Identification of Marker Compounds and In Vitro Toxicity Evaluation of Two Portuguese Asphodelus Leaf Extracts

Abstract

The leaves of Asphodelus bento-rainhae subsp. bento-rainhae, an endemic Portuguese species, and Asphodelus macrocarpus subsp. macrocarpus have been used as food, and traditionally as medicine, for treating ulcers, urinary tract, and inflammatory disorders. The present study aims to establish the phytochemical profile of the main secondary metabolites, together with the antimicrobial, antioxidant and toxicity assessments of both Asphodelus leaf 70% ethanol extracts. Phytochemical screenings were conducted by the TLC and LC-UV/DAD-ESI/MS chromatographic technique, and quantification of the leading chemical classes was performed by spectrophotometric methods. Liquid-liquid partitions of crude extracts were obtained using ethyl ether, ethyl acetate, and water. For in vitro evaluations of antimicrobial activity, the broth microdilution method, and for the antioxidant activity, the FRAP and DPPH methods were used. Genotoxicity and cytotoxicity were assessed by Ames and MTT tests, respectively. Twelve known compounds including neochlorogenic acid, chlorogenic acid, caffeic acid, isoorientin, p-coumaric acid, isovitexin, ferulic acid, luteolin, aloe-emodin, diosmetin, chrysophanol, and β-sitosterol were identified as the main marker compounds, and terpenoids and condensed tannins were found to be the major class of secondary metabolites of both medicinal plants. The ethyl ether fractions demonstrated the highest antibacterial activity against all the Gram-positive microorganisms, (MIC value of 62 to 1000 µg/mL), with aloe-emodin as one of the main marker compounds highly active against Staphylococcus epidermidis (MIC value of 0.8 to 1.6 µg/mL). Ethyl acetate fractions exhibited the highest antioxidant activity (IC50 of 800 to 1200 µg/mL, respectively). No cytotoxicity (up to 1000 µg/mL) or genotoxicity/mutagenicity (up to 5 mg/plate, with/without metabolic activation) were detected. The obtained results contribute to the knowledge of the value and safety of the studied species as herbal medicines.

Quality, Safety and Biological Studies on Campylanthus glaber Aerial Parts

“…CgAE: Campylanthus glaber aqueous extract; CgEE: Campylanthus glaber ethanolic extract; PC (positive control); SD (standard deviation); (1)-2-nitrofluorene; (2)-sodium azide; (3)-tert-butyl hydroperoxide; (4)-9-aminoacridine. * These values are similar to those obtained byMalmir et al (2023) [31] and Malu et al (2022)[32], since this study was performed simultaneously.…”

“…No. HB-8065, Middlesex, UK), according to [31]. Briefly, 48 h after inoculation, the HepG2 culture medium was replaced by fresh medium with CgAE and CgEE (9:1) at final concentrations ranging from 50 to 1000 µg/mL.…”

Multitemporal Land Use and Cover Analysis Coupled with Climatic Change Scenarios to Protect the Endangered Taxon Asphodelus bento-rainhae subsp. bento-rainhae

“…A recent study characterized the substances in the dried root tubers and showed that terpenoids were the main class of secondary metabolites in the extracts [ 24 ]. The leaves of AbR were used as food and in traditional medicine to treat ulcers and urinary and inflammatory disorders [ 25 ], and also as fertilizer and fodder in Portugal [ 26 ]. The ethyl ether fractions demonstrated the highest antibacterial activity against all Gram-positive microorganisms, with aloe-emodin as one of the main marker compounds that are highly active against Staphylococcus epidermidis , and the ethyl acetate fractions exhibited the highest antioxidant activity [ 25 ].…”