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Signal saturation for Waters 2475 fluorescence detector
Discussions about HPLC, CE, TLC, SFC, and other "liquid phase" separation techniques.
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I have a Waters Breeze I HPLC system with a Waters 2475 fluorescence detector connected to it through a SAT-IN module. I have a method set up to detect a mixture of compounds by fluorescence. I'm interested in the lower abundance compound. Unfortunately, there is a highly fluorescent compound present which saturates the detector. I don't want to decrease the sensitivity of the detector because I lose the signal form my compound of interest. I want to turn off the SAT-IN module while the highly fluorescent compound elutes, but I don't know how. The Breeze software has an "events" function for the SAT-IN module that toggles between "off and on" at times entered in by the user. I thought it corresponded to an enable/disable function for the module channel so that I could turn it off. I tried it, but nothing happens. Any suggestions?
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Turning off the satin module will not prevent saturation of your 2475 detector. Maybe you could use the timed events in the 2475 to use less a optimal exitation wavelenght as your highly fluorescent compound elutes and then switch back to the optimal exitation for your compoud of interest. You also might want to consider upgrading to Breeze 2 software as it has direct 2475 control.
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Why bother with the huge peak if it is separated from your analyte? If it is overlapping than no electronic reduction of the large peak will do any good, you will have to change the chromatography or chemistry.
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