Abstract
Publisher Summary This chapter discusses different stationary phase matrices and functionalities available for the high-performance liquid chromatography (HPLC) separation of carbohydrates. Carbohydrates play an important role in many different research and industrial domains, such as biochemistry, clinical chemistry, biology, pharmacy, biotechnology, and food chemistry. Automated chromatographic analysis of carbohydrates is most frequently performed on ion-exchange stationary phases. The ion-exclusion phenomenon can be explained as the repulsion of ions with equidirectional charges. Spectrophotometric postcolumn labeling has played an important role in monitoring carbohydrates in borate complex anion-exchange chromatography because carbohydrates have no characteristic absorption in the ultraviolet (UV)-visible range. Measurement of the refractive index is insensitive and is prone to baseline drift because of temperature instability or changes in mobile-phase composition. The increase in retention times with increasing alkyl chain length as observed with linear acids, aldehydes, and alcohols is mainly caused by a reversed-phase mechanism. The elution of oligomeric analytes occurs in order of decreasing size. If there are no additional interactions between the analytes and the stationary phase, a linear correlation between the elution volume and the logarithm of molecular weight can be obtained.
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