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Chapter 26: Q23P (page 748)

A polystyrene resin molecular exclusion HPLC column has a diameter of 7.8 mm and a length of 30 cm. The solid portions of the gel particles occupy 20% of the volume, the pores occupy 40%, and the volume between particles occupies 40%. (a) At what volume would totally excluded molecules be expected to emerge? (b) At what volume would the smallest molecules be expected? (c) A mixture of polyethylene glycols of various molecular masses is eluted between 23 and 27 ml. What does this imply about the retention mechanism for these solutes on the column?

Short Answer

Expert verified

(a) The volume at which totally excluded molecules be expected to emerge isVx=0.4脳14.3mL=5.7mL

(b) The volume at which the smallest molecules be expected is

Vy=0.8脳14.3mL=11.5mL.

(c) The solutes given in the task are adsorbed on the resin, if they weren鈥檛 they would be eluted at lower volumes.

Step by step solution

01

Define molecular exclusion:

Separation of chemical substances and particles by separate movements using a two-stage analytical system.

02

The volume at which totally excluded molecules be expected to emerge:

(a)

Here we will calculate the volume at which would the totally excluded molecules be expected to emerge.

Total volume of the column:

V=蟺谤2lV=蟺0.39cm2脳30cmV=14.3cm3=14.3mL

Final volume:

Note that r=diameter /2 which is 7.8 mm/2= 39 mm.

03

The smallest molecules be expected:

(b)

Calculate the volume at which the smallest molecules would be expected:

Vy=0.8脳14.3mL=11.5mL

04

The retention mechanism for these solutes on the column:

(c)

Answer the question about the retention mechanism for solutes given in the task on the column:

Solutes given in the task are adsorbed on the resin, if they weren鈥檛 they would be eluted at lower volumes.

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Most popular questions from this chapter

A gel filtration column has a radius, y.of 0.80 cm and a length,l, of 20.0 cm

(a).Calculate the volume,Vtof the column, which is equal to蟺谤2

(b). The interstitial volumeV0was 18.1 mL and the total volume of mobile phase was 35.8 mL.

Find.Kavfor a solute eluted at 27.4 mL.

(a) The suppressor in Figure 26-9 enables low parts-per-billion conductivity detection for anions such asCl-andBr-,.but very poor detection limits for anions such asCN-and borate. Explain why.

(b) Mixtures of sodium carbonate and bicarbonate can be used as eluent in suppressed-ion anion chromatography. Detection limits are poorer than with hydroxide eluent due to higher background conductivity. Explain why

Figure 26-24 shows the effect on resolution of increasing voltage from 28 to120kV

(a) What is the expected ratio of migration times(t120kV/t28kV)in the two experiments? Measure the migration times for peak 1 and find the observed ratio.

(b) What is the expected ratio of plates (N120kv/N28kv)in the two experiments?

(c) What is the expected ratio of bandwidths(蟽120kv/蟽28kv)?

(d) What is the physical reason why increasing voltage decreases bandwidth and increases resolution?

Ferritin (molecular mass 450 000), transferrin (molecular mass 80 000), and ferric citrate were separated by molecular exclusion chromatography on Bio-Gel P-300. The column had a length of 37 cm and a 1.5-cm diameter. Eluate fractions of 0.65 mL were collected. The maximum of each peak came at the following fractions: ferritin, 22; transferrin, 32; and ferric citrate, 84. (That is, the ferritin peak came at an elution volume of 22 3 0.65 5 14.3 mL)Assuming that ferritin is eluted at the interstitial volume and that ferric citrate is eluted at Vm, findKavfor transferrin.

State the effects of increasing cross-linking on an ion-exchange column.
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