A simple and reliable method for quantifying plasma concentrations of tetracyclic antidepressants using monolithic silica solid-phase extraction tips
A simple and reliable method for quantifying plasma concentrations of tetracyclic antidepressants using monolithic silica solid-phase extraction tips
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DOI：
10.1007/s11419-012-0132-y
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发表时间：
2012-03
影响因子：
2.2
通讯作者：
Daigo Hayashi;T. Kumazawa;Chika Hasegawa;X. Lee;A. Marumo;S. Uchigasaki;Mitsuru Kawamura;Keizo Sato
中科院分区：
文献类型：
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作者：
Daigo Hayashi;T. Kumazawa;Chika Hasegawa;X. Lee;A. Marumo;S. Uchigasaki;Mitsuru Kawamura;Keizo Sato
使用含有整体硅胶填充材料 MonoTip C18 的新型微量移液器固相萃取头从人血浆样品中提取三种四环抗抑郁药米安色林、米氮平和司替替林。将含有米安舍林、米氮平和司替替林的血浆样品 (0.1 ml) 与 330 μl 蒸馏水和 50 μl 0.5 M 甘氨酸-氢氧化钠缓冲溶液 (pH 8) 混合。离心后，通过 25 个连续的吸液/分配循环将上清液提取到吸头的 C18 相上（移液器吸头体积，200 μl）。通过 5 个吸取/分配循环，用甲醇洗脱保留在 C18 相中的三种分析物。洗脱液通过气相色谱 (GC) 和氮磷检测 (NPD) 进行测量。三种抗抑郁药的回收率为 84.6–99.6%，每种药物的检测限在 2.4 至 15 ng/ml 血浆之间。药物的最大日内和日间变异系数低于10.6%。三种药物的回归方程在米安舍林和米氮平的 50–5000 ng/ml 范围内以及司替替林的 50–10 000 ng/ml 范围内表现出良好的线性。抗抑郁药在 4°C 下稳定至少 12 小时，在 -80°C 下稳定至少 4 周，并在血浆中经过 3 个冻融循环。经过验证的方法已成功用于定量人类受试者口服药物后米氮平的血浆浓度。虽然我们用 GC-NPD 实现了分析物检测，但目前的四环抗抑郁药整体微量移液器萃取方法似乎非常有助于将其与 GC-质谱 (MS) 和/或液相色谱-串联 MS 相结合，以确保高度的简单性、纯化性、重现性和灵敏度。
Three tetracyclic antidepressants, mianserin, mirtazapine, and setiptiline, were extracted from human plasma samples using a new micropipette solid-phase extraction tip containing a monolithic silica gel packing material, the MonoTip C18. The plasma samples (0.1 ml), which contained mianserin, mirtazapine, and setiptiline, were mixed with 330 μl of distilled water and 50 μl of 0.5 M glycine–sodium hydroxide buffer solution (pH 8). After centrifugation, the supernatant was extracted onto the C18phase of the tip (pipette tip volume, 200 μl) by 25 sequential aspirating/dispensing cycles. The three analytes retained in the C18phase were eluted with methanol by 5 aspirating/dispensing cycles. The eluate was subjected to measurements by gas chromatography (GC) with nitrogen–phosphorus detection (NPD). The recoveries of the three antidepressants were 84.6–99.6% and the limits of detection for each drug were between 2.4 and 15 ng/ml of plasma. The maximum intraday and interday coefficients of variation for the drugs was below 10.6%. Regression equations for the three drugs showed excellent linearity in the range of 50–5000 ng/ml for mianserin and mirtazapine and 50–10 000 ng/ml for setiptiline. The antidepressants were stable for at least 12 h at 4°C, 4 weeks at −80°C, and through three freeze–thaw cycles in plasma. The validated method was successfully used to quantify the plasma concentrations of mirtazapine in a human subject after oral administration of the drug. Although we achieved analyte detection with GC-NPD, the present monolithic micropipette extraction method for tetracyclic antidepressants seems to be very useful for combining it with GC–mass spectrometry (MS) and/or liquid chromatography–tandem MS to secure high simplicity, purification, reproducibility, and sensitivity.