Magalhães Paulo, Alves Gilberto, Llerena Adrián, Falcão Amílcar
Laboratory of Pharmacology, Faculty of Pharmacy, University of Coimbra, Pólo das Ciências da Saúde, Azinhaga de Santa Comba, 3000-548 Coimbra, Portugal.
CNC-Center for Neuroscience and Cell Biology, University of Coimbra, 3004-517 Coimbra, Portugal.
J Anal Toxicol. 2017 Sep 1;41(7):631-638. doi: 10.1093/jat/bkx043.
The present article reports the first liquid chromatography (LC) assay for the simultaneous quantification of fluoxetine (FLU), its metabolite norfluoxetine (NFLU) and paroxetine (PAR) in human plasma, applying the microextraction by packed sorbent (MEPS) technology in sample preparation. Chromatographic analysis of FLU, NFLU and PAR was achieved in <13 min on a reverse-phase C18 column using isocratic elution and fluorescence detection (FLD). The mobile phase was composed by an aqueous solution of 25 mM sodium phosphate monobasic anhydrous and 7.5 mM di-potassium hydrogen phosphate anhydrous (pH 3.0)/acetonitrile/methanol (70:23:7, v/v/v). The detector was set at 240/312 nm for FLU, NFLU and IS and at 295/350 nm for PAR. The method showed linearity in the ranges of 20-750 ng mL-1 for FLU and NFLU, and 5-750 ng mL-1 for PAR (r2 ≥ 0.9919). The overall intra- and interday precision did not exceed 13.6% and the corresponding accuracy (bias) ranged from 0.02 to 16.7%. The method was successfully applied in the analysis of authentic plasma samples. Hence, this new MEPS/LC-FLD assay ensures robust and low-cost analyses representing, therefore, a good alternative to support therapeutic drug monitoring and clinical studies involving these antidepressant drugs.
本文报道了首个用于同时定量测定人血浆中氟西汀（FLU）、其代谢产物去甲氟西汀（NFLU）和帕罗西汀（PAR）的液相色谱（LC）分析方法，该方法在样品制备中采用了填充吸附剂微萃取（MEPS）技术。使用等度洗脱和荧光检测（FLD），在反相C18柱上于<13分钟内完成了FLU、NFLU和PAR的色谱分析。流动相由25 mM无水磷酸二氢钠和7.5 mM无水磷酸氢二钾的水溶液（pH 3.0）/乙腈/甲醇（70:23:7，v/v/v）组成。FLU、NFLU和内标物（IS）的检测波长设置为240/312 nm，PAR的检测波长设置为295/350 nm。该方法在FLU和NFLU的20 - 750 ng mL-1范围内以及PAR的5 - 750 ng mL-1范围内呈线性（r2≥0.9919）。总体日内和日间精密度不超过13.6%，相应的准确度（偏差）范围为0.02%至16.7%。该方法成功应用于实际血浆样品的分析。因此，这种新的MEPS/LC - FLD分析方法确保了稳健且低成本的分析，从而为支持涉及这些抗抑郁药物的治疗药物监测和临床研究提供了一个很好的替代方案。
