新型固相吸附剂-固相微萃取-液相色谱-质谱联用检测血液和尿液中的喹诺酮类抗生素

Quinolones antibiotics are widely used to treat bacterial infections in clinical care. But the inappropriate use of these drugs, which are excreted either as parent compounds or as metabolites via urine and faeces, can result in the environmental pollution and threating human health. Therefore, rapidly, easily and accurately analytical methods are ugently requried. This project focused on synthetising new solid absorbents with perfect selectivity, developing new solid phase microextraction (SPME) method to extracting and concentrating quinolones antibiotics and their metabolites in blood and urine samples, exploring the efficiency of various SPME methods on extraction and enrichment, selecting the optimal absorbents and micrioextraction method and determing the quinolones antibiotics and their metabolites after extraction by liquid chromatography-mass spectrometry. The project aims to make the effective and trace analyses of quinolones antibiotics and their metabolites in biological sample, and further provide methodological basis for point-of-care testing and theoretical basis for clinical medication. SPME methods can efficiently reduce agents and time consumption, moreover, eliminate matrix interference when extracting the target analytes. Therefore, SPME methods are very suitable for complex biological sample preparation. The successful study of this project will bring in vital theoretical and application value for standard method for biological sample preparation, determination method and clinical laboratory of quinolones antibiotics.

喹诺酮类抗生素在临床上广泛用于抗菌治疗。但是此类抗生素的不合理使用使大量的药物和代谢产物排泄到环境中，对环境造成污染并危害人体健康。因此，急需此类抗生素的快速、简单、准确的分析方法。本项目拟自合成高选择性的固相吸附剂,建立新型的固相微萃取方法萃取富集血液和尿液中的喹诺酮类抗生素及其代谢产物，探索各种固相微萃取方法对萃取富集效率的影响，选取最佳的吸附填料及微萃取方法与液相色谱-质谱联用对富集后的目标分析物进行特异灵敏测定，实现临床生物样品中喹诺酮类抗生素及其代谢产物的高效、痕量分析，为此类抗生素的床边监测提供方法学依据，也为临床用药提供一定的理论基础。固相微萃取方法能有效地减少试剂的使用、缩短萃取时间，而且在萃取目标分析物的同时消除基质干扰，特别适用于复杂生物样品前处理。本研究的顺利实施将对生物样品中喹诺酮类抗生素的萃取富集标准方法、测定方法及临床检验等都具有重要的理论意义和应用价值。

由于对方法可靠性、灵敏度、分析速度和样品通量方面的要求非常苛刻，环境和生物样品分析方法的开发在过去几年中变得越来越具有挑战性。本项目重点研究了各种液相微萃取方法，包括盐析辅助液液萃取（Salting-out assisted liquid-liquid extraction, SALLE）、分散液液微萃取（Dispersive liquid-liquid microextraction, DLLME）、浊点萃取（Cloud point extraction, CPE）及微萃取方法联用来萃取和富集多介质环境样品中的污染物（如空气、水、土壤、人尿和血液），并通过液相色谱—质谱（Liquid chromatography-mass spectrometry, LC-MS）和气相色谱—质谱（Gas chromatography-mass spectrometry, GC-MS）测定污染物。该项目综述了各种生物样品的样品前处理方法。同时，该项目成功建立了SALLE-DLLME-GC-MS、DLLME- LC、SALLE-GC-MS和CPE-LC方法用于测定尿液、血液、空气、土壤和环境水样中的拟除虫菊酯类杀虫剂、喹诺酮类抗生素和磺胺类抗菌药物。在最佳萃取及测定条件下，方法具有低的检测限（ng/mL）、高的回收率（70%-120%）和宽的线性范围（2-3个数量级），而且方法简单、快速、灵敏。项目结果表明微萃取-色谱联用方法可以成功用于多介质环境样品中各种环境污染物的测定。该项目将为环境污染物的生物和环境样品前处理方法、测定方法和临床实验室的标准方法带来重要的理论和应用价值。