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Optimum split flow in ESI LC-MS

Posted: Thu Oct 26, 2006 10:10 am
by Rob Burgess
Working in the pharma. industry the majority of our analytes are basic, and thus we tend to work in the +ve ESI mode when using LC-MS quadropole instruments.

Recently we have been having trouble with obtaining suitable S/N sensitivity to identify unknown peaks in our total ion chromatograms (TIC). We have only been observing the main peak. Our analytical flow rate is 1.0 ml/min and we run with water/AcN/MeOH gradients that contain 0.05% TFA. The split post column from the DAD is approximately set so that a flow of 500µL/min enters the LC-MS. Recently some colleagues, looking to optimise the LC-MS TIC response, reduced the effluent flow to the LC-MS to ca. 200µL/min. They observed a dramatic increase in the S/N ratio and were able to decipher more peaks than just the main peak!

My question then is: what is the optimum flow rate setting for the LC-MS in ESI +ve mode (quadrople) to obtain good S/N of small impurity peaks in the TIC trace? Is 200µL/min still too large? Also what flow splitter device would users recommend? Currently we are using a simple tee-piece but may not be most user friendly device out there. We are considering either a metered valve (http://www.upchurch.com/Products/specsh ... 70&vFrom=L) or a set flow splitter (http://www1.dionex.com/en-us/webdocs/43 ... ro_V12.pdf - p.6). Any opinions on such devices? It appears that the flow splitting box may have a set split flow ratio that may not be ideal if we ever wanted to chnage this.

Posted: Thu Oct 26, 2006 1:55 pm
by Noser222
It all depends. I've run a method with a flow rate of 0.6 mL/min and my sensitivity was just as good as at 0.2 mL/min.
Before I bought anything, I would run flow injection analysis at those lower flow rates to see if the sensitivity is any better.

Posted: Thu Oct 26, 2006 5:36 pm
by promochrom
Many people just connect the outlet of DAD with MSD to allow the whole portion of the sample enter the MSD (unless you need to do fraction collection). They use a 2-mm column to get a flow rate favourable for MSD (0.2 ml/min).

When you change the flow rate, parameters that affect ionization (such as N2 flow rate and temperature) may also need optimzation (just a reminder).