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Chapter 25: Q7P (page 707)
If along HPCL column has a place height of 5.0 what will be the half-width (in seconds) of a peak eluted at 10.0min? if plate height,what will be?
The half-width (in seconds) of a peak eluted.
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Why are silica stationary phase height limited to operating in the pH range ?why does the silica in figurehave improved stability at low pH?
(a) List ways in which the resolution between two closely spaced peaks might be changed.
(b) After optimization of an isocratic elution with several solvents, the resolution of two peaks is 1.2How might you improve the resolution without changing solvents or the kind of stationary phase?
Chromatography–mass spectrometry. HPLC separation of
enantiomers of the drug Ritalin on a chiral stationary phase was
shown in Problem 25-13.
(a) Detection is by atmospheric pressure chemical ionization with
selected reaction monitoring of the m/z 23484 transitions. Explain
how this detection works and propose structures for m/z 234 and m/z 84.
(b) For quantitative analysis, the internal standard-Ritalin with
a deuterated methyl group was added. Deuterated enantiomers have
the same retention times as unlabelled enantiomers. Which selected
reaction monitoring transition should be monitored to produce a
chromatogram of the internal standard in which unlabelled Ritalin
will be invisible?
Question: Explain how to use a gradient for the first run to decide whether isocratic or gradient elution would be more appropriate.
Question: Literature search problem: Human serum albumin (HSA) is an important protein ingredient in cryopreservation media used in procedures such as in vitro fertilization. Search the literature for a high-performance liquid chromatography method for the determination of human serum albumin and the stabilizer N-acetyl tryptophan in medical devices.
(a) Give the citation (authors, title, journal name, year, volume, pages) for the research paper that fits the criteria of this analysis.
(b) What alternative methods could be used for analysis of human serum albumin?
(c) What type of analytical column is used for the separation?
(d) How long was the gradient? How long were the additional wash and equilibration steps within the gradient method?
(e) What parameters were assessed in the method validation?
(f) Why were particles with 300 Å pores used?
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