3 - UPLC_Work_Practical_Considerations
3 - UPLC_Work_Practical_Considerations
3 - UPLC_Work_Practical_Considerations
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Running U-HPLC: <strong>Practical</strong> <strong>Considerations</strong><br />
Solvent Composition<br />
at Mixer<br />
Solvent Composition<br />
at Column Head<br />
System Delay (Dwell) Volume<br />
Timing Offset<br />
0<br />
Injection<br />
Actual mobile phase<br />
profile on original<br />
system measured at<br />
the column inlet<br />
System volume creates an offset before the solvent<br />
composition change reaches the inlet of column (i.e. an<br />
“isocratic hold” at the beginning of every gradient)<br />
Smaller Volume<br />
System Volume 0.35 mL<br />
{<br />
}<br />
Target System with smaller volume<br />
(less isocratic hold time)<br />
t g<br />
Shula Levin, Waters Israel<br />
x<br />
Time<br />
Different System Volumes<br />
Effect on Separation<br />
Original<br />
Instrument<br />
System Volume 0.0.7 mL<br />
Larger Volume<br />
System Volume 1.4 mL<br />
Target System with larger volume<br />
(longer isocratic hold time)<br />
12
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