A rapid, sensitive, selective and simple capillary zone electrophoresis (CZE) method, by integration of field amplified sample stacking (FASS) and in-capillary derivatization (in-CapD) coupled with fluorescence (FL) detection (CZE-FASS-in-CapD-FL), has been developed and validated for simultaneous determination of morphine and its metabolite, morphine-6-glucuronide (morphine-6-G)in human urine. The method was based on in-line derivatization by introducing the sample to running background electrolytes (BGE) of disodium tetraborate decahydrate and potassium ferricyanide, whereas analytes were oxidized into highly fluorescent dimer products. Various parameters influencing the sensitivity and efficiency of the proposed method were investigated, and the optimal conditions of the BGE were as follow: 60 mM disodium tetraborate decahydrate (pH 10.5), 0.25 mM potassium ferricyanide and separation voltage of 9 kV. For FASS, morphine and morphine-6-G were electrokinetically injected for 14 s at 16 kV into the capillary that was pre-injected with water plug for 4 s. The fluorescence signals were then detected at excitation and emission wavelengths of 340 and 450 nm, respectively. Analysis was performed at ambient temperature (22±1 °C). Validation of the method showed good linearity over morphine and morphine-6-G concentrations ranging from 2 to 2000 ng/mL and 2.5 to 2000 ng/mL, respectively. The mean recoveries were ranged from 97.72 to 87.26% and 97.14 to 85.43% for morphine and morphine-6-G, respectively. The validated CZE-FASS-in-CapD-FL method was effectively applied as quantitative forensic analysis of morphine and morphine-6-G in clinical urine samples.

一种快速，灵敏，选择性和简单的毛细管区带电泳（CZE）方法，方法是将现场扩增的样品堆叠（FASS）和毛细管内衍生化（in-CapD）与荧光（FL）检测（CZE-FASS-in- CapD-FL）已开发并验证可用于同时测定人尿液中的吗啡及其代谢物吗啡-6-葡萄糖醛酸（吗啡-6-G）。该方法基于在线衍生化，方法是将样品引入十水合四硼酸钠和铁氰化钾的运行本底电解质（BGE）中，而分析物被氧化成高荧光度的二聚体产物。研究了影响该方法灵敏度和效率的各种参数，BGE的最佳条件如下：60 mM十水合四硼酸钠（pH 10.5），0。25 mM铁氰化钾，分离电压为9 kV。对于FASS，将吗啡和吗啡6-G在16 kV下电动注入毛细管14 s，该毛细管已预先注有水塞4 s。然后分别在340和450nm的激发和发射波长处检测荧光信号。分析在环境温度（22±1°C）下进行。该方法的验证显示，吗啡和吗啡-6-G浓度分别在2至2000 ng / mL和2.5至2000 ng / mL的浓度范围内具有良好的线性。吗啡和吗啡-6-G的平均回收率分别为97.72％至87.26％和97.14％至85.43％。经过验证的CZE-FASS-in-CapD-FL方法有效地用于临床尿液样品中吗啡和吗啡-6-G的定量法医分析。