Bryophyllum pinnatum markers: CPC isolation, simultaneous quantification by a validated UPLC-DAD method and biological evaluations.

Autor: Morais Fernandes J; Grupo de Pesquisa em Produtos Naturais Bioativos, Departamento de Farmácia, Universidade Federal do Rio Grande do Norte, 59.012-570 Natal, Brazil., Ortiz S; Equipe Produits Naturels, Analyse, Synthèse, UMR CNRS 8038, Faculté de Pharmacie, Equipe Produits Naturels, Analyse, Synthèse, UMR CNRS 8038, Faculté de Pharmacie, Université de Paris, 75006, Paris, France., Padilha M Tavares R; Grupo de Pesquisa em Produtos Naturais Bioativos, Departamento de Farmácia, Universidade Federal do Rio Grande do Norte, 59.012-570 Natal, Brazil., Mandova T; Equipe Produits Naturels, Analyse, Synthèse, UMR CNRS 8038, Faculté de Pharmacie, Equipe Produits Naturels, Analyse, Synthèse, UMR CNRS 8038, Faculté de Pharmacie, Université de Paris, 75006, Paris, France; Gilson Purification SAS, Saint-Avé, France., Rodrigues D Araújo E; Grupo de Pesquisa em Produtos Naturais Bioativos, Departamento de Farmácia, Universidade Federal do Rio Grande do Norte, 59.012-570 Natal, Brazil., L Andrade AW; Grupo de Pesquisa em Produtos Naturais Bioativos, Departamento de Farmácia, Universidade Federal do Rio Grande do Norte, 59.012-570 Natal, Brazil., Michel S; Equipe Produits Naturels, Analyse, Synthèse, UMR CNRS 8038, Faculté de Pharmacie, Equipe Produits Naturels, Analyse, Synthèse, UMR CNRS 8038, Faculté de Pharmacie, Université de Paris, 75006, Paris, France., Grougnet R; Equipe Produits Naturels, Analyse, Synthèse, UMR CNRS 8038, Faculté de Pharmacie, Equipe Produits Naturels, Analyse, Synthèse, UMR CNRS 8038, Faculté de Pharmacie, Université de Paris, 75006, Paris, France. Electronic address: raphael.grougnet@parisdescartes.fr., Zucolotto SM; Grupo de Pesquisa em Produtos Naturais Bioativos, Departamento de Farmácia, Universidade Federal do Rio Grande do Norte, 59.012-570 Natal, Brazil. Electronic address: silvanazucolotto@ufrnet.br.
Jazyk: angličtina
Zdroj: Journal of pharmaceutical and biomedical analysis [J Pharm Biomed Anal] 2021 Jan 30; Vol. 193, pp. 113682. Date of Electronic Publication: 2020 Oct 12.
DOI: 10.1016/j.jpba.2020.113682
Abstrakt: Bryophyllum pinnatum (Lam.) Oken (Crassulaceae) is widely used as leaf juice or extracts in traditional medicine all over tropical areas, especially in Brazil, to relieve inflammation-associated symptoms. Flavonol glycosides with unusual sugar moiety are among the major metabolites. Nevertheless, there are not enough quality control studies that can contribute to authentication of B. pinnatum and determination of their markers. As it is also used as medicinal plant in several countries, it is necessary to provide data related to safety, efficacy and quality. In this context, this work aims to isolate the major flavonoids from B. pinnatum hydroethanolic extract, to validate a method to quantify the content of chemical markers and to evaluate their xanthine oxidase inhibition and antioxidant activity. The extract was submitted to centrifugal partition chromatography (CPC). The solvents system CyHex-EtOAc-EtOH-H 2 O, 0.5:9:3:5.5, v/v/v/v was selected by shake-flask method. Four flavonoids (quercetin 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (1), kaempferol 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (2), quercetin 3-O-α-L-rhamnopyranoside (3) and kaempferol 3-O-α-L-rhamnopyranoside (4)) were isolated in a single and fast CPC run and their structures were confirmed by NMR analysis. An UPLC-DAD quantification method was established for the first time with validation of required parameters, according to RDC 166/2017. The calibration curves were linear with correlation coefficient ranging from 0.9996 to 0.9997 while the values of LOD (0.0077-1.984 ng.mL -1 ), LOQ (0.0263-6.012 ng.mL -1 ), recovery (≥ 80.7 %) and inter-day (%RSD ≤ 3.581) and intra-day precision (%RSD ≤ 2.628) were satisfactory. Quantitative analysis of these compounds showed that the proportion of 1, 2 and 3 were 2.43, 0.25 and 0.33 % (24.3 mg.g -1 , 0.25 mg.g -1 and 0.33 mg.g -1 of extract), respectively. Moreover, in vitro xanthine oxidase (XO), DPPH and ABTS inhibition were evaluated for the extract and the major flavonoids. Compounds 2 (168 μM) and 3 (124 μM) moderately inhibited XO, while compounds 1 and 3 displayed average radical scavenging activity. In conclusion, our results suggest the flavonoid 1 as a specific marker which may be used for quality control of B. pinnatum hydroethanolic leaves extract.
Competing Interests: Declaration of Competing Interest The authors report no declarations of interest.
(Copyright © 2020 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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