Preparation of restricted access molecularly imprinted polymers based fiber for selective solid-phase microextraction of hesperetin and its metabolites in vivo.

A novel restricted access molecularly imprinted polymers (RAMIPs) fiber was developed for solid-phase microextraction (SPME) of hesperetin and its metabolites in livers of live rats in vivo. Hesperetin as the template, N-isopropylacrylamide as the functional monomer, ethylene glycol dimethyl acrylate as the crosslinker, 2,2-azobisisobutyonnitrile as initiator and bovine serum albumin as the restricted access material were applied in the preparation process. Scanning electron microscopy and Fourier transform infrared spectroscopy were applied to characterize the polymers. The adsorption experiments indicated that RAMIPs-SPME fibers performed high selective recognition property to hesperetin. The selectivity experiment indicated that the adsorption capacity and selectivity of RAMIPs-SPME fibers to hesperetin was higher than that of quercetin, luteolin and baicalein. Macromolecules elimination test showed RAMIPs-SPME fibers could eliminate 94.80%-98.96% of macromolecules, which indicated that RAMIPs-SPME fibers can be used to extract analytes directly from complex biological samples. Furthermore, RAMIPs-SPME sampling combined to ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was applied to capture and identify hesperetin and its metabolites in rat livers in vivo. Finally, hesperetin-7-O-glucuronide, hesperetin-3'-O-glucuronide, eriodictyol and homoeriodictyol were identified as the metabolites of hesperetin. In comparison with the MIPs fibers, commercial PDMS and DVB fibers, RAMIPs-SPME fibers possessed better exclusion effect to macromolecules and higher selectivity to hesperetin and its metabolites. The results demonstrated that the prepared RAMIPs-SPME fiber were proven to be effective tool for the selective adsorption and enrichment of hesperetin and its metabolites from the complex biological fluids.