Analysis of biologically active substances new method of separation

Title: Analysis of biologically active substances new method of separation Author: Zhang Xiaoqing Degree-granting units: Chongqing Medical University Keywords: tryptophan;; kynurenine;; indole 2,3 - dioxygenase;; high-performance liquid chromatography;; Kawasaki;; immunoassay;; beads;; electrochemical
Science;; tumor markers;; microfluidic Abstract: Separation and analysis of bioactive substances in the modern scientific analysis has been showing an increasingly important role, and is Magnetic lifter widely used in food safety
All, environmental monitoring, clinical testing, drug analysis, medicine and health, and quality control and other areas. In these areas often need to
Complex system of one or more components of the separation analysis, such as traditional Chinese medicine analysis of some of the active ingredient concentration determination, clinical diagnosis
In the disease-related markers, food analysis for the detection of melamine in milk and so on. This allows simple, rapid, sensitive
, Low-cost method of separation and analysis is particularly important. The study on the establishment of the separation of bioactive substances to carry out the following analysis
Work: 1 high-performance liquid chromatography - application design wavelength UV detection for simultaneous determination of plasma tryptophan and kynurenine
And its application In this study, high-performance liquid chromatography - application to set wavelength UV detector set up plasma tryptophan and kynurenine
The simultaneous determination method is simple, rapid, sensitive, specific and so on. In the Agilent Hypersil ODS (125mm ×
4.0mm, 5μm) column can be completed within 6 minutes of plasma tryptophan and kynurenine, while separation and determination of two components. Outflow of fluid through the purple
Outside the detector wavelength http://www.999magnet.com/products/131-magnetic-lifter program designed to detect, 0-4 minutes to detect the wavelength set to 360nm kynurenine, 4-6 minutes is set to 278nm wavelength
To detect tryptophan, so that these two components can be realized in a flow process in the same test. The method of determination of linear kynurenine
Range is 0.269 to 21.20μmol / L, the linear range of tryptophan is 3.35 to 678.0μmol / L. Kynurenine and tryptophan detection
Limits are 0.028μmol / L and 0.053μmol / L. Precision and recovery requirements are in line with clinical examination. This method is used to analyze
Kawasaki disease, Henoch-Schonlein purpura, nephrotic syndrome, acute nephritis in children of patients with plasma tryptophan and kynurenine concentrations. The results showed Chuan
Kawasaki disease group and control group, the indicators are significant differences between the other group of diseases and the control group, no significant difference. 2. Functionalized superparamagnetic
Of beads used for tumor markers The use of superparamagnetic beads, combined with screen-printed electrode (SPCE) for the working electrode
Electrochemical detection techniques, proposed a new use of magnetic sensors fixed biological components of the immune, tumor marker CA199 for the sub-
From detection. The sensor is by the end of the epoxy-modified superparamagnetic nano-beads on the surface binding chain avidin, the use of chain enzyme
Avidin and biotin reaction, the biotin-labeled antibodies attached to the bead surface, then add by off-line incubation of the samples and horseradish peroxide
Enzyme-labeled antibody material (HRP-anti-CA199), so that HRP was successfully connected to the surface of superparamagnetic beads, to be fixed by the magnetic field
Set at the SPCE surface, adding o-phenylenediamine. Hydrogen peroxide system for electrochemical detection. Electrochemical response signal and the concentration of CA199 antigen
Proportional to the linear range of 5.0 ~ 120U/mL. This method does not require immobilization of reagents, is simple, sensitive, low cost.
3. Microfluidic chip electrochemical detection in the immune Analysis With the analytical chemistry, biology, electronics, computer technology
Surgery and the rapid development of micro-processing technology, new analytical instruments and methods continue to be used in laboratory medicine, laboratory medicine has to be accordingly
Rapid development. Laboratory medicine has been gradually raised to the immediate test (point-of-caretesting, POCT), home testing or field inspection
Measurement direction, in order to achieve fast, easy and specimen testing. Recently developed microfluidic immunoassay miniaturization of the instrument and
Portable and has a good prospect, so it possible to achieve the goal of POCT. Microfluidic immunoassay micro-processing technology by
Build on the chip material micron channels for immunoassay. Microfluidic immunoassay because of its unique nature, can greatly improve the often
Regulations immunoassay performance: (1) The micro-channel structure larger than the surface area, can greatly reduce the immune response and improve analysis efficiency. Microfluidic
Control analysis of multi-chip within a few seconds or tens of seconds to complete the separation and detection components. (2) The channel size is small, significant savings can be
Precious immune reagents consumption, reduce testing costs. Microfluidic analysis of the sample volume has been reduced to the nanoliter or even lower levels. (3)
Portable instrument is easy to realize miniaturization and automation, is expected for POCT. In this study, a microfluidic channel in the immune response,
Printed electrodes integrated on the chip will be implemented on microfluidic chip immunoassay device for the integration of electrochemical detection assay of tumor markers
. Study microfluidic chip power injection technology, immunoassay and microfluidic channels combined end-column electrochemical detection, trying to build a
Sample collection, separation, detection in one simple and easy portable microfluidic immunoassay chip, serum tumor markers. Degree Year: 2009