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Fabrication and evaluation of magnetic phosphodiesterase-5 linked nanoparticles as adsorbent for magnetic dispersive solid-phase extraction of inhibitors from Chinese herbal medicine prior to ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry analysis.

In the present study, the preparation of the magnetic phosphodiesterase-5 linked Fe3 O4 @ SiO2 nanoparticles was successfully achieved by amide reaction and the magnetic phosphodiesterase-5 linked Fe3 O4 @SiO2 nanoparticles were evaluated as a new adsorbent for magnetic dispersive solid-phase extraction of ligands from medicinal plant samples before the analysis by UHPLC-Q-TOF/MS. The prepared phosphodiesterase-5 linked Fe3 O4 @SiO2 nanoparticles were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, transmission electron microscopy, vibration sample magnetometer and potential laser particle size analyzer. The effects of EDC concentration, incubation time and bead-protein ratio on the amount of immobilized protein were studied. The main experimental parameters affect extraction efficiency of ligands, such as wash times, wash solvents, incubation pH, ion strength and incubation temperature, were investigated and optimized by using echinacoside as a model compound. The absolute recovery of echinacoside was ranged from 98.36%-102.16% in Cistanche tubulosa sample under the optimal extraction conditions. Good linearity was observed in the investigated concentration range of 0.006 mgmL-1 -0.97 mgmL-1 (R2  = 0.9999). The limit of detection was 0.002 mgmL-1 . The RSDs of within-day and between-day precision were less than 2.3%. Due to the excellent magnetic behavior of Fe3 O4 @SiO2 nanoparticles, the proposed method was shown to be simple and rapid. Remarkably, the magnetic phosphodiesterase-5 linked Fe3 O4 @SiO2 nanoparticles could be recycled for ten times with loss of 10% activity.

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