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Rapid and sensitive chromatographic determination of free sialic acid in complex bio-pharma fermentation media samples

Citation

Alwael, H and Connolly, D and Paull, B, Rapid and sensitive chromatographic determination of free sialic acid in complex bio-pharma fermentation media samples, Analytical Methods, 4, (9) pp. 2668-2673. ISSN 1759-9660 (2012) [Refereed Article]


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Copyright Statement

Copyright 2012 Royal Society of Chemistry

DOI: doi:10.1039/c2ay25299f

Abstract

This paper describes a rapid, sensitive and reproducible liquid chromatographic method specifically for the quantitative determination of total free sialic acid, employing thiobarbituric acid as pre-column tagging agent, following the oxidation of sialic acid with periodic acid. The derivatised sialic acid was separated from 2-deoxy-d-ribose and the other components utilising a short C18 monolithic column, with total run-times of under 90 s per sample. The method was successfully applied to quantify sialic acid in a range of complex samples, e.g. yeastolate powders, basal media and in-process samples supplied from the biopharmaceutical industry. The selective method was highly reproducible (%RSD for method (n = 6) = < 2, for peak area (n = 6) = 1.3, for retention time (n = 6) = 0.6), linear (R2 = 0.9994, range 0.25-25 μmol L-1) and sensitive (LOD = 842 fmol based upon a 100 μL inj. vol.).

Item Details

Item Type:Refereed Article
Research Division:Chemical Sciences
Research Group:Analytical Chemistry
Research Field:Separation Science
Objective Division:Expanding Knowledge
Objective Group:Expanding Knowledge
Objective Field:Expanding Knowledge in the Chemical Sciences
Author:Paull, B (Professor Brett Paull)
ID Code:81020
Year Published:2012
Web of Science® Times Cited:2
Deposited By:Austn Centre for Research in Separation Science
Deposited On:2012-11-21
Last Modified:2016-10-26
Downloads:3 View Download Statistics

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