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3-Amino-1-phenyl-2-pyrazoline-5-ketone as a heterobifunctional chromogenic reagent to derivatize reducing glycans for subsequent online LC/MS analysis.
Lu, Yu; Wang, Chengjian; Liu, Rendan; Jin, Wanjun; Wen, Yanan; Huang, Linjuan; Wang, Zhongfu.
Afiliación
  • Lu Y; Educational Ministry Key Laboratory of Resource Biology and Biotechnology in Western China, The College of Life Sciences, Northwest University, Xi'an 710069, China.
  • Wang C; Educational Ministry Key Laboratory of Resource Biology and Biotechnology in Western China, The College of Life Sciences, Northwest University, Xi'an 710069, China.
  • Liu R; Educational Ministry Key Laboratory of Resource Biology and Biotechnology in Western China, The College of Life Sciences, Northwest University, Xi'an 710069, China.
  • Jin W; Educational Ministry Key Laboratory of Resource Biology and Biotechnology in Western China, The College of Life Sciences, Northwest University, Xi'an 710069, China.
  • Wen Y; Educational Ministry Key Laboratory of Resource Biology and Biotechnology in Western China, The College of Life Sciences, Northwest University, Xi'an 710069, China.
  • Huang L; Educational Ministry Key Laboratory of Resource Biology and Biotechnology in Western China, The College of Life Sciences, Northwest University, Xi'an 710069, China. Electronic address: huanglj@nwu.edu.cn.
  • Wang Z; Educational Ministry Key Laboratory of Resource Biology and Biotechnology in Western China, The College of Life Sciences, Northwest University, Xi'an 710069, China. Electronic address: wangzhf@nwu.edu.cn.
Anal Biochem ; 549: 1-11, 2018 05 15.
Article en En | MEDLINE | ID: mdl-29524379
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 amino 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 amination 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 derivatized by different reaction mechanisms, having great importance for functional glycomics studies.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Polisacáridos / Espectrometría de Masa por Ionización de Electrospray / Pirazolonas Límite: Humans Idioma: En Revista: Anal Biochem Año: 2018 Tipo del documento: Article País de afiliación: China Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Polisacáridos / Espectrometría de Masa por Ionización de Electrospray / Pirazolonas Límite: Humans Idioma: En Revista: Anal Biochem Año: 2018 Tipo del documento: Article País de afiliación: China Pais de publicación: Estados Unidos