Buffer loading for counteracting metal salt-induced signal suppression in electrospray ionization.
The decrease in the sensitivity of electrospray ionization mass spectrometry caused by the presence of metal salts, such as sodium chloride, in the sample matrix is well known and is particularly problematic for biological samples. We report here that addition of high levels of ammonium acetate can improve analyte signal in aqueous electrospray solutions and counteracts the signal suppression caused by sodium chloride. A approximately 3-fold improvement in S/N is obtained by adding 8 M ammonium acetate to aqueous solutions of cytochrome c without added sodium chloride. No organic solvents or acids are added into the electrospray solutions. The signal-to-noise ratios of cytochrome c and ubiquitin (10(-)(5) M) ions formed from aqueous solutions containing 2.0 x 10(-)(2) M sodium chloride are improved by factors of approximately 7 and 11, respectively, by adding 7 M ammonium acetate to the solution. We propose that this effect is a result of the precipitation of Na(+) and Cl(-) from solution within the evaporating electrospray droplets prior to the formation of gas-phase protein ions. This method is potentially useful for improving the abundance of protein ions formed from solutions in which the molecules have a nativelike conformation and is particularly advantageous for such solutions that have high levels of sodium.
Iavarone
Anthony T
AT
Department of Chemistry, University of California, Berkeley, California 94720-1460, USA.
Udekwu
Osita A
OA
Williams
Evan R
ER
D000085:Acetates / Q000737:chemistry* / Q000494:pharmacology; D002021:Buffers; D045304:Cytochromes c / Q000032:analysis; D012815:Signal Processing, Computer-Assisted; D012965:Sodium Chloride / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D025801:Ubiquitin / Q000032:analysis
D000085:Acetates; D002021:Buffers; D025801:Ubiquitin; D012965:Sodium Chloride; D045304:Cytochromes c; C018824:ammonium acetate