In the figure below is a liquid chromatogram for four deoxymononucleotides: dAMP (deoxyadenosine monophosphate), dCMP (deoxycytidine monophosphate), dGMP (deoxyguanosine monophosphate), and dTMP (deoxythymidine monophosphate).
0.00E02
1
7.0002
3
6.001.02
5.000.02 ne 4000-02
1.00102
2.00-02
1001-02
0.000-00
1.000-02
100 200
30
400
100
700
imei
Details of the separation:
Column length: 25.0 cm
Column diameter: 0.46 cm
Stationary phase: C-18
Mobile phase: aqueous buffered at pH 7.2
Flow rate: 1.2 mL/min
Dead time: 16.7 s
Elution order: first to last - dAMP, dCMP, dGMP, dTMP
Quantity Component Peak height (au) 0.072 Elution me 120 Peak width at bases 28
Component Component 2 3 0.046 0.061 0.041 255 310 608 70 55 98
Questions:
1) What is the retention factor for dAMP?
2) What is the column efficiency for dAMP?
3) Assume the average theoretical plate height is 0.08 cm. At what approximate distance traveled in the column after injection of the sample were CMP and dMP separated with a resolution of 1.5, assuming the rest of the experiment remains constant?
4) Assume the average theoretical plate height is 0.08 cm. What column length would be needed to separate dCMP and dGMP with a resolution of 2.0, assuming the rest of the experiment remains constant?