Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography

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Presentation transcript:

Ch. 30 Capillary Electrophoresis, Capillary Electrochromatography and Field - Flow Fractionation Electrophoresis = ? Capillary Electrophoresis = ?

電泳 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. Q / d : charge-to-size ratio 1948 Nobel Prize Swedish chemist Serum proteins

平板 Slab electrophoresis Bio macromolecules: Proteins (enzyme, hormones, antibodies) Nucleic acids (DNA, RNA) 平板 L. Stryer, Biochemistry, 4th ed., W. H. Freeman, 1999, p. 47.

F. Rouessac and A. Rouessac, Chemical Analysis, Wiley, 2000, p.112

毛細管電泳 Human Genome: sequence for human DNA

Systematic apparatus for capillary electrophoresis 25 ~ 75 μm i.d. ~ 50 cm long 15 ~ 30 kV vs. slab eletrophoresis: no joul heating effect G. D. Christian, Analytical Chemistry, 6th ed., John Wiley, 2004, p. 632

FIGURE 30-1 Schematic of a capillary electrophoresis system.

D. C. Harris, Quantitative Chemical Analysis, 6th ed., 2003, p. 655.

Acts as a pump 帶動整體溶液 buffer solution e.g. Na+Ac- Microfluid 微流體電滲流 electrical double layer OH pH > 3 O- Si Si FIGURE 30-2 Charge distribution at a silica-capillary interface and resulting electroosmotic flow.

+ = - + > - D. C. Harris, Quantitative Chemical Analysis, 6th ed., 2003, p. 658.

- + + νeo > νe In general, 電滲流電泳 Separation is based on differences in electrophoretic mobility. charge / size migration velocity 淨移動速率 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.

parabolic flat electroosmotic hydrodynamic + - pump FIGURE 30-3 Flow profiles for liquids under (a) electroosmotic flow and (b) pressure-induced flow.

pump D. C. Harris, Quantitative Chemical Analysis, 6th ed., 2003, p. 658.

ms ms D. C. Harris, Quantitative Chemical Analysis, 7th ed., 2007.

D. C. Harris, Quantitative Chemical Analysis, 6th ed., 2003, p. 656.

van Deemter equation: D. C. Harris, Quantitative Chemical Analysis, 6th ed., 2003, p. 656.

Advantages of CZE: High separation efficiency (N > 105) Low reagent consumption Low sample consumption (< 10 nL)

Sample introduction in CZE: Electrokinetic injection Pressure injection a. pressure b. vacuum c. height difference (gravity)

TABLE 30-1 Detectors for CE (UV/Vis)