柱前稀释固相萃取-超临界流体色谱质谱在线联用技术及其在手性药物分析中的应用

The study of coupling technique of sample pretreatment and chromatography has become the frontier in the current analysis science. Supercritical fluid chromatography (SFC) is a new type of chromatography which has been developed rapidly in recent years. SFC has many advantages such as low cost, green, fast and efficient, and especially in chiral analysis. However, SFC also meets some problems such as difficulty to analysis the aqueous samples directly and susceptibility to matrix interference. In this project, a pre-column dilution on-line coupling technique based upon solid-phase extraction and supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS) is proposed to be established, and will be applied to the fast, highly sensitive and selective online analysis of chiral drugs. The realization of this online technique would not only solve the problem of analysis the aqueous samples directly, but also decrease the matrix interference and improve the sensitivity and the automation. This online system is realized by adopting the pre-column dilution and valve switching technique from our previous research, which increases the injection volume and realizes the pressure separation of SPE and the SFC systems. The features of the online system and the online analytical method would be systematically investigated and compared with the offline technique. Furthermore, the features of the pre-column dilution and the mechanism would also be investigated. Finally, the online technique would be applied to the analysis of chiral drugs in biological samples. It is strongly believed that this research is of great importance for SFC in the rapid, high sensitive analysis of chiral drugs and the automation of the chiral analysis.

样品前处理与色谱分析联用的研究已成为当前分析科学领域的前沿课题。超临界流体色谱是近年来快速发展的一种新型色谱技术，具有经济、环保、快速、高效等优点，尤其在手性分析方面优势明显，但是也存在难以直接分析水相样品、易受基质干扰等问题。本项目拟发展一种基于柱前稀释的固相萃取-超临界流体色谱质谱在线联用技术，并应用于手性药物的快速、高灵敏、高选择性在线分析。该联用技术不仅可实现水相样品直接进样，而且显著降低基质干扰、提高分析灵敏度和自动化程度。该联用技术采用课题组前期研究的柱前稀释柱头聚焦技术和阀切换技术显著增大进样溶液体积、实现固相萃取与色谱系统的压力分离，从而实现在线联用，并对联用系统和联用方法进行系统考察，研究柱前稀释的特点和机理，并与离线技术进行对比，最后应用于生物样品中手性药物的分离分析。本研究对推动超临界流体色谱在复杂样品中手性药物的高灵敏度快速检测和自动化分析具有十分重要的意义。

样品前处理与色谱分析联用的研究已成为当前分析科学领域的前沿课题。超临界流体色谱是近年来快速发展的一种新型色谱技术，具有经济、环保、快速、高效等优点，但是也存在难以直接分析水相样品、易受基质干扰等问题。本项目建立了超临界选择性萃取-超临界流体色谱质谱在线联用系统，成功应用于血浆中22种手性药物分析：引入了变相聚焦和选择性萃取步骤，克服了在线超临界流体萃取-超临界流体色谱中峰展宽和基质干扰的问题。该方法标准曲线线性相关系数大于0.99、LOD达0.0200 -43.6 μg/L、方法准确度为79.0-114.5%、日内和日间精密度RSDs%分别小于5.8%和13.9%；建立了固相脱水萃取-超临界流体色谱质谱在线联用系统，成功应用于人尿中45种兴奋剂检测：引入了全自动脱水步骤以及解吸液柱前稀释步骤，克服了超临界流体色谱不能直接分析含水样品的缺点。该方法标准曲线线性相关系数大于0.99、LOD达0.0380 ng/L-1.24 μg/L、方法准确度为80.8-119.7%、日内和日间精密度RSDs%分别小于10.8%和15.4%。与其他方法相比，能有效减弱基质效应、提高灵敏度和准确性；建立了冻干超临界流体萃取-超临界流体色谱质谱在线联用系统，成功应用于香草兰豆荚中香草醛及其类似物以及乳腺癌细胞中7种内源性脂质和抑郁症大鼠脑组织中的23种炎症相关脂质的分析。方法学验证结果均符合要求，且与其他方法相比，在线方法的萃取效率更高、分析速度更快、灵敏度和准确度更高。本项目发展的几种联用技术不仅可实现高含水样品的直接萃取分析，而且显著降低基质干扰、提高分析灵敏度和自动化程度，对推动超临界流体色谱在复杂样品中的高灵敏度快速检测和自动化分析具有十分重要的意义。