Darwish Ibrahim A, Al-Majed Abdulrahman A, Mahmoud Ashraf M, Khalil Nasr Y
King Saud University, College of Pharmacy, Department of Pharmaceutical Chemistry, PO Box 2457, Riyadh 11451, Saudi Arabia.
J AOAC Int. 2009 Sep-Oct;92(5):1349-55.
New nonextractive and simple offline precolumn derivatization procedures have been proposed, for the first time, for the trace determination of paroxetine (PXT) in human plasma by HPLC with fluorescence detection. Trimetazidine (TMZ) was used as an internal standard. Plasma samples were treated with acetonitrile for protein precipitation and then derivatized with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole in borate buffer of pH 8 at 70 degrees C for 30 min. Separations of the derivatized PXT and TMZ were performed on a Nucleosil CN column using a mobile phase consisting of acetonitrile-10 mM sodium acetate buffer (pH 3.5)-methanol (47 + 47 + 6, v/v) at a flow rate of 1.0 mL/min. The derivatized samples were excited at 470 nm and monitored at an emission wavelength of 530 nm. Under the optimum chromatographic conditions, a linear relationship with good correlation coefficient (r = 0.9998, n = 7) was found between the peak area ratio and PXT concentrations in the range of 5-600 ng/mL. The LOD and LOQ were 1.37 and 4.14 ng/mL, respectively. The intraday and interassay precisions were satisfactory; the RSD did not exceed 4.2%. The accuracy of the method was proved by recovery of PXT from spiked human plasma at levels of 97.28-104.38 +/- 0.41-3.62%. The proposed method had high throughput, as the analysis involved a simple sample pretreatment procedure and short run time (< 10 min). The results demonstrated that the method would have a great value when it is applied in the therapeutic monitoring of PXT.
首次提出了新型非萃取且简单的离线柱前衍生化程序，用于通过高效液相色谱-荧光检测法痕量测定人血浆中的帕罗西汀（PXT）。曲美他嗪（TMZ）用作内标。血浆样品用乙腈进行蛋白沉淀处理，然后在70℃下于pH 8的硼酸盐缓冲液中用7-氯-4-硝基苯并-2-恶唑-1,3-二唑衍生化30分钟。衍生化的PXT和TMZ在Nucleosil CN柱上进行分离，流动相由乙腈-10 mM醋酸钠缓冲液（pH 3.5）-甲醇（47 + 47 + 6，v/v）组成，流速为1.0 mL/min。衍生化样品在470 nm处激发，在发射波长530 nm处监测。在最佳色谱条件下，在5-600 ng/mL范围内，峰面积比与PXT浓度之间发现了具有良好相关系数（r = 0.9998，n = 7）的线性关系。检测限和定量限分别为1.37和4.14 ng/mL。日内和日间精密度令人满意；相对标准偏差不超过4.2%。通过在加标人血浆中PXT的回收率为97.28-104.38 +/- 0.41-3.62%，证明了该方法的准确性。所提出的方法具有高通量，因为分析涉及简单的样品预处理程序和较短的运行时间（<10分钟）。结果表明，该方法应用于PXT的治疗监测时具有很大价值。
