Department of Chemistry, Amirkabir University of Technology, Tehran, Iran.
Talanta. 2009 Nov 15;80(1):133-8. doi: 10.1016/j.talanta.2009.06.033. Epub 2009 Jun 21.
In this work, a novel method is described for the determination of bromhexine in biological fluids using molecularly imprinted solid-phase extraction as the sample cleanup technique combined with high performance liquid chromatography (HPLC). The water-compatible molecularly imprinted polymers (MIPs) were prepared using methacrylic acid as functional monomer, ethylene glycol dimethacrylate as cross-linker, chloroform as porogen and bromhexine as the template molecule. The novel imprinted polymer was used as a solid-phase extraction sorbent for the extraction of bromhexine from human serum and urine. Various parameters affecting the extraction efficiency of the polymer have been evaluated. The optimal conditions for molecularly imprinted solid-phase extraction (MISPE) consisted of conditioning 1 mL methanol and 1 mL of deionized water at neutral pH, loading of 5 mL of the water sample (25 microg L(-1)) at pH 6.0, washing using 2 mL acetonitrile/acetone (1/4, v/v) and elution with 3x 1 mL methanol/acetic acid (10/1, v/v). The MIP selectivity was evaluated by checking several substances with similar molecular structures to that of bromhexine. Results from the HPLC analyses showed that the calibration curve of bromhexine using MIP from human serum and urine is linear in the ranges of 0.5-100 and 1.5-100 microg L(-1) with good precisions (3.3% and 2.8% for 5.0 microg L(-1)), respectively. The recoveries for serum and urine samples were higher than 92%.
在这项工作中，描述了一种使用分子印迹固相萃取（作为样品净化技术）结合高效液相色谱（HPLC）测定生物体液中盐酸氨溴索的新方法。采用甲基丙烯酸作为功能单体、乙二醇二甲基丙烯酸酯作为交联剂、氯仿作为致孔剂、盐酸氨溴索作为模板分子，制备了亲水性分子印迹聚合物（MIPs）。将新型印迹聚合物用作固相萃取吸附剂，从人血清和尿液中提取盐酸氨溴索。评价了影响聚合物萃取效率的各种参数。分子印迹固相萃取（MISPE）的最佳条件包括：用中性 pH 值的 1 mL 甲醇和 1 mL 去离子水预处理，用 pH 值为 6.0 的 5 mL 水样（25 μg L(-1))）加载，用 2 mL 乙腈/丙酮（1/4，v/v）洗涤，用 3x 1 mL 甲醇/乙酸（10/1，v/v）洗脱。通过检查几种与盐酸氨溴索具有相似分子结构的物质来评估 MIP 的选择性。HPLC 分析结果表明，用人血清和尿液中的 MIP 测定盐酸氨溴索的校准曲线在 0.5-100 和 1.5-100 μg L(-1) 范围内呈线性，具有良好的精密度（5.0 μg L(-1) 时分别为 3.3%和 2.8%）。血清和尿液样品的回收率均高于 92%。
