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GC-MS analysis of hydrolyzed and derivatized enzyme assay mixtures. (A) Sample chromatograms showed two peaks at 17.4 and 17.9 min: retention times corresponding to those for derivatives of authentic GlcNAc and ManNAc, respectively. ManNAcA eluted together with ManNAc since the ManNAcA carboxyl group was reduced to an alcohol by sodium borodeuteride. (b) MS analysis at 17.9 min was shown to contain peaks for both ManNAc (marked with an asterisk) and ManNAcA (numbered peaks).
