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Chapter 26: Q20P (page 748)

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.

Short Answer

Expert verified

TheKav for transferrin isKav=0.16

Step by step solution

01

Define ferric citrate:

Ferric citrate or iron (III) citrate describes any complex compounds formed by binding to any few conjugate bases derived from citric acid and ferric ions.

02

Calculate the maximum values for ferritin, ferric citrate, and transferrin:

Consider the following:

Ferritin maximum:

Vo=22×0.65mL=14.3mL

Ferric citrate maximum:

Vm=84×0.65mL=54.6mL

Transferrin maximum:

Vt=32×0.65mL=20.8mL

03

Calculate Kv for transferrin:

Next calculate the Kavfor transferrin by the following equation

Kav=Vt-VoVm-VoKav=20.8-14.354.6-14.3Kav=0.16

The solution toKav for transferrin isKav=0.16

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Most popular questions from this chapter

In ion-exclusion chromatography, ions are separated from nonelectrolytes by an ion-exchange column. Nonelectrolytes penetrate the stationary phase, whereas ions of the same charge as the resin are repelled by the fixed charges. Because co-ions have access to less of the column volume, electrolytes are eluted before nonelectrolytes. The chromatogram shows the separation of trichloroacetic acid (TCA,pKa=-0.5), dichloroacetic acid (DCA,), and monochloroacetic acid (MCA,pKa=2.86) by passage through a cation-exchange resin eluted with 0.01 M HCl. Explain why the three acids are separated and why they emerge in the order shown.

26-D. Consider a capillary electrophoresis experiment conducted near pH9, at which the electroosmotic flow is stronger than the electrophoretic flow.

(a) Draw a picture of the capillary, showing the placement of the anode, cathode, injector, and detector. Show the direction of electroosmotic flow and the direction of electrophoretic flow of a cation and an anion. Show the direction of net flow.

(b) Using Table 15-1, explain why CI-has a shorter migration time than I-Predict whether Br-will have a shorter migration time than CI-or a greater migration time than I-

(c) Why is the mobility of I-greater than that ofCI-?

(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

Consider a negatively charged protein adsorbed on an anion-exchange gel at pH 8.

(a) How will a gradient of eluent pH (from pH 8to some lower pH) be useful for eluting the protein? Assume that the ionic strength of the eluent is kept constant.

(b) How would a gradient of ionic strength (at constant pH) be useful for eluting the protein?

State three different methods to reduce electroosmotic flow. Why does the direction of electroosmotic flow change when a silica capillary is washed with a cationic surfactant?
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