Chemiluminescence Paper-based Microfluidic Research

Microfluidic chip integrates the selection, preparation, reaction, separation, detection and other basic operation of biological sample in chemical and biological fields into a several square centimeters (or even smaller) of microchip. It controls the flow by micro-channel network in order to complete the different chemical or biological reaction process, and analyzes the reaction products. It provides a new platform for the creation of new situation of biochemical analysis. It is based on analytical chemistry, microfabrication, micro fluid drive or control and detection technology. It makes micro-channel as structure feature, and makes life science as the main application object. The paper-based microfluidic is introduced by Martinez firstly and develops quickly in recent years. The paper-based microfluidic channels patterned by photolithography, inkjet printing or wax printing, and it has been successfully applied in the fields of colorimetric analysis, electrochemical analysis and biological sample analysis. Because it is small, light-weight, portable, single-use, disposable, low cost, and it requires small volumes of sample, and has quick analysis speed, and it can measure multiple samples in parallel, it has been increasingly applied in chemical and biological fields, and has very good development prospect. Herein we describe a simple method for patterning paper-based microfluidic with filter paper, and combine it with chemical luminescence. Accordingly we study the chemiluminescence paper-based microfluidic initially.Three chapters are included in this thesis. The first chapter is review, which mainly introduce the history, poduction methods, detemination technology and application of microfluidic chip, and especially, the production methods and application of paper-based microfluidic are introduced. The second chapter and third chapter are research reports, and the main contents are as follows:1. A simple method for patterning paper-based microfluidic was described. We discussed some problems occurred-in the fabrication process of paper-based microfluidic by luminol-potassium ferricyanide-isoniazid chemiluminescence reaction. Because one of the most important problem in the study of chemiluminescence is repeatability, we did some experiments about paper-based microfluidic applied in chemiluminescence detection,and we also investigated some factors effecting repeatability. Finally we detected isoniazid with better result, and it is further validated the feasibility of the paper-based microfluidic applied in chemiluminescence detection.2. In pH=8.0 of KH2PO4-Na2HPO4-12H2O buffer solution, copper (Ⅱ) can coordinate with calcein, and cause the fluorescence intensity of calcein solution decrease. Because the affinity between copper (Ⅱ) and acetylcysteine was very strong, it can seize copper ions from calcein-copper (Ⅱ) complex and make calcein free, thus make the fluorescence recover, and the recovery degree of fluorescence was linear related to the concentration of acetylcysteine. Accordingly, we establish a new method of measuring acetylcysteine in the range of 6.0×10-6-1.4×10-5 mol/L. The detection limit was 4.0×10-6 mol/L. The relative standard deviation was 0.91% for the 11 replicate determinations of 1.0×10-5 mol/L acetylcysteine solution. The method has been sucessfully applied to the determination of acetylcysteine in pharmaceutical preparations with satisfactory result.