Norihisa Takada

Isolation, Characterization, and Surfactant Properties of the Major Triterpenoid Glycosides from Unripe Tomato Fruits

Various triterpenoid glycosides were extracted from whole unripe tomato fruits ( Lycopersicon esculentum cv. Cedrico), using aqueous 70% (v/v) ethanol to study their surfactant properties. Cation-exchange chromatography using a Source 15S column and subsequent semipreparative HPLC using an XTerra RP18 were employed to purify individual triterpenoid glycosides from the extract. The structure of the purified compounds was established by mass spectrometry and nuclear magnetic resonance spectroscopy. The furostanol glycoside tomatoside A (749 mg/kg of DW) and the glycoalkaloids alpha-tomatine (196 mg/kg of DW) and esculeoside A (427 mg/kg of DW) were the major triterpenoid glycosides present. Furthermore, minor amounts of a new dehydrofurostanol glycoside, dehydrotomatoside, were found. The critical micelle concentrations of the major triterpenoid glycosides, alpha-tomatine, tomatoside A, and esculeoside A, were determined as 0.099, 0.144, and 0.412 g/L, respectively. The results show that tomatoside A, and not the more well-known alpha-tomatine, is the predominant triterpenoidal surfactant in unripe tomato fruits.

C22 isomerization in α-tomatine-to-esculeoside A conversion during tomato ripening is driven by C27 hydroxylation of triterpenoidal skeleton.

Compositional analysis by liquid chromatography/mass spectrometry of triterpenoid glycosides in different tomato cultivars, ripening stages, and parts of fruits showed that alpha-tomatine was generally most abundant in the flesh of the mature green stage, whereas esculeoside A was predominant in that of the red ripe stage. The sum of these glycoalkaloids was more or less constant, suggesting that alpha-tomatine is converted to esculeoside A during ripening. Besides various substitutions, the C22alphaN --> C22betaN isomerization is an important step in this transformation. By quantum chemical calculations it was shown that hydroxylation at C27 of the triterpenoidal skeleton is the driving force behind the isomerization. For the protonated form of the glycoalkaloid (predominant at the pH of tomato tissue), the C22betaN configuration becomes more favorable than that of C22alphaN, through the extra energy provided by the hydrogen bond between the protonated nitrogen and the lone pair of the oxygen of the C27-OH.