A SIMPLE HPLC METHOD FOR THE ISOLATION AND QUANTIFICATION OF THE ALLERGENS TULIPOSIDE-A AND TULIPALIN-A IN ALSTROEMERIA

A practical, rapid, reliable and sensitive method for the isolation and determination of the allergens tuliposide A and alpha-methylene-gamma-butyrolactone (tulipalin A), by reversed-phase high-performance liquid chromatography (RP-HPLC), has been developed in order to select Alstroemeria species for breeding purposes. From the aqueous extracts of flowers, stems and leaves, of several Alstroemeria species, the contents of 6-tuliposide A and tulipalin A were determined by isocratic RP-HPLC, using distilled water as mobile phase. The compounds were detected by an UV detector at 208 nm. Differences in 6-tuliposide A and tulipalin A content were found among the species investigated, with the highest concentrations in stems and flowers. The absence of other tuliposides (e.g., 1-tuliposide A, 1- and 6-tuliposide B) in extracts was proven by TLC, RP-HPLC, H-1- and C-13-NMR. 6-Tuliposide A and tulipalin A were identified by H-1- and C-13-NMR and comparison with authentic material, respectively. With this HPLC method, it is possible to investigate a large number of plants for their contents of tuliposide A and tulipalin A, within a minimum of time, and to isolate them directly from aqueous extracts.