Abstract
Procedures are described using high-performance liquid chromatography with pulsed amperometric detection for the measurement of neutral sugars and amino sugars released upon acid hydrolysis of non-starch polysaccharides. One procedure measures the neutral sugars with near-baseline separation in a 36 min run. The second procedure measures amino sugars and all neutral sugars except rhamnose and arabinose, which co-elute under the conditions described, in a 40 min run. Neutral sugars released from non-starch polysaccharides are ionized at high pH and separated by anion-exchange chromatography using NaOH as the eluent. When hexosamines or N-acetylhexosamines are present, as in mycoprotein and mushrooms or in ileostomy effluent, separation is achieved isocratically using 0.1 mmol dm–3 NaOH. The sulfate ions present in the hydrolysate are prevented from reaching the analytical column by a pre-column guard. Post-column addition of 300 mmol dm–3 NaOH increases the analytical signal and minimizes baseline drift. When only neutral sugars are present in the hydrolysate a single dilution step is required before analysis. When neutral sugars and hexosamines are present, a simple two-step neutralization and dilution are performed before injection.
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