Purification of monoterpenyl glycosides by gel-permeation and hydrophobic-interaction chromatography on polyacrylamide (Bio-Gel P-2).

Hydrophobic interaction of the aglycone of monoterpenyl glycosides with the polyacrylamide matrix of Bio-Gel P-2 greatly retards the elution of these substances when chromatographed in dilute aqueous sodium chloride. This hydrophobic interaction is eliminated by inclusion of 15% acetonitrile in the eluant, thereby permitting conventional gel-permeation chromatography. Combination of these techniques by sequential chromatography on the same Bio-Gel column, in the hydrophobic interaction mode followed by the gel-permeation mode, provides a simple, yet mild and highly selective procedure for the purification of monoterpenyl glycosides from crude plant extracts. Examination of the chromatographic properties of beta-D-glucopyranosides and beta-D-galactopyranosides of a number of acyclic, monocyclic, and bicyclic monoterpenols indicates that the extent of hydrophobic interaction is of diagnostic value in determining the nature of the aglycone.

Metabolism of monoterpenes: lactonization of (+)-camphor and conversion of the corresponding hydroxy acid to the glucoside-glucose ester in sage (Salvia officinalis).

The bicyclic monoterpene ketone (+)-camphor is a major constituent (up to 26%) of the volatile oil of immature sage (Salvia officinalis L.) leaves; however, as the plant matures the content of this ketone declines in the fully expanded leaves (to about 65% of maximum) as does the overall yield of oil (to roughly 60% of maximum). Examination of the metabolism of (+)-[G-3H]camphor in discs prepared from mature leaves of flowering sage plants revealed that this ketone was converted to a water-soluble metabolite which on chromatographic analysis proved to be considerably more polar than a simple monoterpenyl glycoside. Mass spectral analysis of several derivatives of the terpenoid moiety of the metabolite obtained from large-scale incubations allowed identification of the aglycone, while degradative studies and detailed radiochromatographic analyses indicated that the metabolite contained two glucose residues; one glycosidically linked and the other in ester linkage. All of the evidence was consistent with the initial lactonization of camphor to 1,2-campholide followed by conversion to the beta-D-glucoside-6-O-glucose ester of the corresponding hydroxy acid (1-carboxymethyl-3-hydroxy-2,2,3-trimethyl cyclopentane). Direct evidence for the intermediacy of 1,2-campholide was also obtained through isotopic dilution experiments and by direct testing of the labeled lactone. The lactonization of camphor in sage resembles a similar step in the catabolism of camphor by microorganisms, but appears to be the first report of this reaction type in higher plants.