Long analysis with multiple methods

[ScottHorn]

I would like to use Clarity to perform a very long affinity purification, and it seems that I need to use multiple methods throughout the run. The process consists of loading the sample, washing unbound proteins from the column, reversing flow direction, elution, washing, and column regeneration. Trying to control all these steps through one event table leads to conflicts between events that rely on the same detector for their starting conditions. I've thought about using a sequence, but without an autosampler. I think I could treat each step as a separate analysis in the sequence, and use a different method and event table for each. My problem is that I cannot figure out how to simulate the start signals from the autosampler in order to make Clarity advance from one step in the sequence to the next. Is it possible to use Clarity's digital outputs and inputs to control a sequence without an autosampler? Could I somehow have the sequence advance at the end of each run without waiting for start signals? Another method I'm curious about it using a batch file and command line parameters to run each step as a single analysis with a different method. Can you think of any solution to this problem?

Scott Horn

[Ivan Vins]

To use the sequence with different methods will be a suitable solution for this task.

In Clarity System configuration you specify the Start In digital input and Ready Out digital output. Sequence in active mode controls the Ready Out output - once it is started, it changes its level and waits for the Start In. When the Start signal is received, the level is changed back to prevent next injection till the sequence is ready for it. In case those outputs are interconnected directly, the analysis will start immediately. The Idle Time set in Sequence, Options may be needed to allow downloading the new method parameters to the controlled modules before the acquisition start.

In the current version 2.7 you could use the new Virtual Digital Input Output loop auxiliary module, which simulates such interconnection even if suitable digital outputs/inputs are not available on the hardware. It is even possible to set up a delay for the reaction in the Method setup, Aux tab.

[Honeybee]

I am interested in constructing the same kind of run as ScottHorn, e.i. a long affinity purification with several steps with a manual injector.
I am however not as versed as Scott in the digital language.
I am thinking that I am to have a new method for each of the following steps: equilibration, injection, wash and elution.
How am I to set up the methods in order for the sequence to work? All of the methods can be set to a specified time and do not have to rely on any signal from the detector.

Would it be possible to explain this for me?

[Ivan Vins]

The above application was described in some detail

https://theanalyticalscientist.com/app- ... tive-hplc/

The methods could be run using the Sequence table. The sequence will perform the methods automatically in order as specified in the table, starting a next one when the previous is finished. The needed settings are:

a loop between the Start IN digital input and the Ready OUT digital output specified in the System, Configuration window - could be created by physically connecting the respective leads on the A/D converter cable or by using the Virtual digital Input Output Loop auxiliary for this purpose.

Sequence options set to active sequence, with some Idle Time.

Please e-mail me to support@dataapex.com if more details are needed