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Two peaks emerge from a reversed-phase chromatography column as sketched in the illustration.

According to Equation 23-33, resolution is given by

Resolution=N4(α-1)α(k21+k2)

where Nis plate number, αis relative retention (Equation23-20), and k2 is the retention factor for the more retained component (Equation 23-16).

(a) If you decrease the amount of organic solvent in the mobile phase, you will increase retention. Sketch the chromatogram if retention factors increase but Nand αare constant.

(b) If you change the solvent type or the stationary phase, you will change the relative retention. Sketch the chromatogram ifαincreases but Nandk1are constant.

(c) If you decrease particle size or increase column length, you can increase the plate number. Sketch the chromatogram if Nincreases by (i) decreasing particle size and (ii) increasing column length. Assume αand k2are constant.

Short Answer

Expert verified

a)The sketch of chromatogram if retention factors increase but Nand αare constant:

b) The sketch of chromatogram if αincreases but Nand k1are constant:

c) The sketch of chromatogram if Nincreases by (i) decreasing particle size and (ii) increasing column length.

Step by step solution

01

Definition of retention factor.

The retention factor (Rf) is used in thin-layer chromatography to compare and identify substances. The distance travelled by the compound divided by the distance travelled by the solvent front gives the Rf value of a compound.

02

Step 2: The sketch of chromatogram if retention factors increase but   and  are constant.

a)

When the quantity of organic solvent in the mobile phase is reduced, the retention is automatically enhanced. As a result, ifNandαare held constant, the chromatogram will look like this

03

Step 3: The sketch of chromatogram if αincreases butNandk1are constant.

b)

If the solvent or stationery phase is changed, the relative retention will vary. As a result, the chromatogram will look like this:

04

Step 4: The sketch of chromatogram if N increases by (i) decreasing particle size and (ii) increasing column length.

c)

The chromatogram can be produced as shown below if the particle size is reduced and the plate number is increased.

The chromatogram can be obtained as shown below if the column length is raised and the plate number is increased.

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Most popular questions from this chapter

The chromatogram in Box 25-3 shows the supercritical fluid chromatography separation of seven steroids monitored by three detectors.

(a) In the middle chromatogram, ultraviolet detection provides near universal response for the steroids, whereas in the lower chromatogram the ultraviolet detector provides a selective response for a few of the steroids. How can ultraviolet detection act as either a selective or universal detector?

(b) Why is a sloping baseline observed at 210 nm, but the baseline is flat at 254 nm?

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