UltiMate3000DGLC双三元液相色谱的在线柱后衍生技术_

selectivity could be achieved by this method according to the selective complementarity of the two columns．In the experiment ，ACN-H 2O was used as the SPE mobile phase with a flow rate of 1mL /min and ACN-10mmol /L NH 4Ac was used as the analytical mobile phase with a flow rate of 1mL /min．Serum samples were injected directly into the SPE column and the biological matrix was washed out with the loading solvent．By rotation of the switching valve ，Clozapine ，Quetiapine and Ｒisperidone were eluted from the SPE cartridge in the back-flush mode and transferred to the analytical column by the chromatographic mobile phase．The whole time of the online SPE purification and chromatographic separation of the analytes was 18min．Calibration curve of Clozapine with good linearity （r =0．9996）was obtained in the range of 10－1800μg /L in human serum ，and the recoveries at low ，medium and high concentration levels were 118．4%，105．0%and 105．4%．Calibration curve of Quetiapine with good linearity （r =0．9997）was obtained in the range of 3．6－640μg /L in human serum ，and the recoveries at low ，medium and high concentration levels were 112．8%，101．1%and 101．5%．Calibration curve of Ｒisperidone with good linearity （r =0．9995）was obtained in the range of 0．71－128μg /L in human serum ，and the recoveries at low ，medium and high concentration levels were 100．7%，97．2%and 98．8%．In conclusion ，the established automated online SPE-HPLC-UV method demonstrated good performance in terms of linearity ，specificity ，limits of quantification ，and was successfully utilized to quantify Clozapine ，Quetiapine and Ｒisperidone in human serum．

Keywords High performance liquid chromatography ；Online solid phase extraction ；Clozapine ；Quetiapine ；

Ｒisperidone ；Serum

（Ｒeceived 18July 2014；accepted 14October 2014）櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫櫫

UltiMate 3000DGLC 双三元液相色谱的在线柱后衍生技术

衍生化是指被测物质与相应的试剂发生化学反应，改变被测物质的化学和物理性质，提高被测物质的检测灵敏度，改善被测物质混合物的分离度，从而达到利于分析的目的。衍生化在HPLC 分析中应用广泛，其中在线柱后衍生使用居多。传统的在线柱后衍生需要一个独立的泵来输送衍生试剂以实现衍生的目的，但这些额外的独立装置有时由于与液相色谱系统本身不能进行很好的联用，往往导致运行不甚理想。赛默飞UltiMate 3000DGLC 双三元液相色谱由于具有两个独立的泵和流路，一个泵进行分析，另外一个泵可以完成如输送衍生试剂等的辅助功能，可以很好地实现在线柱后衍生应用，而且这些功能很多情况只需要一个简单的柱后三通或反应管的连接就可以实现。此外，在HPLC 和MS 联用时，为了增强离子化效率，需要另外的流路泵入甲酸或乙腈，使用UltiMate 3000DGLC 双三元液相色谱也可以简单方便地实现这样的功能。例如食品中黄曲霉毒素的检测，使用UltiMate 3000DGLC 双三元液相色谱的右泵作为分析泵，左泵做衍生泵，以0.05%碘溶液作为衍生反应试剂，一套系统即可方便自动化地实现在线柱后衍生，提高黄曲霉毒素的检测灵敏度，以满足法规的检测要求。7281第12期沈广虎等：在线固相萃取-高效液相色谱法同时测定人血清中氯氮平、奎硫平和利培酮的含量