RESUMEN
Identification and quantification of polyphenols in plant material are of great interest since they make a significant contribution to its total bioactivity. In the present study, an UPLC-Orbitrap-MS/MS approach using the variable data acquisition mode (vDIA) was developed and applied for rapid separation, identification, and quantification of the main polyphenolic compounds in Medicago sativa L. and Trifolium pratense L. sprouts in different germination stages. Based on accurate MS data and fragment ions identification strategy, a total of 29 compounds were identified by comparing their accurate masses, fragment ions, retention times, and literatures. Additionally, a number of 30 compounds were quantified by comparing to the reference standards. Data were statistically analysed. For both plant species, the sprouts of the third germination day are valuable sources of bioactive compounds and could be used in phytotherapy and nutrition. Although Trifolium pratense L. (Red Clover) is considered to be a reference for natural remedies in relieving menopause disorders, alfalfa also showed a high level of biological active compounds with estrogenic activity.
Asunto(s)
Flavonoides/química , Medicago sativa/química , Polifenoles/química , Plantones/química , Trifolium/química , Cromatografía Líquida de Alta Presión , Flavonoides/clasificación , Flavonoides/aislamiento & purificación , Germinación/fisiología , Límite de Detección , Espectrometría de Masas , Medicago sativa/crecimiento & desarrollo , Medicago sativa/metabolismo , Extractos Vegetales/química , Polifenoles/clasificación , Polifenoles/aislamiento & purificación , Estándares de Referencia , Plantones/metabolismo , Factores de Tiempo , Trifolium/crecimiento & desarrollo , Trifolium/metabolismoRESUMEN
The alcoholic extracts from three submerged perennial plants Potamogeton crispus L., P. pusillus L. and P. pectinatus L. were analyzed by gas chromatography-mass spectrometry coupled with solid phase microextraction (SPME-GC/MS) and by High Performance Liquid Chromatography (HPLC) and their volatile fingerprint and polyphenols composition was mutually compared. Twenty-nine chemical compounds were detected and identified in ethanolic and methanolic extracts; the highest abundance (over 5%) in descending order, was detected for 9,9-dimethyl-8,10- dioxapentacyclo (5,3,0(2,5) 0(3,5,)0 (3,6) decane (21.65%), phenol 2,6 bis (1,1 dimethyletyl) 4-1-methylpropil (20.8%), pentadecanoic acid (14.3%), 2-(5-chloro-2-Methoxyphenyl) pyrrole (8.66%), propanedioic (malonic) acid 2-(4-methylphenyl) sulfonyl ethylidene (5.77%), 2 hydroxy-3 tert butyl-5-isopropyl-6 methyl phenyl ketone (5.76%). The highest total polyphenols and flavonoids content was found in the methanolic extract of P. crispus (112.5±0.5 mg tannic acid/g dry extract; 64.2±1.2 mg quercitin/g dry extract). Antioxidant activities (2,2-difenil-1-picrilhidrazil, hydrogen peroxide and reducing power assays) of obtained extracts are comparable with the standard compounds, butylated hydroxytoluene, rutin and ascorbic acid. Antibacterial efficiency of methanolic extracts was notably demonstrated against Gram negative (Escherichia coli, Enterobacter hormaechei) and Gram positive bacteria (Enterococcus casseliflavus). The data reported for the first time for Romanian Potamogeton species, provides extensive support for the chemical investigations of these plants of the aquatic anthropogene ecosystems in order to obtain valuable bioadditives for animal feed and/or pharmaceutical/food industry.