Structure elucidation of highly polar basic degradants by on-line hydrogen/deuterium exchange hydrophilic interaction chromatography coupled to tandem mass spectrometry.
A hydrophilic interaction liquid chromatography (HILIC) method was used to separate a commonly used pharmaceutical starting material, 4-aminomethylpyridine (4-AMP), and its degradants. The structures of the major degradants were characterized and elucidated without prior isolation by accurate mass measurement, MS/MS analysis and on-line hydrogen/deuterium (H/D) exchange experiments. The mass spectra obtained from H/D exchange experiments are particularly useful to differentiate structural isomers, to elucidate the fragmentation pathways, and to aid in structure elucidation in the absence of MS/MS fragmentation information. The impact of deuterium oxide and temperature on HILIC separation has also been explored here. The integration of H/D exchange with HILIC has been described here for the first time and has been demonstrated to be a powerful structure elucidation tool via the study of degradants in 4-AMP.
Liu
Min
M
Analytical Research & Development, Amgen, One Amgen Center Drive, Thousand Oaks, CA 91320, USA. mliu@amgen.com
Ronk
Mike
M
Ren
Da
D
Ostovic
Judy
J
Cauchon
Nina
N
Zhou
Zhaohui Sunny
ZS
Cheetham
Janet
J
D002853:Chromatography, Liquid / Q000379:methods*; D041961:Deuterium Exchange Measurement / Q000379:methods*; D017666:Deuterium Oxide / Q000737:chemistry*; D007539:Isonicotinic Acids / Q000737:chemistry; D009536:Niacinamide / Q000737:chemistry; D011725:Pyridines / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013696:Temperature
D007539:Isonicotinic Acids; D011725:Pyridines; D009536:Niacinamide; C027681:isonicotinamide; D017666:Deuterium Oxide