da Silva Anne Caroline Cezimbra, Raasch Juliana Raquel, Vargas Tainara Gomes, Peteffi Giovana Piva, Hahn Roberta Zilles, Antunes Marina Venzon, Perassolo Magda Susana, Linden Rafael
Laboratory of Analytical Toxicology, Instituto de Ciências da Saúde, Universidade Feevale, Novo Hamburgo, RS, Brazil.
Laboratory of Analytical Toxicology, Instituto de Ciências da Saúde, Universidade Feevale, Novo Hamburgo, RS, Brazil.
Clin Biochem. 2018 Feb;52:85-93. doi: 10.1016/j.clinbiochem.2017.10.002. Epub 2017 Oct 5.
Therapeutic drug monitoring (TDM) of the widely prescribed antidepressant fluoxetine (FLU) is recommended in certain situations, such as occurrence of toxicity, inadequate response or suspect of poor adherence. Dried blood spot (DBS) sampling is an increasingly studied alternative for TDM, particularly for outpatients, due to its ease of collection and inherent stability.
The aim of this study was to develop and validate an LC-MS/MS assay for the simultaneous quantification of FLU and norfluoxetine (NFLU) in DBS.
The assay is based on a liquid extraction of single DBS with 8mm of diameter, using FLU-D6 as the internal standard, followed by reversed phase separation in an Accucore® C18 column (100×2.1mm, 2.6μm). Mobile phase was composed of water and acetonitrile (gradient from 80:20 to 50:50, v/v), both containing formic acid 0.1%. The assay was validated and applied to 30 patients under FLU pharmacotherapy.
The assay was linear in the range 10-750ngmL. Precision assays presented CV% of 3.13-9.61 and 3.54-7.99 for FLU and NFLU, respectively, and accuracy in the range of 97.98-110.44% and 100.25-105.8%. FLU and NFLU were stable at 25 and 45°C for 7days. The assay was evaluated in 30 patients under FLU treatment. Concentrations of both compounds were higher in DBS than in plasma, and the use of the multiplying factors 0.71 and 0.68 for FLU and NFLU, respectively, allowed acceptable estimation of plasma concentrations, with median prediction bias of -0.55 to 0.55% and mean differences of 0.4 to 2.2ngmL.
The presented data support the clinical use of DBS for therapeutic drug monitoring of FLU.
在某些情况下，如出现毒性反应、疗效不佳或怀疑依从性差时，建议对广泛使用的抗抑郁药氟西汀（FLU）进行治疗药物监测（TDM）。由于易于采集且具有内在稳定性，干血斑（DBS）采样作为TDM的一种替代方法，正受到越来越多的研究，特别是对于门诊患者。
本研究的目的是开发并验证一种用于同时定量DBS中FLU和去甲氟西汀（NFLU）的液相色谱-串联质谱（LC-MS/MS）测定法。
该测定法基于用8毫米直径的单个DBS进行液液萃取，以氟西汀-D6作为内标，然后在Accucore® C18柱（100×2.1毫米，2.6微米）中进行反相分离。流动相由水和乙腈组成（梯度从80:20至50:50，v/v），两者均含有0.1%的甲酸。该测定法经过验证，并应用于30例接受氟西汀药物治疗的患者。
该测定法在10 - 750纳克/毫升范围内呈线性。精密度测定中，FLU和NFLU的CV%分别为3.13 - 9.61和3.54 - 7.99，准确度范围分别为97.98 - 110.44%和100.25 - 105.8%。FLU和NFLU在25℃和45℃下可稳定保存7天。对30例接受氟西汀治疗的患者进行了该测定法的评估。两种化合物在DBS中的浓度均高于血浆中的浓度，分别使用0.71和0.68的换算因子用于FLU和NFLU，可以对血浆浓度进行可接受的估算，中位预测偏差为 - 0.55至0.55%，平均差异为0.4至2.2纳克/毫升。
所提供的数据支持将DBS用于氟西汀的治疗药物监测的临床应用。
