Triterpenoid Content of Berries and Leaves of Bilberry Vaccinium myrtillus from Finland and Poland

Abstract

Triterpenoid compounds found in free and ester forms in extracts of entire fruits and leaves and in fruit and leaf cuticular waxes of bilberry (Vaccinium myrtillus L.) collected in Finland and Poland were identified and quantitated by gas chromatography-mass spectrometry coupled to a flame ionization detector (GC-MS/FID). The main bilberry triterpenoid profile consisted of α- and β-amyrin, α- and β-amyrenone, campesterol, cholesterol, citrostadienol (in berries), cycloartanol, erythrodiol, lupeol, 24-methylenecycloartanol, sitosterol, sitostanol, stigmasterol, stigmasta-3,5-dien-7-one, uvaol, oleanolic and ursolic aldehydes, and oleanolic, ursolic, 2α-hydroxyoleanolic, and 2α-hydroxyursolic acids. Friedelin and D:A-friedooleanan-3β-ol were found only in Finnish plants, whereas D:C-friedours-7-en-3β-ol and taraxasterol were found only in Polish plants. To our knowledge, this is the first thorough description of triterpenoid compounds in this species. The presented results revealed that the triterpenoid profile of bilberry varied considerably between different organs of the plant, regardless of the plant origin, as well as between plant samples obtained from the two geographical locations.

Triterpenoid profile of fruit and leaf cuticular waxes of edible honeysuckle Lonicera caerulea var. kamtschatica

“…Therefore, the presence and diversity of triterpenoids in cuticular waxes may be of great systematic significance, even if in some species these compounds are present only in trace amounts. The fruit cuticular wax of edible honeysuckle contains a strikingly low amount of triterpenoids (only 6.25% of the total wax extract), which is a feature that clearly distinguishes this berry (a member of the Caprifoliaceae) from edible berries of plants belonging to other families, e.g., bilberry Vaccinium myrtillus L. [20], cowberry V. vitis-idaea L. [19,28], and cranberry V. macrocarpon Ait. of the Ericaceae [29] or grapevine Vitis vinifera L. of Vitaceae [21,22].…”

“…The relative abundance of triterpenoids in cuticular waxes can also have other implications concerning the form in which these compounds occur in the plant. Ericaceae species accumulate significant amounts of tritepenoids in a free and esterified form in cuticular waxes, while they synthesize very small amounts of their glycoside conjugates called saponins found in other tissues [20,28,31]. In contrast, many plants belonging to the genus Lonicera are known for high levels of saponins, mainly of oleanolic acid and hederagenin [1], or lupane-type triterpenes as aglycones [32].…”

“…Extracts were fractionated by adsoption chromatography as described previously [19][20][21][22] on silica gel plates in the solvent system CHCl 3 /MeOH (97:3, v/v). Fractions were localized on plates by comparison with standards (oleanolic acid, α-amyrin, and sitosterol) and eluted from the gel in diethyl ether.…”

“…Three fractions, i.e., triterpene acids, combined fraction of free (non-esterified) triterpene alcohols and sterols (referred to as neutral triterpenoids), and triterpenoid esters (extracted from the line of the solvent) were obtained. The fractions containing free triterpene alcohols and sterols were directly analyzed by GC-MS without derivatization, whereas prior to this analysis, fractions containing triterpene acids were methylated with the solution of diazomethane in diethyl ether at 2°C for 24 h, and those containing triterpene esters were subjected to alkaline hydrolysis with 10% NaOH in 80% MeOH at 80°C for 3 h [19][20][21].…”

“…and NIST 2008 Lib. SW (databases version 2010), and where available, by comparison of their retention times and corresponding mass spectra with those of authentic standards [19][20][21][22]. Quantification was based on measuring the area of peaks in the GC-FID chromatograms and performed with the use of external standard method, i.e., calibration curves prepared separately for known concentrations of standards belonging to main triterpenoid classes (α-amyrin for triterpene alcohols, oleanolic acid methyl ester for acids, sitosterol for sterols).…”