期刊
ANALYTICAL BIOCHEMISTRY
卷 549, 期 -, 页码 1-11出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2018.02.029
关键词
Heterobifunctional chromogenic reagent; 3-Amino-1-phenyl-2-pyrazoline-5-ketone (PAP); Reducing glycan; ESI-MS; LC-MS
资金
- National Natural Science Foundation of China [31670808, 31600647, 31370804, 21375103, 31300678]
- Natural Science Special Fund of Shanxi Provincial Education Department [16JK1782]
- Scientific Research Foundation of Northwest University [15NW18]
Sensitive analysis of glycans by liquid chromatography/mass spectrometry is significantly hampered by the lack of chromogenic or fluorescent groups on the glycan structures, as well as, their poor ionization properties In the present, a heterobifunctional chromogenic reagent 3-amino-1-phenyl-2-pyrazoline-5-ketone (PAP) bearing ammo and active methylene groups, which readily reacts with reducing glycans, was used for detection of the pre-column-labeled glycans via high-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS). The PAP derivatives with active methylene and amino groups were obtained via reductive animation in acidic medium and condensation of an active PAP methylene group with the reducing end of glycans in alkaline medium, respectively, and the PAP derivatives could be further functionalized, e. g., via glycan microarray preparation The conditions for the two reaction modes were optimized, the HPLC separation method of PAP derivatives was investigated, and the PAP derivatives of some glycans derived from biological samples were obtained and analyzed by ESI-MS and LC-MS Using this new reagent, reducing glycans can be selectively denvatized by different reaction mechanisms, having great importance for functional glycomics studies.
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