吴翊民, 吴庆政, 王晓春, 谢增鸿, 吴晓苹

(福州大学化学化工学院,福建福州350108)

亲水作用加压毛细管电色谱-激光诱导荧光检测法分析痕量核黄素类物质

吴翊民, 吴庆政, 王晓春, 谢增鸿, 吴晓苹＊

(福州大学化学化工学院,福建福州350108)

一种新型的亲水作用毛细管电色谱(H I-CEC)整体柱被应用于加压毛细管电色谱-激光诱导荧光检测(pCECL IF)联用法对核黄素类物质的分离分析。采用自组装的pCEC-L IF系统,实现了对痕量核黄素(RF)、黄素单核苷酸(FMN)和黄素腺嘌呤二核苷酸(FAD)的快速分析。在最优的分离检测条件下,3种化合物在8.0m in内完全分离,RF、FMN和FAD的检出限(LOD,S/N=3)分别为5.0×10-11mol/L、8.0×10-10mol/L和2.5×10-9mol/L,测定线性范围可达3个数量级,精密度良好。方法简便、全分析时间短、灵敏度和选择性高,血清样品分析实验结果良好,可望进一步应用于体液及细胞中核黄素类物质的痕量检测。

加压毛细管电色谱;亲水作用;整体柱;激光诱导荧光检测;核黄素类物质;血清

Abstract:A novel hydrophilic interaction capillary electro Chromatographic(H I-CEC)monolith with covalently bonded zwitterionic functional groups was applied for the separation of riboflavins(RF)and its derivatives.with a home made pressurized capillary electro Chromatographylaser induced fluorescence detection(pCEC-L IF)system,trace levels of RF and its derivatives,flavin mononucleotide(FMN)and flavin adenine dinucleotide(FAD),can be baseline separated within8.0m in inisocratic elution m ode.The effect of experimental parameters on separation was investigated.Under the optimum conditions,analytes could be determined over nearly three orders of magnitudes with the detection limits(LOD s)as lowas5.0×10-11mol/L(RF),8.0×10-10mol/L(FMN),2.5×10-9mol/L(FAD),and the relative standard deviations(RSD s)were less than8.2%.This method is rap id,simple,repeatable and more sensitive than the most of reported methods,and satisfied results has been achieved in serum sample.Furthermore,it can be further applied for trace analysis of RF and its derivatives in biological fluid and cells.

Key words:pressurized capillary electro Chromatography(pCEC);hydrophilic interaction(HI);monolith;laser induced fluorescence detection(LIF);riboflavin and its derivatives;serum

毛细管电色谱(CEC)具有高效液相色谱(HPLC)和毛细管电泳(CE)的综合优势[1,2],近年来随着检测方法和整体柱制备技术的迅速发展[3-5],CEC及联用分析技术及其在生命科学、环境等领域[6-10]的应用已成为目前色谱领域的研究热点之一。反相(RP)分离模式是CEC所采用的主要模式,然而在分离极性物质时,由于RP固定相表面不易与流动相充分润湿,容易产生一系列的问题,如在CEC分离过程中产生气泡、中断分离甚至造成固定相塌陷等[11]。加压毛细管电色谱(pCEC)很好地解决了气泡问题。随着填充柱技术研究的深入,逐渐建立了一些更适合极性物质分离的CEC模式,如亲水作用CEC(HI-CEC)[12,13],已应用于多肽、蛋白质、核苷、核苷酸和寡糖等生化样品的分离分析。……