Multi-residue determination of 10 selected new psychoactive substances in waste water samples by liquid chromatography–tandem mass spectrometry

New psychoactive substances (NPSs) have become increasingly popular in recent years. The analysis of these substances in influent wastewater (IWW) can be used to track their use in communities. In addition, an evaluation of the amount of NPSs released to the aquatic environment can be performed through the analysis of effluent wastewater (EWW). This study presents the development, validation and application of an analytical methodology, based on solid phase extraction (SPE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS), for the determination of 10 NPSs in IWW and EWW. Synthetic cannabinoids, cathinones, piperazines and pyrrolidophenones are included among the target analytes. To the authors’ knowledge, it is the first time that eight out of these substances (4’-methylpyrrolidinobutyrophenone (MPPP), a-pyrrolidinopentiophenone (a-PVP), 2-[(1S,3R)-3-hydroxycyclohexyl]-5-(2-methyl-2-octanyl) phenol (CP47,497), (1-naphthyl(1-pentyl-1H-indol-3-yl) methanone (JWH-018), (1-butyl-1H-indol-3-yl)(1-naphthyl) methanone (JWH-073), (4-ethyl-1-naphthyl)(1-pentyl-1H-indol-3-yl) methanone (JWH-210), (4-methyl-1-naphthyl) (1-pentyl-1H-indol-3-yl) methanone (JWH-122) and 2-(2-methoxyphenyl)-1-(1-pentyl-1H-indol-3-yl) ethanone (JWH-250)) are investigated in wastewater. The optimized conditions for the analysis of this set of compounds included a SPE clean-up step using a polymeric sorbent and the use of a pentafluorophenyl (PFP) chromatographic column. Despite the broad range of physicochemical properties of the analytes the method allowed acceptable absolute recoveries (40–109%) for all the studied compounds at different levels of concentration. Low method limits of detection (MLODs) were achieved, ranging between 0.3 and 10 ng/L except for BZP and CP47,497 (20 and 23 ng/L, respectively), allowing a reliable and accurate quantification of the analytes. The method was successfully applied to the analysis of IWW and EWW samples from five wastewater treatment plants (WWTPs) located in Santorini Island (a highly touristic resort in Greece). Four out of 10 compounds (a-PVP, CP47,497, JWH-122 and JWH-210) were detected at least in one sample, being the first evidence of their presence in wastewater. CP47,497 was the most ubiquitous and abundant compound, showing concentrations up to 634 ng/L in some case

Highly sensitive determination of 68 psychoactive pharmaceuticals, illicit drugs, and related human metabolites in wastewater by liquid chromatography–tandem mass spectrometry

The present work describes the development and validation of a highly sensitive analytical method for the simultaneous determination of 68 compounds, including illicit drugs (opiates, opioids, cocaine compounds, amphetamines, and hallucinogens), psychiatric drugs (benzodiazepines, barbiturates, anesthetics, antiepileptics, antipsychotics, antidepressants, and sympathomimetics), and selected human metabolites in influent and effluent wastewater (IWWand EWW) by liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS). The method involves a pre-concentration and cleanup step, carried out by solid-phase extraction (SPE) using the adsorbent Strata-XC, followed by the instrumental analysis performed by LC–MS/MS, using a Kinetex pentafluorophenyl (PFP) reversed-phase fused-core column and electrospray ionization (ESI) in both positive and negative modes. A systematic optimization of mobile phases was performed to cope with the wide range of physicochemical properties of the analytes. The PFP column was also compared with two reversed-phase columns: fused-core C18 and XBC18 (with a cross-butyl C18 ligand). SPE optimization and critical aspects associated with the trace level determination of the target compounds (e.g., matrix effects) have been also considered and discussed. Fragmentation patterns for all the classes were proposed. The validated method provides absolute recoveries between 75 and 120 % for most compounds in IWW and EWW. Low method limits of detection were achieved (between 0.04 and 10.0 ng/L for 87 % of the compounds), allowing a reliable and accurate quantification of the analytes at trace level. The method was successfully applied to the analysis of these compounds in five wastewater treatment plants in Santorini, a touristic island of the Aegean Sea, Greece. Thirty-two out of 68 compounds were detected in all IWW samples in the range between 0.6 ng/L (for nordiazepam) and 6,822 ng/L (for carbamazepine) and 22 out of 68 in all EWW samples, with values between 0.4 ng/L (for 9-OH risperidone) and 2,200 ng/L (for carbamazepine). The novel methodology described herein maximizes the information on the environmental analysis of these substances and also provides a first profile of 68 drugs in a Greek touristic are

Hydrophilic Interaction Liquid Chromatography-Electrospray Ionization Mass Spectrometry for Therapeutic Drug Monitoring of Metformin and Rosuvastatin in Human Plasma

In this work a hydrophilic interaction liquid chromatography/positive ion electrospray mass spectrometric assay (HILIC/ESI-MS) has been developed and fully validated for the quantitation of metformin and rosuvastatin in human plasma. Sample preparation involved the use of 100 µL of human plasma, following protein precipitation and filtration. Metformin, rosuvastatin and 4-[2-(propylamino) ethyl] indoline 2 one hydrochloride (internal standard) were separated by using an X-Bridge-HILIC BEH analytical column (150.0 × 2.1 mm i.d., particle size 3.5 µm) with isocratic elution. A mobile phase consisting of 12% (v/v) 15 mM ammonium formate water solution in acetonitrile was used for the separation and pumped at a flow rate of 0.25 mL min−1 . The linear range of the assay was 100 to 5000 ng mL−1 and 2 to 100 ng mL−1 for metformin and rosuvastatin, respectively. The current HILIC-ESI/MS method allows for the accurate and precise quantitation of metformin and rosuvastatin in human plasma with a simple sample preparation and a short a chromatographic run time (less than 15 min). Plasma samples from eight patients were further analysed proving the capability of the proposed method to support a wide range of clinical studies

LB021-MON: The impact of a nutrition workshop on improving adherence to mediterranean diet and nutrition knowledge among adolescent swimmers

Rationale: Adoption of healthy eating by adolescent athletes is imperative for normal growth as well as enhancing sport performance. The study aimed to examine the impact of a healthy eating workshop on adolescent swimmers’ dietary intake, nutrition knowledge and adherence to Mediterranean diet (MD). Methods: Healthy adolescent swimmers aged 13 19 y participated in a half-day educational workshop with emphasis on MD. Adherence to the MD was assessed using the KIDMED index at baseline and 6 weeks post-workshop. Additionally the following were assessed: blood biochemistry, anthropometrics, dietary practices and nutrition knowledge of food sources of macro- and micronutrients and MD definition. Results were analysed by paired t, Wilcoxon and McNemar’s tests as appropriate. Results are presented as mean ± standard deviation or median [interquartile range].Results: A total of 34 subjects participated (67.6% males, 76.5% normal weight and 23.5% overweight). Blood biochemistry was normal both pre- and post-intervention, while BMI was slightly reduced (21.5±2.1 vs 21.4±2.2; p < 0.01).Adherence to the MD was improved (poor: 14.7% vs 2.9%,medium: 64.7% vs 50.0%, good: 20.6% vs 47.1%; KIDMED score:5.00 [3.00] vs 7.00 [2.00]; p < 0.01). Nutrition knowledge also appeared improved (score: 7.00 [3.00] vs 7.00 [2.00]; p = 0.034). Participants reported eating more vegetables, olive oil, pulses and fish and less often in fast food stores (p < 0.05).Conclusion: The KIDMED index is an easy to use dietary assessment tool in this population group. A short educational workshop can improve adherence to MD and nutrition knowledge in adolescent swimmers with possible implications for their health

The impact of nutrition education on nutrition knowledge and adherence to the Mediterranean Diet in adolescent competitive swimmers

Objectives Nutrition education of adolescent competitive swimmers is under-studied although their diet and nutrition knowledge are generally poor. This study aimed to assess the impact of nutrition education on nutrition knowledge and adherence to the Mediterranean Diet (MD) and explore the effect of parental education on the swimmers’ MD adherence. Design A pre–post measurement interventional study was carried out. Methods A half-day nutrition education session was delivered for the swimmers and a separate session for their parents. At baseline and 6-weeks post-workshop, a short nutrition knowledge assessment of food sources of nutrients and the MD composition as well as adherence to the MD using the KIDMED Index were undertaken. The swimmers’ parents also completed the KIDMED Index to evaluate the swimmers’ diet. Results Thirty-four competitive swimmers (age: 15.2 ± 1.5 yr, 23 males) and 22 of their parents participated in the study. There was an improvement in MD adherence with 47% having good adherence post-intervention vs 21% at baseline (p < 0.01) and an increase in the KIDMED Index score (median [interquartile range]: 5.0 [4.0–7.0] vs 7.0 [7.0–9.0]; p < 0.01)). There was also an increase in the swimmers’ nutrition knowledge assessment score (median [IQR]: 7.0 [5.0–8.0] vs 7.0 [6.0–8.0], p < 0.05)), and a trend for a lower score post-intervention in swimmers whose parents did not participate compared to those whose parents participated (6.0 [6.0–7.8] vs 7.0 [7.0–8.0], p = 0.063). Conclusions The intervention improved adherence to the MD and increased nutrition knowledge. The findings support parental participation in nutrition education

Bioanalytical LC–MS Method for the Quantification of Plasma Androgens and Androgen Glucuronides in Breast Cancer

The physiological and pathological development of the breast is strongly affected by the hormonal milieu consisting of steroid hormones. Mass spectrometry (MS) technologies of high sensitivity and specificity enable the quantification of androgens and consequently the characterization of the hormonal status. The aim of this study is the assessment of plasma androgens and androgen glucuronides, in the par excellence hormone-sensitive tissue of the breast, through the application of liquid chromatography–mass spectrometry (LC–MS). A simple and efficient fit-for-purpose method for the simultaneous identification and quantification of dehydroepiandrosterone sulfate (DHEAS), androstenedione (A4), androsterone glucuronide (ADTG) and androstane-3α, 17β-diol-17-glucuronide (3α-diol-17G) in human plasma was developed and validated. The presented method permits omission of derivatization, requires a single solid-phase extraction procedure and the chromatographic separation can be achieved on a single C18 analytical column, for all four analytes. The validated method was successfully applied for the analysis of 191 human plasma samples from postmenopausal women with benign breast disease (BBD), lobular neoplasia (LN), ductal carcinoma in situ and invasive ductal carcinoma (IDC). DHEAS plasma levels exhibited significant differences between LN, IDC and BBD patients (P < 0.05). Additionally, ADTG levels were significantly higher in patients with LN compared with those with BBD (P < 0.05)

Bioanalytical LC-MS Method for the Quantification of Plasma Androgens and Androgen Glucuronides in Breast Cancer

The physiological and pathological development of the breast is strongly affected by the hormonal milieu consisting of steroid hormones. Mass spectrometry (MS) technologies of high sensitivity and specificity enable the quantification of androgens and consequently the characterization of the hormonal status. The aim of this study is the assessment of plasma androgens and androgen glucuronides, in the par excellence hormone-sensitive tissue of the breast, through the application of liquid chromatography-mass spectrometry (LC-MS). A simple and efficient fit-for-purpose method for the simultaneous identification and quantification of dehydroepiandrosterone sulfate (DHEAS), androstenedione (A4), androsterone glucuronide (ADTG) and androstane-3α, 17β-diol-17-glucuronide (3α-diol-17G) in human plasma was developed and validated. The presented method permits omission of derivatization, requires a single solid-phase extraction procedure and the chromatographic separation can be achieved on a single C18 analytical column, for all four analytes. The validated method was successfully applied for the analysis of 191 human plasma samples from postmenopausal women with benign breast disease (BBD), lobular neoplasia (LN), ductal carcinoma in situ and invasive ductal carcinoma (IDC). DHEAS plasma levels exhibited significant differences between LN, IDC and BBD patients (P < 0.05). Additionally, ADTG levels were significantly higher in patients with LN compared with those with BBD (P < 0.05)

Standard addition HPLC method for the determination of a-tocopherol in plasma samples of adolescent swimmers

Vitamin E (a-tocopherol, a-Toc) has been widely used as a powerful antioxidant that protects against oxidation of cellular components. It is used to treat muscular dystrophies, menstrual cycle disorders, risks of pregnancy interruption and abnormalities of gonadal function in men. It is also used frequently from athletes as nutritional supplement for performance enhancing. A simple HPLC method has been validated for the determination of a-Toc in human plasma, using a Nova-Pack analytical column. The chromatographic run time was less than 12 minutes using a mobile phase of Acetonitrile-Methanol 85:15 (v/v), at 0.999 mL/min flow rate while UV/Vis detector was adjusted at 292 nm. The purpose of this study was to evaluate two different approaches for the construction of the calibration curve and further quantify samples from swimmer athletes in order to investigate potential quantification differences. It was shown that the existence of a-Toc as endogenous compounds might affect the actual concentrations and should be considered as an essential parameter during the development of a bioanalytical method for the determination of a-tocopherol in human plasma