Department of Chemistry, Amirkabir University of Technology, Tehran, Iran.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Dec 12;911:49-54. doi: 10.1016/j.jchromb.2012.10.019. Epub 2012 Oct 23.
The applicability of an on-line solid phase extraction method using molecularly imprinted monolithic column was developed for the assay of tramadol (TRD) in urine and plasma samples. The monolithic column was prepared by using TRD as the template, methacrylic acid (MAA) as the functional monomer, ethylene glycol dimethacrylate (EGDMA) as the cross-linker and chloroform as the porogen with in situ molecular imprinting polymerization technique. Various parameters affecting the extraction efficiency of the monolithic column were evaluated. Chromatographic analysis of TRD after on-line clean-up of samples was performed by reversed-phase HPLC on an ACE column with ultraviolet detection at 218nm. The present work was successfully applied for automated simple analysis of TRD in urine and plasma samples with high recoveries between 90.5-93.1% and 93.3-96.0%, respectively. The results revealed that in concentration up to 500ng/mL of dextromethorphan (DEX), timolol (TMO) and O-desmethyltramadol (M1), the recoveries were not reduced more than 4.3% and 4.0% for plasma and urine samples, respectively. The limit of detection (S/N=3) and limit of quantification (S/N=10) for TRD in urine samples were 0.03ng/mL and 0.10ng/mL, and in plasma samples were 0.3 and 1.0ng/mL, respectively. Inter-column precision of the assays (n=3) for urine and plasma samples at the 100ng/mL TRD level were 4.0% and 4.2%, respectively.
采用分子印迹整体柱的在线固相萃取法被开发用于尿液和血浆样品中曲马多（TRD）的测定。整体柱是通过以 TRD 为模板、甲基丙烯酸（MAA）为功能单体、乙二醇二甲基丙烯酸酯（EGDMA）为交联剂和氯仿为致孔剂，采用原位分子印迹聚合技术制备的。评估了影响整体柱萃取效率的各种参数。样品在线净化后，通过反相 HPLC 在 ACE 柱上进行色谱分析，在 218nm 处进行紫外检测。本工作成功地应用于尿液和血浆样品中 TRD 的自动化简单分析，回收率分别为 90.5-93.1%和 93.3-96.0%。结果表明，在去甲右美沙芬（DEX）、噻吗洛尔（TMO）和 O-去甲曲马多（M1）浓度高达 500ng/mL 的情况下，血浆和尿液样品的回收率均未降低超过 4.3%和 4.0%。尿液样品中 TRD 的检测限（S/N=3）和定量限（S/N=10）分别为 0.03ng/mL 和 0.10ng/mL，血浆样品分别为 0.3 和 1.0ng/mL。尿液和血浆样品中 100ng/mL TRD 水平的柱间精密度（n=3）分别为 4.0%和 4.2%。
