Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Hradec Králové, Czech Republic.
Anal Bioanal Chem. 2013 Aug;405(20):6583-7. doi: 10.1007/s00216-013-7098-4. Epub 2013 Jun 11.
A new and fast high-performance liquid chromatography (HPLC) column-switching method using fused-core columns in both dimensions for sample preconcentration and determination of propranolol in human urine has been developed. On-line sample pretreatment and propranolol preconcentration were performed on an Ascentis Express RP-C-18 guard column (5 × 4.6 mm), particle size, 2.7 μm, with mobile phase acetonitrile/water (5:95, v/v) at a flow rate of 2.0 mL min(-1) and at a temperature of 50 °C. Valve switch from pretreatment column to analytical column was set at 4.0 min in a back-flush mode. Separation of propranolol from other endogenous urine compounds was achieved on the fused-core column Ascentis Express RP-Amide (100 × 4.6 mm), particle size, 2.7 μm, with mobile phase acetonitrile/water solution of 0.5% triethylamine, pH adjusted to 4.5 by means of glacial acetic acid (25:75, v/v), at a flow rate of 1.0 mL min(-1) and at a temperature of 50 °C. Fluorescence excitation/emission detection wavelengths were set at 229/338 nm. A volume of 1,500 μL of filtered urine sample solution was injected directly into the column-switching HPLC system. The total analysis time including on-line sample pretreatment was less than 8 min. The experimentally determined limit of detection of the method was found to be 0.015 ng mL(-1).
已开发出一种新的快速高效液相色谱（HPLC）柱切换方法，该方法在两个维度上均使用熔融核柱进行样品预浓缩和测定人尿中的普萘洛尔。在线样品预处理和普萘洛尔预浓缩在 Ascentis Express RP-C-18 保护柱（5×4.6mm）上进行，粒径为 2.7μm，流动相为乙腈/水（5：95，v/v），流速为 2.0mL min（-1），温度为 50°C。在反冲洗模式下，将阀从预处理柱切换到分析柱的时间设置为 4.0min。在熔融核柱 Ascentis Express RP-Amide（100×4.6mm）上，粒径为 2.7μm，从其他内源性尿液化合物中分离出普萘洛尔，采用乙腈/水的 0.5%三乙胺溶液，通过冰醋酸将 pH 值调节至 4.5（25：75，v/v），流速为 1.0mL min（-1），温度为 50°C。荧光激发/发射检测波长设置为 229/338nm。直接将 1500μL 过滤的尿液样品溶液注入柱切换 HPLC 系统。包括在线样品预处理在内的总分析时间小于 8 分钟。该方法的实验确定检测限为 0.015ng mL（-1）。
