Phytochemical Screening and Acanthamoebic Activity of Shoots from in Vitro Cultures and in Vivo Plants of Eryngium alpinum L.—The Endangered and Protected Species

Abstract

Genetically uniform shoots of Eryngium alpinum L. cultured in vitro were subjected to the qualitative analysis applying the UPLC-HESI-HRMS technique. In vitro cultures give the opportunity to perform the phytochemical studies on the protected species without harvesting the plant material from the natural environment. The phytochemical screening of the crude methanolic extracts of shoots, both from in vitro cultures and in vivo plants, revealed the presence of phenolic acids, coumarins, flavonoids, triterpenoid saponins, amino acids, or dipeptides. Active compounds detected are known to have medicinal importance, and for this reason, the present study represents a preliminary investigation of the extracts against pathogenic and opportunistic amoeba. Among the extracts tested, the extract of shoots from in vitro cultures exhibited remarkable amoebicidal action against trophozoites. On the second day of treatment, the extract at the concentrations of 5 mg/mL, 2.5 mg/mL, and 0.5 mg/mL showed the highest antiamoebicidal effect: the inhibition of trophozoites reached 81.14%, 66.38%, and 54.99%, respectively. To our best knowledge, the present report is the first to show the phytochemical screening and to discuss the antiamoebic activity of Eryngium alpinum L. shoots, both from in vitro cultures and in vivo plants.

Chemical Composition of Essential Oils from Rare and Endangered Species—Eryngium maritimum L. and E. alpinum L.

“…[20]. The phytochemical investigations of E. alpinum revealed the presence of secondary metabolites-triterpenoid saponins [21], phenolic acids mainly caffeic acid, rosmarinic acid and its derivative R-(+)-3'-O-β-d-glucopyranosyl rosmarinic acid, chlorogenic, iso-and neochlorogenic acids, and 3,4-dihydroxyphenylacetic acid [21][22][23], flavonoids-kaempferol, quercetin, and isoquercetin [21,23,24], and the essential oil [25]. The analysis of the essential oil of the E. alpinum aboveground part performed by Dunkić et al [25], showed the presence of oxygenated sesquiterpenes, sesquiterpene hydrocarbons, oxygenated monoterpenes, and monoterpene hydrocarbons.…”

Two Ecdysteroids Isolated from Micropropagated Lychnis flos-cuculi and the Biological Activity of Plant Material

“…The activity of L. flos-cuculi, compared to the amoebicidal activity of extracts from other plant species, is fairly high. For example, the effective concentration of an Eryngium alpinum shoot culture extract was 0.5 mg/mL, displaying a similarly high potency of an in vitro-derived material extract rich in triterpenoid saponins [ 39 ]. However, the comparison of IC 50 values for extracts of plants especially effective against Acanthamoeba , including Solidago virgaurea flowers (0.01 mg/mL), Solidago graminifolia flowers (0.05 mg/mL), Pueraria lobata root (0.01 mg/mL), and Rubus chamaemorus leaves (0.05 mg/mL) [ 40 ], shows that only the 80% methanolic fraction of the L. flos-cuculi root extract exerts a similarly strong activity, with IC 50 at 0.06 mg/mL.…”

Effect of Elicitation with (+)-Usnic Acid on Accumulation of Phenolic Acids and Flavonoids in Agitated Microshoots of Eryngium alpinum L.

“…Phenolic acids (chlorogenic acid, rosmarinic acid, and its derivative R-(+)-3′ O - β -D-glucopyranosyl rosmarinic acid [ 7 ], as well as caftaric acid, neochlorogenic acid, isochlorogenic acid, 3,4-dihydroxyphenylacetic acid, and caffeic acid [ 8 , 9 ]) have previously been found in this species. Moreover, the group of hydroxycinnamic acid derivates: coumaroylcholine, caffeoylcholine, feruloylquinic acid, dicaffeoylquinic acid, and ferulic acid, and methyl rosmarinate, as well as benzoic acid derivates: glucosyringic acid, trimethoxybenzoic acid, vanillic acid, hydroxyphenyllactic acid, dimethoxybenzaldehyde, glucovanillin, have also been found [ 10 ]. Additionally, flavonoids (quercetin and kaempferol [ 11 ], isoquercetin and quercitrin [ 8 , 9 ], luteolin, luteolin 7- O -glucoside, kaempferol-3- O -rhamnoside, kaempferol-3- O -rutinoside, quercetin-3- O -glucoside, quercetin-3- O -galactoside, quercetin-3- O -rutinoside [ 10 ]) and coumarins (umbelliferone, scopoletin, 4-methylumbelliferyl glucuronide, fraxidin, isofraxidin, scopoline, esculin, dihydroxycoumarin) have been found in this species [ 10 ].…”

“…Moreover, the group of hydroxycinnamic acid derivates: coumaroylcholine, caffeoylcholine, feruloylquinic acid, dicaffeoylquinic acid, and ferulic acid, and methyl rosmarinate, as well as benzoic acid derivates: glucosyringic acid, trimethoxybenzoic acid, vanillic acid, hydroxyphenyllactic acid, dimethoxybenzaldehyde, glucovanillin, have also been found [ 10 ]. Additionally, flavonoids (quercetin and kaempferol [ 11 ], isoquercetin and quercitrin [ 8 , 9 ], luteolin, luteolin 7- O -glucoside, kaempferol-3- O -rhamnoside, kaempferol-3- O -rutinoside, quercetin-3- O -glucoside, quercetin-3- O -galactoside, quercetin-3- O -rutinoside [ 10 ]) and coumarins (umbelliferone, scopoletin, 4-methylumbelliferyl glucuronide, fraxidin, isofraxidin, scopoline, esculin, dihydroxycoumarin) have been found in this species [ 10 ]. The metabolites detected were triterpenoid saponins (3- O – β -D-glucopyranosyl-(1→2)- β -D-glucuronopyranosyl-22- O -angeloyl-R1-barrigenol, 3- O - β -D-glucopyranosyl-(1→2)- β -D-glucuronopyranosyl-22- O -angeloyl-A1-barrigenol) and eryngioside J [ 10 ].…”