Hydrolysis of bacterial wall carbohydrates in the microwave using trifluoroacetic acid

By and large, monosaccharide composition and linkage analyses of bacterial cell-surface carbohydrates are achieved by hydrolysis into the corresponding monomeric constituents, and characterization of these, or their derivatives, by chromatographic and spectrometric methods. Normally, these hydrolyses are carried out conveniently with trifluoroacetic acid (TFA) at high temperatures for long periods of time, for example, in 4 M TFA at 100 °C for 5 h in a heating block. In this study, using a closed-vessel system, we investigated the effectiveness and reliability of microwave-assisted TFA hydrolysis of bacterial lipopolysaccharides, capsule, and teichoic-acid polysaccharides that were variably composed of several glycoses. In all cases, we were able to establish that 5 min of hydrolysis in the microwave at 120 °C with 4 M TFA (measured pressure of 90 psi) was sufficient time to obtain comparable results to those afforded by conventional hydrolysis. The same observation was made when fully methylated carbohydrates were hydrolyzed. The data obtained with our microwave system (Aurora Instruments MW600) showed that microwave-induced hydrolysis can be used with a high degree of confidence to carry out sugar composition analysis of complex bacterial glycans in markedly shorter periods of time. The results also suggested that non-thermal mechanistic factors must also be involved, at least in part, in accelerating the reaction rate of glycosidic hydrolysis.

Microwave-accelerated hydrolysis of bacterial polysaccharides
• 5 h for conventional hydrolysis with 4 M TFA at 100 oC.
• 5 min for microwave hydrolysis with 4 M TFA at 120 oC (90 psi).Figure optionsDownload as PowerPoint slide