Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation
description
Transcript of Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation
![Page 1: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/1.jpg)
Ch. 30
Capillary Electrophoresis, Capillary Electrochromatography
and Field - Flow Fractionation
Electrophoresis = ?
Capillary Electrophoresis = ?
![Page 2: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/2.jpg)
電 泳
1948 Nobel Prize
Q / d : charge-to-size ratio
Swedish chemist Serum proteins
Electrophoresis: a separation method based on
the differential rate of migration of charged species in a
buffer solution across which has been applied a dc
electric field.
![Page 3: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/3.jpg)
Slab electrophoresisBio macromolecules:
Proteins (enzyme, hormones, antibodies)
Nucleic acids (DNA, RNA)
L. Stryer, Biochemistry, 4th ed., W. H. Freeman, 1999, p. 47.
平板
![Page 4: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/4.jpg)
F. Rouessac and A. Rouessac, Chemical Analysis, Wiley, 2000, p.112
![Page 5: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/5.jpg)
毛細管電泳
Human Genome: sequence for human DNA
![Page 6: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/6.jpg)
![Page 7: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/7.jpg)
Systematic apparatus for capillary electrophoresis
15 ~ 30 kV
25 ~ 75 μm i.d.
~ 50 cm long
G. D. Christian, Analytical Chemistry, 6th ed., John Wiley, 2004, p. 632
vs. slab eletrophoresis:no joul heating effect
![Page 8: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/8.jpg)
FIGURE 30-1 Schematic of a capillary electrophoresis system.
![Page 9: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/9.jpg)
D. C. Harris, Quantitative Chemical Analysis, 6th ed., 2003, p. 655.
![Page 10: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/10.jpg)
FIGURE 30-2 Charge distribution at a silica-capillary interface and resulting electroosmotic flow.
buffer solution
e.g. Na+Ac
-
Acts as a pump
帶動整體溶液電 滲 流
OH pH > 3 O-
Si Si
electricaldouble layer
Microfluid
微流體
![Page 11: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/11.jpg)
D. C. Harris, Quantitative Chemical Analysis, 6th ed., 2003, p. 658.
+ > -
+ = -
![Page 12: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/12.jpg)
FIGURE 30-4 Velocities in the presence of electroosmotic flow. The length of the arrow next to an ion indicates the magnitude of its velocity; the direction of the arrow indicates the direction of motion. The negative electrode is to the right and the positive electrode to the left of this section of solution.
In general,
νeo > νe
Separation is based on differences in electrophoretic mobility.
電滲流
電泳 ++ -
charge / size
migration velocity
淨移動速率
![Page 13: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/13.jpg)
![Page 14: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/14.jpg)
![Page 15: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/15.jpg)
FIGURE 30-3 Flow profiles for liquids under (a) electroosmotic flow and (b) pressure-induced flow.
electroosmotic hydrodynamic
parabolicflat
pump
+ -
![Page 16: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/16.jpg)
D. C. Harris, Quantitative Chemical Analysis, 6th ed., 2003, p. 658.
pump
![Page 17: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/17.jpg)
ms
D. C. Harris, Quantitative Chemical Analysis, 7th ed., 2007.
ms
![Page 18: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/18.jpg)
D. C. Harris, Quantitative Chemical Analysis, 6th ed., 2003, p. 656.
![Page 19: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/19.jpg)
van Deemter equation:
D. C. Harris, Quantitative Chemical Analysis, 6th ed., 2003, p. 656.
![Page 20: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/20.jpg)
Advantages of CZE:
• High separation efficiency (N > 105)
• Low reagent consumption
• Low sample consumption (< 10 nL)
![Page 21: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/21.jpg)
Sample introduction in CZE:
1. Electrokinetic injection
2. Pressure injection
a. pressure
b. vacuum
c. height difference (gravity)
![Page 22: Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation](https://reader036.fdocuments.in/reader036/viewer/2022081506/568157f3550346895dc5738b/html5/thumbnails/22.jpg)
TABLE 30-1 Detectors for CE
(UV/Vis)