1260 Infinity Binary LC - System User Guide
59
Optimization of the Agilent 1260 Infinity Binary LC
3
How to Achieve Higher Resolution
80 Hz data rate compared with 20 Hz data rate results in improvements
in:
•
Peak Width:
– 30 %
•
Resolution:
+ 30 %
•
Peak capacity:
+ 40 %
•
Apparent column efficiency:
+ 70 %
Chromatographic Conditions
As already stated previously, chromatographic conditions depend on the
compounds to be analyzed. But here also, some rules of thumb exist.
•
Moderate flow rates should be used, but recent experiments have shown
that elevated flow rates can also be advantageous for improved
separation. For 4.6 mm inner diameter columns packed with
sub- 2- micron particles, a flow rate of 2 mL/min is recommended as a
starting value. For 2.1 mm inner diameter sub- 2- micron columns,
0.4 mL/min is a good starting point.
•
Moderate gradients should be used, for example, 2 – 5 % gradient
change per minute.
•
The column temperature is an additional parameter for optimization.
Temperature can influence a separation, and should not be overlooked.
Table 6
Relation between data rate and chromatographic performance
Data Rate
Peak Width
Resolution
Peak Capacity
80 Hz
0.300
2.25
60
40 Hz
0.329
2.05
55
20 Hz
0.416
1.71
45
10 Hz
0.666
1.17
29
5 Hz
1.236
0.67
16
