Laboratory of Analytical Chemistry, Department of Chemistry, University of Ioannina, Ioannina, Greece.
J Sep Sci. 2013 Jun;36(11):1720-5. doi: 10.1002/jssc.201300106. Epub 2013 May 3.
In the present study, we propose the first HPLC method coupled to postcolumn derivatization for the determination of rimantadine in human urine samples. The analyte and amantadine (internal standard) were isocratically separated using an RP monolithic stationary phase (100 × 4.6 mm id) with a mobile phase consisting of CH3OH/phosphate buffer (25 mmol/L, pH 3.0) at a volume ratio of 50:50. Postcolumn derivatization involved on-line reaction with o-phthalaldehyde (20 mmol/L) and N-acetyl-cysteine (5 mmol/L) at alkaline medium (100 mmol/L borate pH 11.0). Spectrofluorimetric detection at λ(ex)/λ(em) = 340/455 nm enabled the selective and sensitive determination of rimantadine in urine samples at a range of 50-500 ng/mL with an LOD of 5 ng/mL. Human urine samples were analyzed successfully after SPE using hydrophilic-lipophilic balanced RP cartridges (30 mg/mL, Oasis HLB). Recoveries ranged between 89.7 and 102.7%.
在本研究中，我们提出了第一个 HPLC 方法，结合柱后衍生化，用于测定人尿样中的金刚烷胺。采用 RP 整体固定相（100×4.6mm id），以甲醇/磷酸盐缓冲液（25mmol/L，pH3.0）为流动相，体积比为 50:50，对分析物和金刚烷胺（内标）进行等度分离。柱后衍生化涉及在线与邻苯二醛（20mmol/L）和 N-乙酰半胱氨酸（5mmol/L）在碱性介质（100mmol/L 硼酸盐 pH11.0）中的反应。在 λ（ex）/λ（em）=340/455nm 处的荧光检测，使金刚烷胺在 50-500ng/mL 的范围内得到选择性和灵敏的测定，LOD 为 5ng/mL。使用亲水-疏水平衡 RP 小柱（30mg/mL，OasisHLB）进行 SPE 后，成功地分析了人尿样。回收率在 89.7%至 102.7%之间。
