Complementary analytical methods for the phytochemical investigation of ‘Jardin de Granville’, a rose dedicated to cosmetics

Abstract‘Jardin de Granville’ is a new hybrid rose variety dedicated to cosmetic applications. To perform an exhaustive molecular investigation of the non-volatile secondary metabolites in this cultivar, a global approach was developed, combining four chromatographic techniques (HPTLC, HPLC-DAD-ELSD, UHPLC-HRMS and GC–MS). This approach afforded an on-line phytochemical fingerprinting of four plant organs of ‘Jardin de Granville’. Despite the wide diversity of molecular families and the pronounced differences in polarity between the molecules, this analytical strategy enabled an overview of the molecular composition of each sample to be rapidly obtained by HPTLC and HPLC and the molecular content to be correctly identified thanks to coupling with mass spectrometry. Polyphenols were identified in the EtOH/H2O extracts; triterpenes, chlorophyll derivatives and lipids were characterized in the EtOAc extracts, and the fatty acids squalene, α-tocopherol and β-sitosterol were highlighted in the heptane extracts

Phytochemical study of the rose cultivar ‘Jardin de Granville’ : from variety differenciation to molecular specificities

‘Jardin de Granville’ ‘est une variété de rose moderne dédiée à des applications cosmétiques en lien avec ses propriétés intéressantes permettant de lutter contre les mécanismes inflammatoires et oxydants au niveau cutané. L’objectif principal de cette thèse a consisté à établir la cartographie moléculaire de ‘Jardin de Granville’. Pour cela, un procédé standardisé de récolte et d’extraction a été développé afin d’accéder au contenu moléculaire le plus exhaustif possible des différents organes de la plante. Des méthodes complémentaires d’analyse, allant de l’HPTLC, à l’HPLC-DAD-DEDL et jusqu’à l’UHPLC-HRMS, ont été mises en oeuvre pour réaliser les empreintes chromatographiques des extraits et en identifier les principaux constituants. Ces méthodes ont été choisies de plus en plus spécifiques et précises, de façon à apporter une graduation dans le niveau d’informations apportées. Plus de 120 molécules ont pu être caractérisées dans les différents extraits. Le deuxième objectif résidait dans la mise en évidence des marqueurs phytochimiques spécifiques à la variété en comparant ses empreintes moléculaires à celles des deux variétés parents. Deux méthodes de comparaison des profils ont été développées. La première met en jeu des analyses statistiques telles que l’ACP, la CAH et l’ANOVA qui permettent de comparer l’ensemble des extraits. La seconde effectue la soustraction des chromatogrammes d’extrait deux à deux et donne accès à un niveau d’informations plus ciblé. Ces deux approches ont conduit à l’identification de composés différenciant chaque type d’organes ce qui pourra servir d’outils dans la valorisation de certaines parties de la plante. Des marqueurs potentiels plus spécifiques à ‘Jardin de Granville’ ont pu être mis en évidence ce qui démontre la capacité des méthodes développées à différencier le contenu phytochimique de variétés de rose très proches.The modern rose variety ‘Jardin de Granville’, possesses proven activities against skin cell inflammatory and oxidant mechanisms and is devoted to cosmetic applications. The main goal of this study was to establish the molecular fingerprint of the different organs of ‘Jardin de Granville’. In this way, a standardized process for plant harvesting and sample extraction was developed giving access to the most exhaustive molecular fingerprint possible of the different organs. Several complementary analytical methods were implemented through HPTLC, HPLC-DAD-ELSD and UHPLC-HRMS, enabling to achieve the chromatographic fingerprint of the different organs and to identify the main constituents. These methods were selected to have increasing specificity and accuracy to bring progressive information on the molecule structure. Thus, more than 120 compounds were characterized in the different extracts. The second objective consisted in identifying specific phytochemical markers of the variety by comparing its fingerprint to those obtained from its two rose plant parents. In this way, two approaches were developed. The first one involves statistical analysis like PCA, HAC and ANOVA and allows comparing the whole sample chromatograms. The second approach performs extract chromatogram subtractions two by two and gives more detailed information. Both comparative methods led to the identification of the differential compounds existing between the different organ types which could be used to valuate some plant parts in particular. Some ‘Jardin de Granville’ specific markers were highlighted showing the method capacity to distinguish very close rose varieties, by comparing their molecular content

Online acetylcholinesterase inhibition evaluation by high-performance liquid chromatography–mass spectrometry hyphenated with an immobilized enzyme reactor

International audienceA high-performance liquid chromatography-mass spectrometry technique hyphenated on-line with an immobilized enzyme reactor (IMER) was developed by the use of 3 known acetylcholinesterase (AChE) inhibitors (galanthamine, huperzine A and tacrine). This bioanalytical device allows qualitative comparison of the inhibitory strengths of AChE inhibitors. The AChE inhibitory strengths were evaluated and compared by the corresponding acetylcholine peak areas (mass signal) obtained after a chromatographic separation and the elution through the IMER. Only one injection of the analytes is needed to get this comparative analysis. This bioanalytical device was then applied to the extract of a natural plant, Lycoris radiata, which is known to contain AChE inhibitors such as galanthamine and lycoramine. Aside from the demonstration of the inhibitory activity of the two known AChE inhibitors, the AChE inhibitory activity of another compound (dihydro-latifaliumin C) was revealed. This is the first report describing the AChE inhibitory activity of this compound. (C) 2019 Elsevier B.V. All rights reserved

Comparison of Antioxidant Properties of Different Crude Extracts from <i>Citrus natsudaidai</i> Peel

International audienceCitrus natsudaidai is a popular citrus in Japan, but the peel is of limited use and its composition is not well documented. The purposes of this study were to obtain health-beneficial compounds from C. natsudaidai peel for food use by simple methods and to investigate the antioxidant capacities of extracts of different polarities without overestimating the effects. C. natsudaidai peel was extracted with water-ethanol and hexane-ethanol solutions to obtain five different soluble fractions. The highest total phenolic content (TPC) was obtained from the water-soluble fraction (WSF), and the lowest content was observed in the hexane soluble fraction (HSF). The WSF also showed the highest antioxidant activities, and the lowest activities were found in HSF. A positive correlation was detected between TPC and antioxidant activities. The polar extract of C. natsudaidai peel has potential as a natural additive or ingredient in foods, cosmetics and pharmaceutical products with antioxidant activity

Molecular Fingerprint Comparison of Closely Related Rose Varieties based on UHPLC‐HRMS Analysis and Chemometrics

Abstract Introduction The “Jardin de Granville” modern rose variety not only combines the morphological properties of its two parental cultivars, but also possesses better agronomic characteristics (abundant blooms, strong growth and vitality, high resistance to common rose diseases). In addition, it shows remarkable biological properties such as a high ability to decrease inflammatory and oxidative stress on skin cells. That is why Parfums Christian Dior selected this rose variety to be an active ingredient in luxury cosmetics. Objectives To identify the characteristic molecular signature of “Jardin de Granville” compared with its parents “Annapurna” and “John Clare”, by the mean of a non‐targeted metabolomic comparison. Material and Methods Wood, flower and leaf hydro‐alcoholic extracts were analysed by UHPLC‐ESI‐HRMS. The fingerprints were then submitted to unsupervised multivariate analyses involving principal component analysis (PCA) and hierarchical ascendant classification (HAC). Analysis of variance (ANOVA) was finally performed to highlight the significant differences in each group of organs. Results The extracts were composed of phenolic compounds such as hydrolysable and condensed tannins and flavonol derivatives. Three groups of extracts were clustered as a function of the variety. The compounds overexpressed in “Jardin de Granville” variety were highlighted thanks to ANOVA test. Flower was the most discriminative organ with 15 overexpressed molecules. Auto MS/MS analyses led to their tentative identifications. Conclusion The non‐targeted metabolomic approach revealed the importance of tannins to discriminate close rose varieties. The overexpressed hydrolysable tannins characteristic of “Jardin de Granville” can be responsible for the antioxidant and anti‐inflammatory properties of the rose cosmetic ingredients. Copyright © 2016 John Wiley & Sons, Ltd

Severity of manchineel fruit (Hippomane mancinella) poisoning: A retrospective case series of 97 patients from French Poison Control Centers

International audienceIn this retrospective series of 97 cases of manchineel fruit ingestion reported to French Poison Control Centers between 2009 and 2017, we investigated cases of poisoning due to manchineel fruit (from the Hippomane mancinella tree). This fruit is known to be responsible for oropharyngeal and gastrointestinal tract lesions and possibly hypotension and bradycardia (previously attributed to the presence of physostigmine). The most commonly observed clinical signs were oropharyngeal pain, abdominal pain, diarrhea and oropharyngeal irritation. No major gastrointestinal tract lesions were observed in the five cases in which upper gastrointestinal (GI) endoscopy was performed. One case of laryngeal edema and one case of bradycardia were observed, but analysis of the harvested fruits did not confirm the presence of physostigmine. Ingestion of manchineel fruit can cause mild abdominal pain and digestive irritation, requiring medical attention. Rarely, when several fruits have been ingested, severe oropharyngeal injury or hemodynamic disorders may require otorhinolaryngological consultation or cardiac monitoring for several hours, respectively

Clerodane furanoditerpenoids as the probable cause of toxic hepatitis induced by Tinospora crispa

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Complementary Strategies to Unlock Biosynthesis Gene Clusters Encoding Secondary Metabolites in the Filamentous Fungus Podospora anserina

International audienceThe coprophilous ascomycete Podospora anserina is known to have a high potential to synthesize a wide array of secondary metabolites (SMs). However, to date, the characterization of SMs in this species, as in other filamentous fungal species, is far less than expected by the functional prediction through genome mining, likely due to the inactivity of most SMs biosynthesis gene clusters (BGCs) under standard conditions. In this work, our main objective was to compare the global strategies usually used to deregulate SM gene clusters in P. anserina, including the variation of culture conditions and the modification of the chromatin state either by genetic manipulation or by chemical treatment, and to show the complementarity of the approaches between them. In this way, we showed that the metabolomics-driven comparative analysis unveils the unexpected diversity of metabolic changes in P. anserina and that the integrated strategies have a mutual complementary effect on the expression of the fungal metabolome. Then, our results demonstrate that metabolite production is significantly influenced by varied cultivation states and epigenetic modifications. We believe that the strategy described in this study will facilitate the discovery of fungal metabolites of interest and will improve the ability to prioritize the production of specific fungal SMs with an optimized treatment

Complementary Strategies to Unlock Biosynthesis Gene Clusters Encoding Secondary Metabolites in the Filamentous Fungus <i>Podospora anserina</i>

The coprophilous ascomycete Podospora anserina is known to have a high potential to synthesize a wide array of secondary metabolites (SMs). However, to date, the characterization of SMs in this species, as in other filamentous fungal species, is far less than expected by the functional prediction through genome mining, likely due to the inactivity of most SMs biosynthesis gene clusters (BGCs) under standard conditions. In this work, our main objective was to compare the global strategies usually used to deregulate SM gene clusters in P. anserina, including the variation of culture conditions and the modification of the chromatin state either by genetic manipulation or by chemical treatment, and to show the complementarity of the approaches between them. In this way, we showed that the metabolomics-driven comparative analysis unveils the unexpected diversity of metabolic changes in P. anserina and that the integrated strategies have a mutual complementary effect on the expression of the fungal metabolome. Then, our results demonstrate that metabolite production is significantly influenced by varied cultivation states and epigenetic modifications. We believe that the strategy described in this study will facilitate the discovery of fungal metabolites of interest and will improve the ability to prioritize the production of specific fungal SMs with an optimized treatment

Plant-microbe features of Dendrobium fimbriatum (Orchidaceae) fungal community

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