Phytochemical analysis and antioxidant potential of the phytonutrient-rich decoction of Cichorium spinosum and C. intybus.

The Cretan diet as the basis of the Mediterranean diet, has provided traditional remedies for the general well-being through the long-established consumption of cooked wild greens and vegetables. The intake of water decoctions of Cichorium spinosum and Cichorium intybus in the context of the daily dietary regime in Greece, has been long associated with “liver detoxifying” properties. In the current study, we performed an in depth investigation of the water decoctions traditionally prepared from C. spinosum and C. intybus, through qualitative UHPLC-HRMS profiling and direct quantification of cichoric and caftaric acid, as major antioxidant components of the decoction. In addition, we developed a one-step Countercurrent Chromatography method for the isolation of the two phenolic acids, along with a sulfoconjugate sesquiterpene lactone present only in the Cretan C. spinosum. All water decoctions were found not cytotoxic in human fibroblasts, whereas they all significantly reduce the intracellular reactive oxygen species, in consistency with the major presence of strong antioxidant compounds such as cichoric acid. This work demonstrates that the intake of decoction in doses suggested by the Greek traditional use is comparable to the ingestion of a phytomedical preparation of antioxidants. These results contribute to our current knowledge on the beneficial health effect of the Cretan diet

Caractérisation de la contamination des systèmes aquatiques par les psychotropes : développement d'une méthode de référence incluant une démarche multifactorielle de hiérarchisation

Pharmaceuticals and their residues belong to the so-called “emerging” pollutants for which no current regulation is applied regarding their presence in the environment. Monitoring the contamination of aquatic resources by these kinds of micropollutants is a necessity in order to anticipate potential effects regarding human health and ecosystems. In this context, data quality (traceability and comparability) is compulsory so as to take better decisions regarding policies to be driven. Psychotropic compounds, which are widely consumed, display a great concern in that field.This thesis aimed at characterizing occurrence of psychotropic compounds in the environment and putting forward metrological tools insuring accuracy, traceability, comparability as well as representativity of the measurement results.To that purpose, a SPE-LC-MS² multi-residue method has been developed for a selection of psychotropic compounds and other commonly consumed drugs. Method limits of quantification were in the range of ng.L-1 which enabled to characterize their occurrence in several surface waters and treated wastewaters. Concurrently, usage of integrative samplers was evaluated and applied on the field in different locations. Occurrence levels of 68 selected compounds were established by implementation of this complementary strategy in aquatic systems. Furthermore, results from measurement campaigns were used to rank studied compounds into a short list of interest. Finally, a reference method using isotopic dilution and mass spectrometry (DI-MS) was developed for the characterization of 24 psychotropic compounds in surface waters. Expanded uncertainties (k=2) inferior to 10% were obtained. Last be not least, implementing this method to environmental analysis emphasized the importance of sampling related uncertainties regarding to global measurement uncertainties.Les résidus de médicaments font partie des polluants dits « émergents » pour lesquels il n’existe actuellement pas de réglementation quant à leur présence dans l’environnement. Afin d’anticiper un risque éventuel pour l’Homme et les milieux aquatiques il est, entre autres, nécessaire de réaliser un état des lieux de la contamination des systèmes aquatiques par ce type de micropolluants. Dans ce contexte, disposer de données de mesures de qualité (traçables et comparables) constitue un prérequis indispensable pour décider au mieux des politiques à conduire dans ce domaine. L’étude des psychotropes présente un enjeu important d’autant plus qu’il s’agit de molécules abondamment consommées.Les travaux conduits lors de cette thèse avaient deux objectifs principaux. Le premier était de caractériser l’occurrence environnementale de composés psychotropes. Le second objectif était de proposer des outils métrologiques permettant d’assurer l’exactitude, la traçabilité, la comparabilité mais également la représentativité des résultats de mesure.Dans un premier temps, une méthode multi-résidus par SPE-LC-MS² a été développée pour une sélection de composés psychotropes et d’autres médicaments de consommation courante. Les limites de quantification de la méthode étaient de l’ordre du ng.L-1. Cela a permis de caractériser leur occurrence dans différents cours d’eau et rejets de station d’épuration en France. En parallèle, une approche par échantillonnage intégratif a été développée et déployée dans différents sites. Cette stratégie complémentaire a permis d’établir les niveaux d’occurrence des 68 psychotropes et traceurs sélectionnés et de conduire une démarche de hiérarchisation dans le but d’établir une liste restreinte de composés d’intérêt. Enfin, une méthode de référence associant la dilution isotopique à la spectrométrie de masse (DI-MS) a été développée pour l’analyse de 24 psychotropes dans les eaux de surface. Elle se caractérise par des incertitudes élargies (k=2) inférieures à 10%. La mise en oeuvre de cette méthode dans des études environnementales a permis de mettre en évidence l’importance de la contribution de l’incertitude liée à l’échantillonnage par rapport à l’incertitude totale de mesure

Characterization of the contamination of aquatic systems by psychotropic compounds : development of a reference method including a multifactorial selection approach

Les résidus de médicaments font partie des polluants dits « émergents » pour lesquels il n’existe actuellement pas de réglementation quant à leur présence dans l’environnement. Afin d’anticiper un risque éventuel pour l’Homme et les milieux aquatiques il est, entre autres, nécessaire de réaliser un état des lieux de la contamination des systèmes aquatiques par ce type de micropolluants. Dans ce contexte, disposer de données de mesures de qualité (traçables et comparables) constitue un prérequis indispensable pour décider au mieux des politiques à conduire dans ce domaine. L’étude des psychotropes présente un enjeu important d’autant plus qu’il s’agit de molécules abondamment consommées.Les travaux conduits lors de cette thèse avaient deux objectifs principaux. Le premier était de caractériser l’occurrence environnementale de composés psychotropes. Le second objectif était de proposer des outils métrologiques permettant d’assurer l’exactitude, la traçabilité, la comparabilité mais également la représentativité des résultats de mesure.Dans un premier temps, une méthode multi-résidus par SPE-LC-MS² a été développée pour une sélection de composés psychotropes et d’autres médicaments de consommation courante. Les limites de quantification de la méthode étaient de l’ordre du ng.L-1. Cela a permis de caractériser leur occurrence dans différents cours d’eau et rejets de station d’épuration en France. En parallèle, une approche par échantillonnage intégratif a été développée et déployée dans différents sites. Cette stratégie complémentaire a permis d’établir les niveaux d’occurrence des 68 psychotropes et traceurs sélectionnés et de conduire une démarche de hiérarchisation dans le but d’établir une liste restreinte de composés d’intérêt. Enfin, une méthode de référence associant la dilution isotopique à la spectrométrie de masse (DI-MS) a été développée pour l’analyse de 24 psychotropes dans les eaux de surface. Elle se caractérise par des incertitudes élargies (k=2) inférieures à 10%. La mise en oeuvre de cette méthode dans des études environnementales a permis de mettre en évidence l’importance de la contribution de l’incertitude liée à l’échantillonnage par rapport à l’incertitude totale de mesure.Pharmaceuticals and their residues belong to the so-called “emerging” pollutants for which no current regulation is applied regarding their presence in the environment. Monitoring the contamination of aquatic resources by these kinds of micropollutants is a necessity in order to anticipate potential effects regarding human health and ecosystems. In this context, data quality (traceability and comparability) is compulsory so as to take better decisions regarding policies to be driven. Psychotropic compounds, which are widely consumed, display a great concern in that field.This thesis aimed at characterizing occurrence of psychotropic compounds in the environment and putting forward metrological tools insuring accuracy, traceability, comparability as well as representativity of the measurement results.To that purpose, a SPE-LC-MS² multi-residue method has been developed for a selection of psychotropic compounds and other commonly consumed drugs. Method limits of quantification were in the range of ng.L-1 which enabled to characterize their occurrence in several surface waters and treated wastewaters. Concurrently, usage of integrative samplers was evaluated and applied on the field in different locations. Occurrence levels of 68 selected compounds were established by implementation of this complementary strategy in aquatic systems. Furthermore, results from measurement campaigns were used to rank studied compounds into a short list of interest. Finally, a reference method using isotopic dilution and mass spectrometry (DI-MS) was developed for the characterization of 24 psychotropic compounds in surface waters. Expanded uncertainties (k=2) inferior to 10% were obtained. Last be not least, implementing this method to environmental analysis emphasized the importance of sampling related uncertainties regarding to global measurement uncertainties

Validation and uncertainties evaluation of an isotope dilution-SPE-LC–MS/MS for the quantification of drug residues in surface waters

International audienceThe present work describes the development and validation of a reference method conducted at the French National Institute of Metrology (LNE) for the quantitative determination of psychoactive compounds in the dissolved fraction of surface waters. More specifically an isotope dilution-SPE-LC-MS/MS based method has been implemented for the characterization of a broad range of analytes belonging to different classes of psychotropic drugs such as benzodiazepines, antidepressants, stimulants, opiates and opioids, anticonvulsants, anti-dementia drugs, analgesics as well as the anti-inflammatory drug diclofenac in the low ng L(-1) range of concentration. Full validation of the method was performed following procedures described by the French standard NF T90-210. Limits of quantification between 0.14 and 3.54 ng L(-1) were obtained. Method recoveries from 71 to 123% were observed with standard deviation below 10% in intermediate precision conditions. Accuracy was determined for every compound: measurement errors were between -4 and +1% and standard deviations in intermediate precision conditions were included within a 1-9% interval. Finally, measurement uncertainties were evaluated following the Guide to the expression of uncertainty in measurement (GUM). Expanded uncertainties (k=2) ranged from 2% for carbamazepine, EDDP (2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine) and venlafaxine to 17% for diazepam. The validated method was implemented to Seine river surface waters demonstrating its fitness for purpose. All compounds were detected and 22 out of 25 analytes were quantified. More specifically, measured concentration ranged from 0.39 ng L(-1) for MDMA (3,4-methylene-dioxy-N-methylamphetamine) to 182 ng L(-1) for gabapentine

Development of a multi-residue method for scrutinizing psychotropic compounds in natural waters

International audienceThe present work describes a multi-residue SPE-UPLC–MS/MS method aiming at the characterization of 68 compounds in natural waters, including parent compounds as well as their major metabolites and glucuronide conjugates. Development was conducted toward the quantitative determination of a broad range of analytes belonging to different class of psychotropic drugs such as benzodiazepines, antidepressants, stimulants, opiates and opioids, anticonvulsants, anti-dementia drugs, analgesic and anti-inflammatory drugs (as anthropic indicators) in the low ng L−1 range of concentration. Satisfactory extraction recoveries >70% were obtained for the majority of analytes (49 out of 68) allowing low limits of quantification. LOQ ranged between 0.1 and 17.8 ng L−1 and were lower than 5 ng L−1 for 94% of investigated analytes. Furthermore, addition of 25 isotopic labeled standards allowed to ensure reliability of the optimized method. Quantification errors were typically below 15% with relative standard variations <10% in intermediate precision conditions. Finally, the developed method was implemented in natural waters; sampling campaigns were conducted in the Seine River as a demonstration of the applicability and adequation of the method for its purpose. As a result, 48 out of 68 analytes were identified or quantified; some of them like memantine, rivastigmine, zolpidem 4-phenyl-carboxylic acid, zolpidem 6-carboxylic acid for one of the first time in surface waters. Among investigated psychotropic compounds and metabolites, tramadol, codeine, oxazepam, venlafaxine, O-desmethylvenlafaxine, gabapentin, carbamazepine and 10,11-dihydro-10,11-dihydroxycarbamazepine were found to be the most abundant

Bioavailability of Terpenes and Postprandial Effect on Human Antioxidant Potential. An Open-Label Study in Healthy Subjects

Scope: To assess bioavailability of terpenes in human plasma and their effect on oxidative stress biomarkers. Methods and results: In this open-label and single arm postprandial trial, seventeen healthy male volunteers (20–40 years old) follow a low-phytochemical diet for 5 days. Next, after overnight fasting, volunteers consume Mastiha powder (a natural resin rich in terpenes) dispersed in water. Blood samples are collected on time points 0 h (before ingestion) and 0.5, 1, 2, 4, 6, and 24 h (post-ingestion). Ultra-high-pressure liquid chromatography high-resolution MS (UHPLC-HRMS/MS) is applied for high throughput analysis of plasma. Serum resistance to oxidation and oxidized LDL (oxLDL) levels are measured. UHPLC-HRMS/MS analysis shows that major terpenes are bioavailable since 0.5 h after administration, reaching a peak between 2 h and 4 h. Serum resistance to oxidation, expressed as difference of tLAG (time point-0 h), starts to increase from 0.5 h. This increase reaches statistical significance at 4 h (402.3 ± 65.0 s), peaks at 6 h (524.6 ± 62.9 s), and remains statistically significant until 24 h (424.2 ± 48.0 s). oxLDL levels, expressed as %change from 0 h, are reduced significantly from time point-1 h until time point-6 h. Conclusion: Results demonstrate the terpene bioavailability pattern after oral administration of Mastiha. Terpenes are potential mediators of antioxidant defense in vivo. © 2017 WILEY-VCH Verlag GmbH &amp; Co. KGaA, Weinhei

UHPLC/HR-ESI-MS/MS Profiling of Phenolics from Tunisian Lycium arabicum Boiss. Antioxidant and Anti-lipase Activities’ Evaluation

This study was performed in the aim to evaluate nine different extracts from Tunisian Lycium arabicum for their total phenolic and total flavonoid contents, phytochemical analyses as well as their antioxidant and anti-lipase activities. The in vitro antioxidant property was investigated using three complementary methods (DPPH, ferric reducing antioxidant power (FRAP), and β-carotene-linoleic acid bleaching assays) while anti-lipase activity was evaluated using 4-methylumbelliferyl oleate method. From all of the tested extracts the most potent found to be the polar MeOH extracts especially those of stems and leaves. In order to investigate the chemical composition of these extracts and possible correlation of their constituents with the observed activities, an UHPLC/HR-ESI-MS/MS analysis was performed. Several compounds belonging to different chemical classes were tentatively identified such as rutin and kampferol rutinoside, the major constituents of the leaves, and N-caffeoyltyramine, lyciumide A, N-dihydrocaffeoyltyramine as well as fatty acids: trihydroxyoctadecadienoic acid and hydroxyoctadecadienoic acid isomers were detected abundantly in the stems. These results showed that the MeOH extracts of stems and leaves of L. arabicum can be considered as a potential source of biological active compounds. © 2017 Wiley-VHCA AG, Zurich, Switzerlan

Chemical Composition, Antibacterial Activity using Micro-broth Dilution Method and Antioxidant Activity of Essential Oil and Water Extract from Aerial Part of Tunisian Thymus algeriensis Boiss. &amp; Reut.

In this study, we investigated the chemical composition, antioxidant and antibacterial activities of essential oil and aqueous extract from Thymus algeriensis Boiss. & Reut growing in Tunisia. GC/MS analysis of essential oil from the aerial part of the plant led to identifying 54 constituents representing 96.87 % of the total oil composition. Monoterpenes represented the major components of the essential oil. On the other hand, the aqueous extract and the essential oil were screened for their antioxidant and antibacterial activities. The latter was assessed against Gram-negative and Gram-positive bacterial strains using the broth dilution micro method for the determination of antibacterial activity. The antioxidant activity was evaluated using the DPPH scavenging activity and Ferric Reducing/Antioxidant Power (FRAP) assays. The essential oil was found to be more active in antibacterial screening (MIC = 0.54 μg/mL). Whereas the aqueous extract exhibited the most potent antioxidant activity with (IC50 = 0.04 μg/mL). UHPLC ESI(-)-HRMS/MS analysis of the aqueous extract allowed tentative identification of its constituents and showed that it is rich in phenolic compounds. Luteolin-glucuronide was found to be the most abundant compound followed by Vicenin-2 and Apigenin-diglucuronide.This work was supported by the Ministry of Higher Education, Scientific Research and Technologies, Tunisia, allowed to the Laboratory of Organic Chemistry, Natural Substances Section (LR17/ ES08), Faculty of Sciences of Sfax, University of Sfax. The authors S. Fontanay and R.E. Duval are grateful to the Ministry of Higher Education, Research and Innovation, France, CNRS and Région Grand-Est which financed part of this work. The authors are thankful to Professor Leandros A Skaltsounis, Division of Pharmacognosy and Natural Products Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens for precious facilities in LCESI-HRMS/MS analysis

Chemical Composition, Antibacterial Activity using Micro-broth Dilution Method and Antioxidant Activity of Essential Oil and Water Extract from Aerial Part of Tunisian Thymus algeriensis Boiss. &amp; Reut.

In this study, we investigated the chemical composition, antioxidant and antibacterial activities of essential oil and aqueous extract from Thymus algeriensis Boiss. &amp;amp; Reut growing in Tunisia. GC/MS analysis of essential oil from the aerial part of the plant led to identifying 54 constituents representing 96.87 % of the total oil composition. Monoterpenes represented the major components of the essential oil. On the other hand, the aqueous extract and the essential oil were screened for their antioxidant and antibacterial activities. The latter was assessed against Gram-negative and Gram-positive bacterial strains using the broth dilution micro method for the determination of antibacterial activity. The antioxidant activity was evaluated using the DPPH scavenging activity and Ferric Reducing/Antioxidant Power (FRAP) assays. The essential oil was found to be more active in antibacterial screening (MIC = 0.54 μg/mL). Whereas the aqueous extract exhibited the most potent antioxidant activity with (IC50 = 0.04 μg/mL). UHPLC ESI(-)-HRMS/MS analysis of the aqueous extract allowed tentative identification of its constituents and showed that it is rich in phenolic compounds. Luteolin-glucuronide was found to be the most abundant compound followed by Vicenin-2 and Apigenin-diglucuronide. © 2022 Har Krishan Bhalla &amp; Sons