Wang Li, Zhang Zhujun, Huang Lianggao
School of Chemistry and Materials Science, Shaanxi Normal University, Xi'an, 710062, China.
Anal Bioanal Chem. 2008 Mar;390(5):1431-6. doi: 10.1007/s00216-007-1795-9. Epub 2008 Jan 8.
A new molecularly imprinted polymer (MIP)-chemiluminescence (CL) imaging detection approach towards chiral recognition of dansyl-phenylalanine (Phe) is presented. The polymer microspheres were synthesized using precipitation polymerization with dansyl-L-Phe as template. Polymer microspheres were immobilized in microtiter plates (96 wells) using poly(vinyl alcohol) (PVA) as glue. The analyte was selectively adsorbed on the MIP microspheres. After washing, the bound fraction was quantified based on peroxyoxalate chemiluminescence (PO-CL) analysis. In the presence of dansyl-Phe, bis(2,4,6-trichlorophenyl)oxalate (TCPO) reacted with hydrogen peroxide (H2O2) to emit chemiluminescence. The signal was detected and quantified with a highly sensitive cooled charge-coupled device (CCD). Influencing factors were investigated and optimized in detail. Control experiments using capillary electrophoresis showed that there was no significant difference between the proposed method and the control method at a confidence level of 95%. The method can perform 96 independent measurements simultaneously in 30 min and the limits of detection (LODs) for dansyl-L-Phe and dansyl-D-Phe were 0.025 micromol L(-1) and 0.075 micromol L(-1) (3sigma), respectively. The relative standard deviation (RSD) for 11 parallel measurements of dansyl-L-Phe (0.78 micromol L(-1)) was 8%. The results show that MIP-based CL imaging can become a useful analytical technology for quick chiral recognition.
本文提出了一种用于手性识别丹磺酰苯丙氨酸（Phe）的新型分子印迹聚合物（MIP）-化学发光（CL）成像检测方法。以丹磺酰-L-苯丙氨酸为模板，通过沉淀聚合法合成了聚合物微球。使用聚乙烯醇（PVA）作为胶水将聚合物微球固定在微孔板（96孔）中。分析物被选择性地吸附在MIP微球上。洗涤后，基于过氧草酸酯化学发光（PO-CL）分析对结合部分进行定量。在丹磺酰-Phe存在的情况下，双（2,4,6-三氯苯基）草酸酯（TCPO）与过氧化氢（H2O2）反应发出化学发光。信号通过高灵敏度的冷却电荷耦合器件（CCD）进行检测和定量。详细研究并优化了影响因素。使用毛细管电泳的对照实验表明，在95%的置信水平下，所提出的方法与对照方法之间没有显著差异。该方法可在30分钟内同时进行96次独立测量，丹磺酰-L-Phe和丹磺酰-D-Phe的检测限（LOD）分别为0.025 μmol L-1和0.075 μmol L-1（3σ）。对丹磺酰-L-Phe（0.78 μmol L-1）进行11次平行测量的相对标准偏差（RSD）为8%。结果表明，基于MIP的CL成像可成为一种用于快速手性识别的有用分析技术。
