35
Troubleshooting
Fraction Collector-Related Problems
»
The collection arm does not position
correctly over each collection vessel:
»
Ensure that the racks and tray(s) are aligned correctly.
»
Ensure that the correct rack type has been selected in
the
Rack
panel for the rack in use (see page 10).
»
Ensure that there is nothing obstructing
or restricting the arm movement.
If this does not solve the problem, the collection arm may
need to be recalibrated. Contact Biotage 1-Point Support.
»
Dripping needle and/or inconsistent dispensing
volumes
can be signs of a dirty collect valve.
Please contact Biotage 1-Point Support.
Gradient Problems
Low composition gradient is not correct
To improve gradient accuracy at very low percentages, premix
solvents using the desired final % strong solvent in the weak
solvent and use this as solvent B. Program the gradient from
0 to 100% B using the pre-mixed solvent B.
The baseline drift is different from the programmed gradient
Two factors contribute to altering the gradient as observed by
the UV detector:
1. As it takes at least 1 CV for the solvent to pass through the
column and reach the UV detector, the initial front of the
gradient will always be delayed by at least 1 CV compared to
the programmed gradient. A longer gradient delay may be
due to interactions with the silica, where strong solvent is
being selectively retained on the column.
2. The gradient as observed by the UV detector will at times
decline and plateau before the programmed gradient
does. This is due to the detection limit of the detector.
Any increase in the concentration of strong solvent will not
be registered as no light is reaching the detector at those
particular wavelengths.
If using a system with a Spektra software license, try performing
the run with the
UV Baseline Correction
option turned on.
Non-linear behavior due to
detection limit of the detector
Longer delay than expected (approx. 2 CV instead of 1 CV)
due to interactions with the silica, where strong solvent is
being selectively retained on the column
Figure 68.
Problems that can be seen in the UV signal.
UV Detector-Related Problems
»
No signal.
Check that the UV flow cell is correctly
mounted, see step 7 in “Clean the Flow Cell
of the UV Detector” on page 30.
»
No light (“UV detector error. Error code 2,30”).
Clean the flow cell; see page 30.
»
Noise
may be due to a contaminated flow cell.
Clean the flow cell; see page 30.
»
Drifting baseline
may be due to:
»
Contaminated flow cell. Clean the
flow cell; see page 30.
»
Used solvent is absorbing light at the selected
wavelength(s). Change the collection and fractionation
wavelength(s) or turn on the
UV Baseline Correction
option (only on systems with a Spektra software license).
»
Defective UV lamp. Contact Biotage 1-Point Support.
»
UV baseline correction is not eliminating drifting baseline.
The gradient is modified by the user during the run. If the
gradient is changed outside of its original boundaries, this
is not covered by the standard UV baseline correction (start
to end mix A/B). Change to full UV baseline correction (start
mix to 100% B) in the system settings (see page 24).
»
Missing peaks when using UV baseline correction.
If expected peaks do not show when you have the
UV Baseline Correction
option turned on and you are
using solvents that are high-absorbing over a wide
range of wavelengths (e.g. acetone or toluene), try
performing the run without UV baseline correction.
Troubleshooting
Summary of Contents for Selekt Enkel
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