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

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

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

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

Enhancement of Antioxidant Mechanisms and Reduction of Oxidative Stress in Chickens after the Administration of Drinking Water Enriched with Polyphenolic Powder from Olive Mill Waste Waters

The aim of the study was to examine the effects of a polyphenolic powder from olive mill wastewater (OMWW) administered through drinking water, on chickens&apos; redox status. Thus, 75 chickens were divided into three groups. Group A was given just drinking water, while groups B and C were given drinking water containing 20 and 50 μg/ml of polyphenols, respectively, for 45 days. The antioxidant effects of the polyphenolic powder were assessed by measuring oxidative stress biomarkers in blood after 25 and 45 days of treatment. These markers were total antioxidant capacity (TAC), protein carbonyls (CARB), thiobarbituric acid reactive species (TBARS) and superoxide dismutase activity (SOD) in plasma, and glutathione (GSH) and catalase activity in erythrocytes. The results showed that CARB and TBARS were decreased significantly in groups B and C, and SOD decreased in group B compared to that in group A. TAC was increased significantly in group C and GSH was increased in group B, while catalase activity was increased in groups B and C compared to that in group A. In conclusion, this is the first study showing that supplementation of chickens with polyphenols from OMWW through drinking water enhanced their antioxidant mechanisms and reduced oxidative stress-induced damage. © 2017 Aliki Papadopoulou et al

Enhancement of Antioxidant Mechanisms and Reduction of Oxidative Stress in Chickens after the Administration of Drinking Water Enriched with Polyphenolic Powder from Olive Mill Waste Waters

The aim of the study was to examine the effects of a polyphenolic powder from olive mill wastewater (OMWW) administered through drinking water, on chickens' redox status. Thus, 75 chickens were divided into three groups. Group A was given just drinking water, while groups B and C were given drinking water containing 20 and 50 μg/ml of polyphenols, respectively, for 45 days. The antioxidant effects of the polyphenolic powder were assessed by measuring oxidative stress biomarkers in blood after 25 and 45 days of treatment. These markers were total antioxidant capacity (TAC), protein carbonyls (CARB), thiobarbituric acid reactive species (TBARS) and superoxide dismutase activity (SOD) in plasma, and glutathione (GSH) and catalase activity in erythrocytes. The results showed that CARB and TBARS were decreased significantly in groups B and C, and SOD decreased in group B compared to that in group A. TAC was increased significantly in group C and GSH was increased in group B, while catalase activity was increased in groups B and C compared to that in group A. In conclusion, this is the first study showing that supplementation of chickens with polyphenols from OMWW through drinking water enhanced their antioxidant mechanisms and reduced oxidative stress-induced damage. © 2017 Aliki Papadopoulou et al