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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(-1)(k21+k2)

where Nis plate number, is 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 are constant.

(b) If you change the solvent type or the stationary phase, you will change the relative retention. Sketch the chromatogram ifincreases 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 and k2are constant.

Short Answer

Expert verified

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

b) The sketch of chromatogram if increases 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, ifNandare 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

Retention factors for three solutes separated on aC8non-polar stationary phase are listed in the table. Eluent was a 70 : 30 (vol/vol) mixture of 50 mM citrate buffer (adjusted to pH withNH3) plus methanol. Draw the dominant species of each compound at each pH in the table and explain the behavior of the retention factors.

(a)explain how to measure k and resolution.

(b)state three method for measuring tmin reversed-phase chromatography.

(c)state three method for measuring tmin hydrophilic interaction liquid chromatography.

(d)Estimate tmfor 150.46column containing 5-渭尘particles operating at a flow rate of 1.5mL/minEstimate tmif the particle size were 3.5-渭尘 instead.

(a) Nonpolar aromatic compounds were separated by HPLC on an octadecyl(C18)bonded phase. The eluent was 65 vol% methanol in water. How would the retention times be affected if 90% methanol were used instead?

(b) Octanoic acid and 1-aminooctane were passed through the same column described in (a), using an eluent of 20% methanol/80% buffer (pH 3.0). State which compound is expected to be eluted first and why.

role="math" localid="1656416023291" CH3CH2CH2CH2CH2CH2CH2CO2HOctanoicacidCH3CH2CH2CH2CH2CH2CH2CH2NH21Aminooctane

(c) Polar solutes were separated by hydrophilic interaction chromatography (HILIC) with a strongly polar bonded phase. How would retention times be affected if eluent were changed from 80 vol% to 90 vol% acetonitrile in water?

(d) Polar solutes were separated by normal-phase chromatographyon bare silica using methyl t-butyl ether and 2-propanol solvent. How would retention times be affected if eluent were changed from 40 vol% to 60 vol% 2-propanol? (Hint: See Table 25-4.)

a. Why is high pressure needed in HPLC?

b. For a given column length , why do smaller particles give a higher plate number?

c. What is bonded phase in liquid chromatography?

(a). Sketch a graph of the van Deemnter equation (plate height versus linear flow rate).what would the curve look like if the multiple path term were 0? If the longtitundinal diffusinal diffusion term were 0?

(b). Explain why the van Deemter curve for 1.8渭尘particles in figure 25-3is nearly flat at high flow rate.what can you say about each of the terms in the van Deemeter equation for 1.8渭尘particles.

(c). Explain why the 2.7渭尘superficially porous particle enables separations similar to those achieved by 1.8渭尘totally porous particles,but the superficially porous particle requires lower pressure.

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