Yu Jingjing, Wang Bing, Cai Junlan, Yan Quanping, Wang Sheng, Zhao Ge, Zhao Junwei, Pan Lining, Liu Shaofeng
Zhengzhou Tobacco Research Institute of CNTC Zhengzhou 450001 China
RSC Adv. 2020 Jul 31;10(47):28437-28446. doi: 10.1039/d0ra04096g. eCollection 2020 Jul 27.
Purification and selective enrichment of ultra-low level metabolites in bio-samples is very important for HPLC-MS/MS analysis. A magnetic covalent organic framework ( COFs) (TpPa-1) ( 2,4,6-trihydroxybenzene-1,3,5-tricarbaldehyde--phenylenediamine) material was synthesized and used for a magnetic solid phase extraction (MSPE) method in aromatic amine (AA) metabolites ( 1-naphthylamine, 2-naphthylamine, 3-aminobiphenyl and 4-aminobiphenyl) in urine, and then HPLC-MS/MS was employed for analysis. The morphology, pore structure, surface area, chemical composition, magnetic properties, and thermal stability of the synthesized magnetic COFs were characterized with several analysis techniques, such as nitrogen adsorption-desorption isotherms, HRTEM, FTIR, XRD, and so on. MSPE conditions were optimized and analytical performance of the developed method was characterized. The limit of detection (LOD) and limit of quantification (LOQ) of four AAs ranged from 0.01 to 0.07 ng mL and 0.04 to 0.22 ng mL, respectively. The recoveries of 1-NA, 2-NA, 3-ABP, 4-ABP were in the range of 81.9-105%, 87.8-102%, 101-120%, 88.3-117%, respectively. Good intra-day and inter-day precision were obtained with RSD of less than 5.8% and 9.9%, respectively. Furthermore, the synthesized magnetic COFs absorbent could be recycled in MSPE at least 5 times. Analytical results of AA metabolites in real urine samples with the developed method showed significant difference ( < 0.01) between smokers and nonsmokers. Thus, urinary AA metabolites could be exposure biomarkers for cigarette smoke.
生物样品中超低水平代谢物的纯化和选择性富集对于高效液相色谱-串联质谱（HPLC-MS/MS）分析非常重要。合成了一种磁性共价有机框架（COFs）（TpPa-1）（2,4,6-三羟基苯-1,3,5-三甲醛-对苯二胺）材料，并将其用于尿液中芳香胺（AA）代谢物（1-萘胺、2-萘胺、3-氨基联苯和4-氨基联苯）的磁性固相萃取（MSPE）方法，然后采用HPLC-MS/MS进行分析。用多种分析技术对合成的磁性COFs的形态、孔结构、表面积、化学成分、磁性和热稳定性进行了表征，如氮吸附-脱附等温线、高分辨率透射电子显微镜（HRTEM）、傅里叶变换红外光谱（FTIR）、X射线衍射（XRD）等。对MSPE条件进行了优化，并对所建立方法的分析性能进行了表征。四种AA的检测限（LOD）和定量限（LOQ）分别为0.01至0.07 ng/mL和0.04至0.22 ng/mL。1-NA、2-NA、3-ABP、4-ABP的回收率分别为81.9-105%、87.8-102%、101-120%、88.3-117%。日内和日间精密度良好，相对标准偏差（RSD）分别小于5.8%和9.9%。此外，合成的磁性COFs吸附剂在MSPE中至少可循环使用5次。用所建立的方法对实际尿液样品中AA代谢物的分析结果显示，吸烟者和非吸烟者之间存在显著差异（P<0.01）。因此，尿液中的AA代谢物可能是香烟烟雾的暴露生物标志物。
