Research Of High-performance Hydrogen Peroxide Electrochemical Sensor Used In Evaluation Of Anti-cancer Activity Of Paclitaxel

Small molecule reactive oxygen species H₂O₂ participates in many physiological processes in living organisms.It often exists in biological processes as a ubiquitous intracellular messenger or receptor signal in various cells,affecting multiple signal transduction pathways and cell growth regulation.detection index.Taking the electrochemical detection of small molecules as a clue,the traditional Chinese medicine monomer paclitaxel as the experimental object,and the different concentrations of H₂O₂ released by liver cancer cells with different survival rates after stimulation with different concentrations of paclitaxel solution as the evaluation index to evaluate the anticancer efficacy of paclitaxel.The detecting results of electrochemical sensor were the same as the results of conventional detection method CCK-8 method and fluorescence detection method.The same results determined that the new electrochemical sensor is a feasible route to evaluate the efficacy of traditional Chinese medicine.In order to improve the detection sensitivity of electrochemical sensors,functionally modified metal organic frameworks（MOFs）nanomaterials are used as sensor working electrode modification materials.The excellent properties such as surface area,morphology and size can be adjusted to build a working electrode with high capture ability,which improves the sensitivity and specificity of the sensor.In order to screen out high-sensitivity modified materials,this study screened metal-organic frameworks（MOFs）with two-dimensional planar（2D）and three-dimensional（3D）morphologies by linking the same metal ions through the different morphologies of nanomaterials.),as well as the catalytic performance of MOFs linked with different metal ions（Co,Cu,Zn）of the same two-dimensional planar（2D）morphology for H₂O₂,and their application as supporting materials in the construction of H₂O₂electrochemical detection sensors.A non-enzymatic electrochemical sensor for H₂O₂ based on MOFs and its derivatives was constructed to realize the real-time detection of H₂O₂ released by cancer cells under drug stimulation.The rapid evaluation of the efficacy of traditional Chinese medicine is the key of the new traditional Chinese medicines screening.Conventional drug efficacy evaluation methods cannot evaluate the effects of drugs on organisms in real-time,and the evaluation process is time-consuming,involving many reagents and instruments,which consumes human costs and material resources.In order to meet the need of rapid evaluation of the efficacy of traditional Chinese medicine,this work focuses on electrochemical sensing technology.In this experiment,the real-time detection of small molecule compounds released by cells is used as the evaluation index.The cells stimulated by the monomer of traditional Chinese medicine,and a new electrochemical sensor is constructed to evaluate the efficacy of traditional Chinese medicine.Rapid detection by electrochemical sensors achieved the same results as conventional drug efficacy evaluation,the same results proved that the rapid evaluation of traditional Chinese medicine efficacy with the help of new electrochemical sensors is a feasible solution.The main research contents of this topic include the following two aspects:（1）Highly sensitive electrochemical sensors constructed by cobalt-based metal-organic frameworks for detecting cell-released small moleculesIn order to evaluate the efficacy of traditional Chinese medicines through electrochemical sensors,to enable the electrochemical sensors to detect low-concentration small molecule compounds released by cells in real time,screening high catalytic materials is an effective method to improve the sensitivity of the sensor,the electrochemical properties of noble metals were investigated.The comparison showed that when the ratio of Au:Pt=3:7（Au₃Pt₇NPs）,the catalytic activity of noble metal nanoparticles（NPs）for H₂O₂ was the best,and then compared with cobalt metal.Two new metal-organic framework materials with different morphologies formed by ion linking:classical three-dimensional cobalt-based metal-organic framework 3D Co-MOF（ZIF-67）and classical two-dimensional cobalt-based metal-organic framework 2D Co-MOF（Co-TCPP）.Comparing the electrochemical properties of the two and their electrocatalytic effects on H₂O₂,Co-TCPP has the largest cyclic voltammetry peak current value and current-time（I-t）curve response to H₂O₂,which indicating that it has the best catalytic effect.The Au₃Pt₇ NPs was electrodeposited onto 2D and 3D Co-MOFs,respectively.Under the optimal conditions,the signal and concentration of Au₃Pt₇/Co-TCPP/GCE for H₂O₂concentration detection ranged from 0.1μM to 60 m M.Good linearity was observed with a lowest detection limit（LOD,S/N=3）of 0.020μM.The electrode constructed by this method has strong anti-interference,good repeatability and reproducibility,and has excellent stability.It has successfully realized the detection of H₂O₂ released by cervical cancer cells Hela cells,and initially achieved real-time detection of drug stimulation by cancer cells.（2）A new electrochemical sensor was constructed to evaluate the anticancer effect of the traditional Chinese medicine paclitaxel.Comparative screening of metal-organic frameworks M-TCPP with the same morphology and different metal ion linkage points.M represents three different metal elements Co,Cu,Zn.M-TCPP was used to support the noble metal nanoparticles Au₃Pt₇ NPs（Au:Pt=3:7）to construct the new H₂O₂ enzyme-free electrochemical sensor.Using the I-t curve method,it was found that Cu-TCPP had the best catalytic effect on hydrogen peroxide and the strongest reducing signal appealed.The electrochemical testing results were similar as the TMB detecting result.The resulting two-dimensional Cu-MOF is used as a support material,and Au₃Pt₇ is stably combined on the surface of the Cu-TCPP/GCE electrode by electrodeposition.As the two materials collaborate worked,the sensor showed good selectivity and sensitivity.After we find optimal conditions,the sensor can detect H₂O₂ in a linear range of 10 n M-80m M with a detection limit of 3.1 n M（S/N=3）.The electrochemical sensor was used to detect H₂O₂ which released from Hep G2 cells after the Hep G2 cells stimulated by different concentrations of paclitaxel.Tells from the results,different concentrations of paclitaxel made the different rates of Hep G2 cells.By AA stimulating,H₂O₂ released from Hep G2 cells could be detected by the constructed Au₃Pt₇/Cu-TCPP/GCE working electrode.Comparing with conventional methods,the results are consistent.It shows that the new electrochemical sensor constructed in this work can effectively evaluate the anticancer effect of the traditional Chinese medicine monomer paclitaxel,and provides a new idea for the rapid evaluation of the efficacy of traditional Chinese medicine.