------------------------------------------------------------- 3 ------------------------------------------------------------- 5 -------------- --------------------------------------- 8 -------------------------- 8 --------------------------- 14 -------------------------------- 26 --------------------------------------- 26 --------------------------------------- 26 ---------------------------------------29 ---------------31 --------------------------------------- 31 -------------------------33 1

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(capillary zone electrophoresis, CZE), DNA :, 10-9, 1. 2. 3., 10-9, 4. 3

1 2, 3,,,,,,,, 4

Abstract Almost all fields related to chemistry can not be done without chemical separation. In recent years, with the rapid development of bioengineering, synthetical medicine and environmental science, separation and detection of materials have become the prominent subjects. At the same time, due to the development of life sciences, people pay great attention to health. therefore, it become more and more imminent to make biochemical and medical instruments portable and diminutive. Further more, because the requirements of analysis and separation of complex life system can not be met by the present sensor technique, miniaturization of instruments would bring to significant social and economic benefits Capillary zone electrophoresis has been developed very quickly in recent years. It has unique advantage in analyzing inorganic and biological molecule and has become one of the most important analytical means in medicine, national defence, public security, industry and agriculture. Capillary zone electrophoresis has the following strongpoint: high resolution, high speed, small amount of sample and low cost. Although capillary zone electrophoresis has many advantages, it has drawbacks. First, because capillary zone electrophoresis has a short optical path, the lower limit of its detection is not good. Second, the working voltage is too high to 5

make electrophoretic instruments portable and diminutive. Third. because the diameter of capillary is too small, the injection quantum is as little as 10-9 litre. It is a big problem to inject such little sample into the capillary precisely and automatically. Fourth. the installment of array capillary and inspection of sample is complex in technology. Based on the selective permeance of ionic membrane, focusing electrophoresis makes it possible to focus ion on the membrane, so locale concentration can be achieved. On the basis of different mobility and placement of membrane in different section of electrophoretic channels, separation and concentration of different ions can be realized. In this thesis, a series of apparatus of different types and scales for electrophoresis have been designed and set up and the preliminary separation and concentration experiments have been performed. The following is the main content: 1 Apparatus of focusing electrophoresis have been designed and set up. Using agarose gel and polyacrylamide gel as medium, focusing and separation of protein and organic dyestuff have been conducted and preliminary results have been obtained. Compared with conventional gel electrophoresis, focusing electrophoresis has concentration effect and high resolution 2 Making a good combination of focusing electrophoresis and integrate capillary electrophoresis. That is charactering capillary grooves on the surface of polymethyl, after that, setting up 6

membrane in the grooves, bonding with a cover board. It makes the whole separation process of capillary electrophoresis on a chip. Based on these, we did experiments on focusing and separation of Xylene Cyanol FF and Amido Black 10B, preliminary results have also been obtained. It collects the advantages of focusing electrophoresis and capillary electrophoretic chip. With small amount of sample, automatic detection can be realized quickly and precisely. Because of its low cost of production and manipulation, low pollution, high sensitivity and its portability, it has broad applied foreground in clinic diagnosis, medical appraisal, environmental supervision. 7

19 1809 Re ss (1) 1816 Porret (2) Hardy (3) PH Quincke (4) Helmholtz (5) Smoluchowski 6 V U Picton Linder 7 8 As 2 S 3 1909 Michaelis (9) U PH Abramson Michaelis (10,11) 9

20 20 30 Uppsala Svedberg Jette (12) \Scott (13) Svedberg Tiselius (14) U U U Tiselius U U Tiselius Philpot (15) Svensson (16) Longsworth (17,18) Tiselius 1948 10

50 200 capillary electrophoresis,ce 1942 Martin 16 4 20 Konstantinov 17 Tiselius Everaerts 18 Hjerten 19 1967 UV 70 UV 11

400V/cm 1970 Everaerts 20 1974 Virtenan 21 200 500 m Hjerten 5KV/m, Pretorius 22 1979 Mikkers 23 Everaerts i.d200 m PTEF 16 UV 10 m Everaerts 1981 Jorgenson 24 Lukacs 25 75 m 400000/m, CZE Jorgenson 12

13

(Capillary Electrophoresis) 70 (Micro Opto Electro Mechanical System MOEMS) MOEMS 1992 Manz Harrison [26] MOEMS (1) (2) (3) MOEMS (lab-ona-chip) (4) MOEMS 1 A S(Sample) B(Buffer) W 1 (Waster1) 14

W 2 (Waster2) ( ) W 2 1 2cm S W 1 S W 1 B W 2 1B 2 1992 Manz Harrison [26] 1993 8 SCIENCE [27] [28] [29] [30 31] DNA [32,33] PCR [34,35] [36,37] DNA 1994 Woolley A.T. Mathies R. A. [38] DNA 50 8µm 3.5cm DNA 540s 150 97% 1998 Dieter Schmalzing [39] 15

DNA 200V/cm 14min 400 400V/cm 7min 350 0.5 1999 Shaorong Liu [40] DNA 20min 500 99.4% 1997 Woolley A. T. [41] (Capillary Array Electrophoresis Chip CAE chip) 160s 12 1998 Peter C. Simpson Mathies R.A. [42] 96 96 48 1999 Yining Shi [43] 96 96 10cm ( 1 ) 96 pbr322 2 2000 5 Shaorong Liu [44] 16 DNA 18min 543 99 ( 2 3 ), DNA 10 19 mol 4 5 [45] FITC-Arg FITC-OH 3 16

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