DropSync on: When using DropSync the sensors in the Tube sensor detect when a drop is released.
The Tube rack moves and positions the next tube under the Tube sensor just after a drop is released.
Fractionation with DropSync can be used at flow rates up to 2 ml/min. For water and solutions with
higher surface tension, a higher flow rate can be used. Volatile solutions and solutions with low surface
tension may require a lower flow.
Missing tubes in Fraction collector F9-R
If a tube is missing, Fraction collector F9-R will continue the fractionation on the tube row located
closer to the center of the fraction collector. The fractionation marks in the chromatogram will then
not reflect the tubes in which the sample is collected.
If the fraction collector runs out of tubes, the delivery arm moves to the fraction collector center position
while ÄKTA pure pauses and displays an error message.
Missing tubes or plates in Fraction collector F9-C
When automatic cassette configuration is selected in the system settings the fraction collector auto-
matically detects which types of cassettes and plates that are present. The fraction collector will
however not detect if tubes or bottles are missing in the cassettes. Make sure that the cassettes to be
used are occupied by appropriate types and numbers of tubes or bottles before starting a run.
It is not possible to change the cassette configuration during a run. When the system state is set to
Pause it is possible to take out cassettes or plates from the fraction collector only if they are replaced
by cassettes or plates of the same type and are placed in the same positions.
The action of the system when the last tube in the fraction collector is filled is set in the instruction
Last tube filled in the system settings. The flow can be directed to waste or to any of the outlets or
the run can be paused. If the action is set to Pause, the system automatically pauses when the last
tube is filled and prompts the user to replace the filled tubes.
Peak broadening
The width of peaks at the fraction collector is influenced by the properties of the column and the di-
mensions of tubing connecting the components. Initial sample volume affects the peak width in gel
filtration (GF) chromatography. A sample zone is broadened during passage through a GF column so
that the sample is diluted and the resolution decreases with increasing sample volume. Sample volume
does not however affect the resolution in adsorption chromatography techniques such as affinity
chromatography (AC), ion exchange chromatography (IEX), and hydrophobic interaction chromatography
(HIC). The effect of peak broadening in the system from sample injection to peak detection (including
dilution on the column) is apparent in the chromatogram from the UV monitor, but broadening from
the UV monitor to fraction collection is not visible in the chromatogram. This "hidden" effect can
sometimes be dramatic, especially for high-resolution columns.
To minimize peak broadening, use narrow and short tubing connections as far as possible. Remember
that using narrow tubing will increase the back pressure in the system.
210 ÄKTA pure User Manual 29119969 AB
5 Operation
5.8 Fractionation
5.8.3 Fractionation overview
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