Suppose that an HPLC column produces Gaussian peaks. The detector measures absorbance at 254 nm. A sample containing equal moles of compounds A and B was injected into the column. Compound A E254=2.26×104M-1cm-1has a height h=128mm and a half-width w1/2=10.1mm. Compound B E254=168×104M-1cm-1has w1/2=7.6mm. What is the height of peak B in millimeters?

Short Answer

Expert verified

The height of peak B in millimetershB=126.45mm

Step by step solution

01

Step 1:

Since the sample containing equal moles of compounds A and B, the peak Ares are proportional to molar absorptivity:

PeakApeakB=2.26.1041.68.104=1.064.hA.w1/21.064.hB.w1/2

02

Step 2:

The height of peak B is:

hB=1.064.hA.w1/2.1.68.1041.064.w1/2.2.26.104hB=1.064.128mm.10.1mm.1.68.1041.064.76mm.2.26.104hB=126.45mmHere,thefinalresultishB=126.45mm

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Most popular questions from this chapter

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The antitumor drug gimatecan is available as nearly pure (S)-enantiomer. Neither pure (R)-enantiomer nor a racemic (equal) mixture of the two enantiomers is available. To measure small quantities of (R)-enantiomer in nearly pure (S)-gimatecan, a preparation was subjected to normal-phase chromatography on each of the enantiomers of a commercial, chiral stationary phase designated (S,S)- and (R,R)-DACH-DNB. Chromatography on the (R,R)-stationary phase gave a slightly asymmetric peak at tr 5 6.10 min with retention factor k 5 1.22. Chromatography on the (S,S)- stationary phase gave a slightly asymmetric peak at tr 5 6.96 min with k 5 1.50. With the (S,S) stationary phase, a small peak with 0.03% of the area of the main peak was observed at 6.10 min.

Chromatography of gimatecan on each enantiomer of a chiral stationary phase. Lower traces have enlarged vertical scale. [Data from E. Badaloni, W. Cabri, A. Ciogli, R. Deias, F. Gasparrini, F. Giorgi, A. Vigevani, and C. Villani, “Combination of HPLC ‘Inverted Chirality Columns Approach’ and MS/MS Detection for Extreme Enantiomeric Excess Determination Even in Absence of Reference Samples.” Anal. Chem. 2007, 79, 6013.]

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(d) The column provides N 5 6 800 plates. What would be the resolution between the two equal peaks in a racemic (equal) mixture of (R)- and (S)-gimatecan? If the peaks were symmetric, does this resolution provide baseline separation in which signal returns to baseline before the next peak begins?

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(a) What mobile phase composition provides greatest retention(k)for the components? Least retention? Coelution (equalk)of two components?

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(c) Would60%acetonitrile yield adequate resolution?

(d) Assuming Gaussian peaks, does the separation at60%acetonitrile have the attributes of a good separation?

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