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Development of a method for dansylation of metabolites using organic solvent-compatible buffer systems for amine/phenol submetabolome analysis.
Hsieh, Ya-Ju; Chien, Kun-Yi; Chang, Chun-Ming; Hung, Cheng-Yu; Li, Liang; Chiang, Wei-Fan; Lee, Cheng-Chia; Chang, Chih-Hsiang; Chang, Ying-Hsu; Yu, Jau-Song; Chen, Yi-Ting.
Afiliación
  • Hsieh YJ; Molecular and Medicine Research Center, Chang Gung University, Taoyuan, Taiwan.
  • Chien KY; Molecular and Medicine Research Center, Chang Gung University, Taoyuan, Taiwan; Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan, Taiwan; Department of Cell and Molecular Biology, Chang Gung University, Taoyuan, Taiwan.
  • Chang CM; Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan, Taiwan.
  • Hung CY; Molecular and Medicine Research Center, Chang Gung University, Taoyuan, Taiwan.
  • Li L; Department of Chemistry, University of Alberta, Edmonton, AB, T6G2G2, Canada.
  • Chiang WF; Department of Oral and Maxillofacial Surgery, Chi-Mei Medical Center, Tainan, Taiwan; School of Dentistry, National Yang Ming University, Taipei, Taiwan.
  • Lee CC; Kidney Research Center, Department of Nephrology, LinKou Chang Gung Memorial Hospital, Taoyuan, Taiwan; Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
  • Chang CH; Kidney Research Center, Department of Nephrology, LinKou Chang Gung Memorial Hospital, Taoyuan, Taiwan; Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
  • Chang YH; Department of Urology, New Taipei Municipal Tu Cheng Hospital, Chang Gung Memorial Hospital and Chang Gung University, Taiwan.
  • Yu JS; Molecular and Medicine Research Center, Chang Gung University, Taoyuan, Taiwan; Department of Cell and Molecular Biology, Chang Gung University, Taoyuan, Taiwan; Liver Research Center, LinKou Chang Gung Memorial Hospital, Taoyuan, Taiwan; Research Center for Food and Cosmetic Safety, College of Huma
  • Chen YT; Molecular and Medicine Research Center, Chang Gung University, Taoyuan, Taiwan; Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan, Taiwan; Kidney Research Center, Department of Nephrology, LinKou Chang Gung Memorial Hospital, Taoyuan, Taiwan; Department of Biomedical Sciences
Anal Chim Acta ; 1189: 339218, 2022 Jan 02.
Article en En | MEDLINE | ID: mdl-34815039
Metabolomics, which serves as a readout of biological processes and diseases monitoring, is an informative research area for disease biomarker discovery and systems biology studies. In particular, reversed-phase liquid chromatography-mass spectrometry (RPLC-MS) has become a powerful and popular tool for metabolomics analysis, enabling the detection of most metabolites. Very polar and ionic metabolites, however, are less easily detected because of their poor retention in RP columns. Dansylation of metabolites simplifies the sub-metabolome analysis by reducing its complexity and increasing both hydrophobicity and ionization ability. However, the various metabolite concentrations in clinical samples have a wide dynamic range with highly individual variation in total metabolite amount, such as in saliva. The bicarbonate buffer typically used in dansylation labeling reactions induces solvent stratification, resulting in poor reproducibility, selective sample loss and an increase in false-determined metabolite peaks. In this study, we optimized the dansylation protocol for samples with wide concentration range of metabolites, utilizing diisopropylethylamine (DIPEA) or tri-ethylamine (TEA) in place of bicarbonate buffer, and presented the results of a systemic investigation of the influences of individual processes involved on the overall performance of the protocol. In addition to achieving high reproducibility, substitution of DIPEA or TEA buffer resulted in similar labeling efficiency of most metabolites and more efficient labeling of some metabolites with a higher pKa. With this improvement, compounds that are only present in samples in trace amounts can be detected, and more comprehensive metabolomics profiles can be acquired for biomarker discovery or pathway analysis, making it possible to analyze clinical samples with limited amounts of metabolites.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Fenol / Aminas Tipo de estudio: Guideline Idioma: En Revista: Anal Chim Acta Año: 2022 Tipo del documento: Article País de afiliación: Taiwán Pais de publicación: Países Bajos
Texto completo
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Imprimir
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Fenol / Aminas Tipo de estudio: Guideline Idioma: En Revista: Anal Chim Acta Año: 2022 Tipo del documento: Article País de afiliación: Taiwán Pais de publicación: Países Bajos