Chromatography Forum

I had following gradient and column for ATP detection.

Colum: waterxterra,C8,2.1x50mm.
5micron

Mobile Phase A:10mM DMHA( made by mixing 1.73 ml of DMHA(mw:129) in 998ml water) Adusted the Ph to 7.0 by dil HCl.
Mobile phase B: Methanol

ATP was made in water 50uM

I see ion at 130(ESI+VE) and 295(ESI+VE) all over the run more than 10 million count. nothing else. UV-VIS shows ATP comes out at 1.5 min. but dont see any in ion at 508 or neg 506 in -ve mode.

What happed to mass spec ? is 130 is due to DMHA..i dont understand what is 295 ?

Please help

Hello,

sorry for seeing this so late.

You might have degradation of phosphate moeities on heated steel surface and subsequent leakage. I have not checked how this is linked to the 295 m/z signal.

Suppression effects of carbonate on the interaction between stainless steel and phosphate groups of phosphate compounds in high-performance liquid chromatography and electrospray ionization mass spectrometry
Asakawa Y et al
Journal of Chromatography A
Volumes 1198-1199, 11 July 2008, Pages 80-86