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HPLC Simulator

The document provides instructions for an HPLC simulator assignment due November 20th. Students are asked to run 5 experiments using different solvents and conditions to separate 5 compounds, and report their results including an explanation of why their conditions worked best. They are also asked to investigate the effects of temperature on one run and calculate resolution between two barely separated peaks using three different methods.

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Erin Love
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203 views1 page

HPLC Simulator

The document provides instructions for an HPLC simulator assignment due November 20th. Students are asked to run 5 experiments using different solvents and conditions to separate 5 compounds, and report their results including an explanation of why their conditions worked best. They are also asked to investigate the effects of temperature on one run and calculate resolution between two barely separated peaks using three different methods.

Uploaded by

Erin Love
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as DOCX, PDF, TXT or read online on Scribd
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Chem 325

HPLC Simulator
20, by noon

Fall 2015
Due: November

Instructions: Download (or run online) HPLC simulator @ hplcsimulator.org

Initial Settings:
Column = Agilent Zorbax SB-C18
Flow rate = 1 mL/min
Temperature = 25C.
Your goal: to separate the five compounds listed in the table on the lower right with the best
results (all five peaks resolved and k in the range 2-10 if possible). Report your conditions, with
an explanation, for the following five runs (no credit will be given without an appropriate
explanation). Suitable explanations will include why the particular condition worked best, and
comparison to other conditions (i.e. why is the %B different for ACN vs. MeOH?).
1) Use water (A) /methanol (B) and isocratic elution (vary % B via a sliding bar on top left)
2) Use water (A) /acetonitrile (B) and isocratic elution (vary % B)
3) Use water (A) /methanol or acetonitrile (B) and gradient elution (vary % B over some
time, play with time settings)
4) For one of the isocratic runs investigate the effects of temperature (sliding bar on middle
left)
Record typical HETP, N, k (range)
Lastly, choose a setting where two (any) peaks elute close together and are barely separated.
Using the data for these two peaks, calculate resolution using THREE methods (see eq-n
below). Hint: you have to estimate (measure) the width of the peak (w) which is best done by
zooming into the chromatogram.

Rs

1 k 2'
(tr )
N


1 k a' v e
w a v e 4 - 1

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