Vu R L, Helmeste D, Albers L, Reist C
VA Medical Center, Long Beach, CA 90822, USA.
J Chromatogr B Biomed Sci Appl. 1997 Dec 5;703(1-2):195-201. doi: 10.1016/s0378-4347(97)00426-x.
A rapid and sensitive high-performance liquid chromatographic technique for simultaneous measurement of plasma venlafaxine (VEN) and its active metabolite O-desmethylvenlafaxine (ODV) is described. The process begins with the extraction of VEN and ODV, with maprotiline (MAP) as internal standard, from human plasma into an intermediate organic mixture of hexane-isoamyl alcohol and finally into an aqueous solution of 0.05% phosphoric acid. Isocratic separation of VEN, ODV and MAP is carried out by utilizing a reversed-phase butyl-bonded column (C4/E) with mobile phase consisting of acetonitrile and 40 mM phosphate buffer, pH 6.8 (50:50, v/v). Detection of VEN, ODV and MAP is done by mean of fluorimetry with excitation and emission wavelengths set at 276 and 598 nm, respectively. As low as 1.0 ng/ml VEN is detectable; while the limit of detection for ODV is 5 ng/ml. C.V. (%) of intra-day samples for both VEN and ODV are less than 10% at three concentrations tested (10.0, 50.0, 100.0 ng/ml). Similarly, over the same nominal concentrations, the precision of inter-day (5 days) samples also results in C.V. (%) smaller than 10% for both compounds, except for ODV measured at 10 ng/ml (C.V.<15%). Approximately, 100% VEN can be extracted from plasma; whereas, for ODV the recovery rate is nearly 70%. This present method is rapid, sensitive, accurate and simple for routine clinical monitoring of plasma VEN and its major metabolite ODV.
本文描述了一种快速、灵敏的高效液相色谱技术，用于同时测定血浆中的文拉法辛（VEN）及其活性代谢物O-去甲基文拉法辛（ODV）。该过程首先以马普替林（MAP）作为内标，将VEN和ODV从人血浆中萃取到己烷-异戊醇的中间有机混合物中，最后萃取到0.05%磷酸的水溶液中。VEN、ODV和MAP的等度分离是通过使用反相丁基键合柱（C4/E）进行的，流动相由乙腈和40 mM磷酸盐缓冲液（pH 6.8，50:50，v/v）组成。VEN、ODV和MAP的检测通过荧光法进行，激发波长和发射波长分别设置为276和598 nm。VEN的最低检测限为1.0 ng/ml；而ODV的检测限为5 ng/ml。在三个测试浓度（10.0、50.0、100.0 ng/ml）下，VEN和ODV日内样本的变异系数（C.V.）（%）均小于10%。同样，在相同的标称浓度下，日间（5天）样本的精密度也使得两种化合物的C.V.（%）均小于10%，但10 ng/ml的ODV测量值除外（C.V.<15%）。大约100%的VEN可以从血浆中萃取出来；而ODV的回收率接近70%。该方法快速、灵敏、准确且简单，适用于血浆VEN及其主要代谢物ODV的常规临床监测。
