A High-Pressure Liquid Chromatographic Method for Analysis of Carbohydrates and Polyols from Lichens

Abstract

An analysis of 80% ethanol-soluble carbohydrates and polyols (polyhydric alcohols) from the lichens Ramalina stenospora Miill. Arg. and R. complanata (Sw.) Ach., was made by means of a high-pressure liquid chro- matographic method. Two chromatographic columns, utBondapak carbohy- drate and Aminex Q-150S, were used to effect the analysis. Of the two columns used the yBondapak column was found to resolve the larger number of carbohydrates from synthetic mixtures; however, the Aminex Q-150S column was required for resolution of mannitol-glucose-sorbitol and arabitol-ribitol complexes. Aminex Q-150S was more efficient in the separa- tion of polyols than monosaccharides. Utilization of high-pressure liquid chromatography was found to be a rap- id, sensitive method for analysis of carbohydrates and polyols routinely found in lichen extracts. It has the desirable features of small sample size, recover- able fractions, rapidity and ease of quantification of lichen metabolites. The assessment of carbohydrate and polyhydric alcohol content in lichens, as well as the biotrophic transport of these products, have become important parameters in the analyses of physiological processes in lichen symbionts. Several methods of anal- yses of carbohydrates and polyhydric alcohols (polyols) from lichens and fungi are in current use (Lewis & Smith, 1967; Culberson, 1969, 1970). Development of automated assay methods to determine these products in lichens has focused upon gas chroma- tography (Komiya & Shibata, 1971; Culberson, et al. 1977). The method of high-pressure liquid chromatography (HPLC), which is reported in this paper, has features of rapidity, sensitivity, resolution and fraction recovery, and eliminates the need of sample derivatization required in gas chromatography (Brown, 1973). HPLC was first applied to lichen metabolites to elucidate the derivatives of the