| With the rapid development of modern cell biology and molecular biology techniques, new technology is constantly introduced to drug bioactive components detection field, This which also makes it possible to find the active molecules or molecular groups from a large number of natural products, and on the other hand, the application ofmicrofluidic chip analytical techniques, which meet the demand for fast, low cost, high throughput, high intension test requirement, promote the progress of detection method. microfluidic chip analytical techniques has become one of the hot research directions in drug detection of active ingredients. Microfluidic chip analysis have crucial advantagescharacteristics such as with technical analysis miniaturization, good light transmission, non-toxic, micro and nano scale, quantified high pass, integration and good biological compatibility characteristics, which provides a new method and technology platform for the screening of active components from natural products.The topics of this dissertation are presented below:(1)The first chapter mainly introduces the molecular level based microfluidic chip for the new method ofMagnoliae Officinalis extracts antioxidant activity assays.In order to achieve for quantitative testing of Magnoliae Officinalis in the main antioxidant components of magnolol and honokiol, which based on chemiluminescence intensity and antioxidation model, Experimental designed and made a chemiluminescence microfludic chip which integrated four repeating structural unit, At the same time, the antioxidant activity of the extract was also analyzed. The designed micro fluidic chemistry luminescence chip containing serpentine channel, annular efficient mixing channel and ellipse detection zone and structural units. This microfluidic CL chip system can be used to detect multiple samples at a time. In order to verify the feasibility of the chip and the test method, the CL test for magnolol and honokiol standard with chemistry of luminol-H2O2-Cu2+ chemiluminescence system were carried out, and draws a conclusion and get the optimal test conditions when the concentration of Luninol is 0.50 mmol/L, H2O2 concentration is 1.00 mmol/L, Cu2 + concentration is 0.50 mmol/L, velocity is 10.0 ?L/Min. Then the five alcohol extract of Magnolia officinalis were tested. The resultshows that the five extracts have a certain scavenging effect on H2O2, and 1.00 g/mL of five kinds of extracts of antioxidant activity equivalent strength order for: 80% ethanol extract > 60% ethanol extract was more than 40% ethanol extract > water extract> 20% ethanol extract.(2)The second chapter mainly introduces the cell level based microfluidic chip for research on new methods of the natural plant antioxidants of magnolol and honokiol antioxidant activity detection. A novel microfluidic cell culture chip was designed and fabricatedwhich containing 3 x 12 detection window, the chip it can achieve cell continuous culture and cellular antioxidant activity analysis for magnolol and honokiol.The chip mainly comprises a glass substrate, a cell culture layer PDMS and a reagent layer PDMS, a total of 3 parallel cell culture channels, 12 reagent and sample injection channels, 3×12 chemiluminescence detection windows.6 concentrations of 2 samples can be selected at a time on the chip, and the test can be realized on the detector of the fluorescent chemiluminescence micro hole board.After interacting action of 24 h betweenHepG2 cellswith magnolol and honokiol on chip, cell survival rate is more than 90%. With HepG2 cultured on the chipas carrier and luminol aschemical luminescent probe, using 2,2 '- azobis diisobutyl amidine hydrochloride(ABAP) intracellular ROS(reactive oxygen species(ROS), the measured magnolol and honokiol IC50 values were respectively 12.03±0.05?g/mL and 1.54±0.02?g/mL. |
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