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A strategy for test article selection and phytochemical characterization of Echinacea purpurea extract for safety testing.

ABSTRACT: Botanical dietary supplements (BDS) are used around the world for many purported therapeutic properties. The selection of an authentic product and it's phytochemical characterization is critical to generate robust safety data. Because botanicals are complex mixtures with variable quality, identification of a representative product for testing has been challenging. Echinacea is used for its purported immune stimulant properties and was listed as the 2nd top-selling BDS in 2018. However, there are limited safety data for Echinacea. Hence, the National Toxicology Program (NTP) has selected Echinacea for safety testing using rodent models. Here, we describe selection and comprehensive characterization of an Echinacea purpurea root extract to be used in the NTP testing program. Using non-targeted chemical analyses combined with chemometric analysis, a potential unfinished product (i.e., an extract that serves as source material for finished products) of Echinacea purpurea was selected. The product was then authenticated using chemical and DNA techniques and characterized, including the phytochemical composition. Among numerous constituents identified, caftaric acid, chicoric acid, chlorogenic acid and dodeca-2(E),4(E),8(Z),10(E/Z)-tetraenoic acid isobutylamide made up a small fraction of the extract. Based on these analyses, an approach is proposed for test article selection for Echinacea research which can be adapted to other botanicals.

SUBMITTER: Waidyanatha S

PROVIDER: S-EPMC7079738 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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Project description:BACKGROUND: Several Echinacea species have been used as nutraceuticals or botanical drugs for "immunostimulation", but scientific evidence supporting their therapeutic use is still controversial. In this study, a phytocompound mixture extracted from the butanol fraction (BF) of a stem and leaf (S+L) extract of E. purpurea ([BF/S+L/Ep]) containing stringently defined bioactive phytocompounds was obtained using standardized and published procedures. The transcriptomic and proteomic effects of this phytoextract on mouse bone marrow-derived dendritic cells (BMDCs) were analyzed using primary cultures. RESULTS: Treatment of BMDCs with [BF/S+L/Ep] did not significantly influence the phenotypic maturation activity of dendritic cells (DCs). Affymetrix DNA microarray and bioinformatics analyses of genes differentially expressed in DCs treated with [BF/S+L/Ep] for 4 or 12 h revealed that the majority of responsive genes were related to cell adhesion or motility (Cdh10, Itga6, Cdh1, Gja1 and Mmp8), or were chemokines (Cxcl2, Cxcl7) or signaling molecules (Nrxn1, Pkce and Acss1). TRANSPATH database analyses of gene expression and related signaling pathways in treated-DCs predicted the JNK, PP2C-?, AKT, ERK1/2 or MAPKAPK pathways as the putative targets of [BF/S+L/Ep]. In parallel, proteomic analysis showed that the expressions of metabolic-, cytoskeleton- or NF-?B signaling-related proteins were regulated by treatment with [BF/S+L/Ep]. In vitro flow cytometry analysis of chemotaxis-related receptors and in vivo cell trafficking assay further showed that DCs treated with [BF/S+L/Ep] were able to migrate more effectively to peripheral lymph node and spleen tissues than DCs treated as control groups. CONCLUSION: Results from this study suggest that [BF/S+L/Ep] modulates DC mobility and related cellular physiology in the mouse immune system. Moreover, the signaling networks and molecules highlighted here are potential targets for nutritional or clinical application of Echinacea or other candidate medicinal plants.

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A strategy for test article selection and phytochemical characterization of Echinacea purpurea extract for safety testing.

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