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negative mode direct infusion
Posted: Wed Apr 15, 2015 6:36 am
I've been doing a lot of negative mode direct infusion on an orbitrap lately. Almost always, the S/N level is awful even if I'm infusing high concentrations, e.g. 100 ÂµM. At first I thought it was because the samples contained some salt but recently I infused a sample which I'm sure did not contain any salt and still got very poor S/N. I haven't had the same trouble with positive mode. I'm wondering if it's because the analyte is not ionizing well due to the lack of anything to extract protons? Do people ever add base to improve the signal in negative mode? If so, what do people add and at what concentration?
Posted: Wed Apr 15, 2015 2:36 pm
Are you doing this at regular flow rates with the HESI source?
Posted: Wed Apr 15, 2015 4:33 pm
I'm infusing at 3 mL/min with the ESI source. This seems to work fine for positive mode or calibrant solution in negative mode.
Posted: Fri Apr 17, 2015 1:49 pm
Back when I was doing more negative mode direct infusion, I tested a ton of different spray solutions and found that a mix of 50:50 (vol/vol) 5 mM NaOH:2-propanol seemed to work best. Here's a link that describes a bit more of the methods in case it's helpful http://www.ncbi.nlm.nih.gov/pubmed/22290482
Posted: Sun Apr 19, 2015 2:00 pm
Hey Craig, I have found 50:50 water:ACN with 5mM piperidine works pretty well. This paper might be useful for you, too. http://www.ncbi.nlm.nih.gov/pubmed/22006635