Skoog/Holler/Crouch Chapter 30

Principles of Instrumental Analysis, 6th ed. Instructor’s Manual

CHAPTER 30
30-1. Electroosmotic flow occurs when a mobile phase in a capillary tube is subjected to a high

potential difference between one end of the tube and the other. For a silica tube, the flow

is generally away from the positive electrode towards the negative. The flow occurs

because of the attraction of positively charge species toward the negative silica surface.

This layer of positive charge is mobile and is attracted toward the negative electrode

carrying with it the mobile phase molecules.

30-2. Electroosmotic flow can be suppressed by reducing the charge on the interior of the

capillary by chemical treatment of the surface.

30-3. In solution, amino acids exist as zwitterions that bear both a positive and a negative

charge. At low pH values, the net effective charge is positive because of the interaction

of hydrogen ions with the amine groups of the amino acid. At high pH values, the net

charge is negative due to the dissociation of the carboxylic acid groups. Thus, at low pH

values, the amino acid molecules will be strongly attracted to the negative electrode while

in basic solution the opposite will be the case.

30-4. Under the influence of an electric field, mobile ions in solution in a capillary are attracted

or repelled by the negative potential of one of the electrodes. The rate of movement

toward or away from the negative electrode is dependent on the net charge on the analyte

and the size and shape of the analyte molecules. These properties vary from analyte to

analyte. Thus, the rate at which molecules migrate under the influence of the electric

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Principles of Instrumental Analysis, 6th ed. Chapter 30

field vary, and the time it takes them to traverse the column varies, making separation

possible.

μ eV (4.31 × 10 −4 )(5000)
30-5. (a) N = = = 1.1×105
2D 2(9.8 × 10 −6 )

(b) N = (4.31×10-4)(10,000)/[2(9.8×10-6) = 2.2×105

(c) N = (4.31×10-4)(30,000)/[2(9.8×10-6) = 6.6×105

30-6. The electrophoretic mobility is given by

μeV 4.31× 10−4 cm 2 s −1 V −1 × 10 × 103 V

v= = = 0.0862 cm s −1
L 50 cm

The electroosmotic flow rate is given as 0.85 mm s-1 = 0.085 cm s-1

Thus, the total flow rate = 0.0862 + 0.085 = 0.1712 cm s-1, and

t = [(40.0 cm) / 0.1712 cm s-1)] × (1 min / 60 s) = 3.9 min

30-7. In micellar electrokinetic capillary chromatography surfactants are added to the

operating buffer in amounts that exceed the critical micelle concentration. The

components of a sample added to this system distribute themselves between the aqueous

phase and the hydrocarbon phase in the interior of the micelles according to their

distribution coefficients, similar to partition chromatography. The negatively charged

micelles do exhibit electrophoretic flow but at a much slower rate than the buffer. Thus,

MECC is a true chromatography, where CZE separates sample components simply on the

basis of ion mobility in a homogeneous buffer solution.

30-8. The major advantages of MECC include higher column efficiencies and the ease with

which pseudostationary phase can be altered.

30-9. The order of elution should be B+ followed by A2+ followed by C3+.

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Principles of Instrumental Analysis, 6th ed. Chapter 30

30-10. The elution order in sedimentation FFF is determined by particle size and mass.

30-11. FFF methods are particularly well suited to high molecular weight materials such as

polymers, large particles and colloids. In addition, no packing material or stationary

phase is needed for separation to occur avoiding undesirable interactions between the

packing material and the sample constituents. Finally, the geometry and flow profiles in

FFF are well characterized allowing fairly exact theoretical predictions of retention and

plate height to be made.