Qualitative and quantitative analysis of small amine molecules by MALDI-TOF mass spectrometry through charge derivatization.
A pair of isotopically coded light and heavy reagents, tris(2,4,6-trimethoxyphenyl)phosphonium acetic acid N-hydroxysuccinimide esters (1 and 2) were synthesized and used to derivatize low molecular weight (<500 Da) molecules containing primary or secondary amine functional groups for MALDI-TOF MS analysis. The light and heavy reagents added a 573 and 600 Da positively charged tag to each analyte, respectively. In the presence of 10 times excess of tag reagents, the coupling reactions reached near completion within 10 min. The derivatization greatly facilitated MALDI analysis of small molecules and significantly improved the sensitivity of analysis, allowing a limit of detection in the low femtomole range. Additionally, the reaction mixtures were directly analyzed by MALDI without sample cleanup. The quantification of small molecules by MALDI-TOF MS was successfully achieved by analysis of isotopically coded light and heavy derivatives. MALDI-TOF quantitative analysis of a mixture of antibiotics yielded calibration curves in the concentration range from 0.3 to 30 pmol/microL with r2 values greater than 0.9995.
Lee
Peter J
PJ
Life Science Chemistry, Waters Corporation, 34 Maple Street, Milford, Massachusetts 01757, USA.
Chen
Weibin
W
Gebler
John C
JC
D000588:Amines / Q000032:analysis*; D000900:Anti-Bacterial Agents / Q000032:analysis; D002138:Calibration; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D000588:Amines; D000900:Anti-Bacterial Agents