title: Validation of a RP-HPLC-DAD Method for Chamomile (Matricaria recutita) Preparations and Assessment of the Marker, Apigenin-7-glucoside, Safety and Anti-Inflammatory Effect.
abstract: Chamomile is a medicinal plant, which presents several biological effects, especially the anti-inflammatory effect. One of the compounds related to this effect is apigenin, a flavonoid that is mostly found in its glycosylated form, apigenin-7-glucoside (APG), in natural sources. However, the affectivity and safety of this glycoside have not been well explored for topical application. In this context, the aim of this work was to develop and validate a reversed-phase high-performance liquid chromatography (RP-HPLC-DAD) method to quantify APG in chamomile preparations. Additionally, the safety and the anti-inflammatory potential of this flavonoid were verified. The RP-HPLC-DAD method was developed and validated with linearity at 24.0-36.0 μg/mL range (r = 0.9994). Intra- and interday precision (RSD) were 0.27-2.66% and accuracy was 98.27-101.21%. The validated method was applied in the analysis of chamomile flower heads, glycolic extract, and Kamillen cream, supporting the method application in the quality control of chamomile preparations. Furthermore, the APG safety was assessed by MTT cytotoxicity assay and mutagenic protocols and the anti-inflammatory activity was confirmed by a diminished TNF-α production showed by mice macrophages treated with APG following LPS treatment.
authors.affiliation: Laboratório de Pesquisa, Desenvolvimento e Inovação (P, D & I), Apis Flora Industrial e Comercial LTDA, 14020-670 Ribeirão Preto, SP, Brazil.
authors.forename: Felipe Galeti
authors.initials: FG
authors.lastname: Miguel
authors.affiliation: Laboratório de Pesquisa, Desenvolvimento e Inovação (P, D & I), Apis Flora Industrial e Comercial LTDA, 14020-670 Ribeirão Preto, SP, Brazil.
authors.forename: Amanda Henriques
authors.initials: AH
authors.lastname: Cavalheiro
authors.affiliation: Laboratório de Pesquisa, Desenvolvimento e Inovação (P, D & I), Apis Flora Industrial e Comercial LTDA, 14020-670 Ribeirão Preto, SP, Brazil.
authors.forename: Nathália Favaretto
authors.initials: NF
authors.lastname: Spinola
authors.affiliation: Departamento de Genética, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo (FMRP-USP), 14055-370 Ribeirão Preto, SP, Brazil.
authors.forename: Diego Luis
authors.initials: DL
authors.lastname: Ribeiro
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo (FCFRP-USP), 14040-903 Ribeirão Preto, SP, Brazil.
authors.forename: Gustavo Rafael Mazzaron
authors.initials: GR
authors.lastname: Barcelos
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo (FCFRP-USP), 14040-903 Ribeirão Preto, SP, Brazil.
authors.forename: Lusânia Maria Greggi
authors.initials: LM
authors.lastname: Antunes
authors.affiliation: Department of Pharmacology, School of Medicine of Ribeirao Preto, University of Sao Paulo, 14055-370 Ribeirao Preto, SP, Brazil.
authors.forename: Juliana Issa
authors.initials: JI
authors.lastname: Hori
authors.affiliation: Laboratório de Pesquisa, Desenvolvimento e Inovação (P, D & I), Apis Flora Industrial e Comercial LTDA, 14020-670 Ribeirão Preto, SP, Brazil.
authors.forename: Franciane
authors.initials: F
authors.lastname: Marquele-Oliveira
authors.affiliation: Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo (FCFRP-USP), 14040-903 Ribeirão Preto, SP, Brazil.
authors.forename: Bruno Alves
authors.initials: BA
authors.lastname: Rocha
authors.affiliation: Laboratório de Pesquisa, Desenvolvimento e Inovação (P, D & I), Apis Flora Industrial e Comercial LTDA, 14020-670 Ribeirão Preto, SP, Brazil ; Departamento de Ciências Farmacêuticas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo (FCFRP-USP), 14040-903 Ribeirão Preto, SP, Brazil.
authors.forename: Andresa Aparecida
authors.initials: AA
authors.lastname: Berretta