Document Detail

An ultrasensitive chemical method for polysialic acid analysis.
MedLine Citation:
PMID:  11555620     Owner:  NLM     Status:  MEDLINE    
An ultrasensitive method for analysis of polysialic acid (polySia) chains, using fluorescence-assisted high-performance liquid chromatography was developed. The new method is a substantial improvement of our earlier method in which the reducing terminal Sia residues of a homologous series of oligo/polySia hydrolytically released during derivatization reaction were simultaneously labeled with a fluorogenic reagent, 1,2-diamino-4,5-methylenedioxybenzene (DMB) in situ. We first studied extensively the stability of oligo/polySia in the acid (0.02 M trifluoracetic acid) used for 1,2-diamino-4,5-methylenedioxybenzene derivatization under various conditions of reaction time and temperature, analyzing the hydrolytic products by high-performance anion exchange chromatography with pulsed electrochemical detection (HPAEC-PED). Then we optimized the reaction conditions to minimize degradation of the parent polySia while maintaining high derivatization rate. Using a DNAPac PA-100 column rather than a MonoQ column, baseline resolution of polySia peaks up to DP 90 with a detection threshold of 1.4 femtomol per resolved peak was achieved. The new method was used to analyze the degree of polymerization of a polySia-containing glycopeptide fraction derived from embryonic chicken brain, and the results were compared with those obtained by HPAEC-PED.
S Inoue; S L Lin; Y C Lee; Y Inoue
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Glycobiology     Volume:  11     ISSN:  0959-6658     ISO Abbreviation:  Glycobiology     Publication Date:  2001 Sep 
Date Detail:
Created Date:  2001-09-13     Completed Date:  2001-12-07     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  9104124     Medline TA:  Glycobiology     Country:  England    
Other Details:
Languages:  eng     Pagination:  759-67     Citation Subset:  IM    
Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan, ROC.
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MeSH Terms
Anion Exchange Resins
Brain Chemistry
Chick Embryo
Chromatography, High Pressure Liquid / methods*
Chromatography, Ion Exchange / methods*
Sensitivity and Specificity
Sialic Acids / analysis*
Reg. No./Substance:
0/Anion Exchange Resins; 0/Sialic Acids; 0/polysialic acid

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