mL(-1)和 ABZSO(2)代谢物的浓度范围为 50-1000ng mL(-1)时表现出线性关系，相关系数(r)>0.9934。ABZ、rac-ABZSOX 和 ABZSO(2)的平均回收率分别为 9%、33%和 20%，相对标准偏差低于 10%。在三个浓度水平下研究了这些分析物的日内和日间精密度和准确度，结果均低于 15%。这项研究为利用真菌获得活性代谢物瑞波西隆开辟了可能性。黑曲霉（Sacc.）E. W. Mason（SS67）、拟青霉（VR8）、浸没曲霉（SS13）和毛霉（Mucor rouxii）能够立体选择性地将 ABZ 代谢为其手性代谢物。其中，毛霉（Mucor rouxii）在生产（+）-ABZSOX 方面最为有效。
