A new method for the separation and identification of oligosaccharides obtained by enzymatic digestion of hyaluronic acid (HA) with hyaluronidase (EC 184.108.40.206) using on-line high-performance liquid chromatography/electrospray mass spectrometry (HPLC/ESI-MS) is presented. Reversed-phase ion pairing-HPLC, based on tributylamine salts and a volatile mobile phase, provided excellent chromatographic resolution and separation was achieved for HA oligosaccharides containing 2-40 monomers (from 2- to 40-mers). Using the on-line ion trap mass analyzer, complete identification and structural information for each HA oligomer species was obtained. In particular, a series of negatively charged species of different m/z ratios are seen for each oligosaccharide. Smaller HA species, from 2- to 4-mers, exhibit mainly [M-H](-1) anions, whereas the 6-10-mers exist predominantly as the charge state of -2. The HA oligomers from 12- to 18-mers are mainly represented by [M-3H](-3) anions while species from 20- to 28/30-mers are characterized by a charge state of -4. HA oligosaccharides from 32- to 40-mers exist as [M-5H](-5) anions. Furthermore, for smaller HA species, from 4/6- to 18/20-mers, ESI-MS revealed, generally in low relative abundance, anions related to the loss of one/two monosaccharide unit(s) from the oligomers, and no odd-numbered anions were produced for HA species greater than 20-mers.
|Data di pubblicazione:||2007|
|Titolo:||On-line HPLC/ESI-MS separation and characterization of hyaluronan oligosaccharides from 2-mers to 40-mers.|
|Codice identificativo ISI:||WOS:000248993300043|
|Codice identificativo Scopus:||2-s2.0-34548033741|
|Codice identificativo Pubmed:||17608452|
|Citazione:||On-line HPLC/ESI-MS separation and characterization of hyaluronan oligosaccharides from 2-mers to 40-mers / Volpi, Nicola. - In: ANALYTICAL CHEMISTRY. - ISSN 0003-2700. - STAMPA. - 79(16)(2007), pp. 6390-6397.|
|Tipologia||Articolo su rivista|
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