Workflow for the identification of biotransformation products of amine-containing psychotropic drugs in the aquatic environment

Pharmaceuticals are continuously discarded Pharmaceuticals are continuously discarded into the aquatic system through wastewater treatment plants (WWTPs). The microbial degradation of these organic micropollutants and formation of transformation products (TPs) under aerobic conditions is the fundamental process for their elimination. It is of paramount importance to understand the microbial metabolic pathways so as to obtain knowledge of how fast micropollutants degraded and to assess the exposure to their potential TPs as they can be more polar and consequently environmentally persistent. In this study, batch reactors seeded with activated sludge from the WWTP of Athens were set up to assess biotic, abiotic and sorption losses of selective psychotropic drugs, containing amine moieties. Biodegradation and transformation products were identified using liquid chromatography quadrupole-time-offlight mass spectrometry (LC-QToF-MS). A workflow for target, suspect and non-target screening was developed. Data treatment was performed by using metabolite tools accompanying Bruker’s maxis impact ESI-QToF-MS and the structure elucidation of the candidate transformation products was based on accurate mass and isotopic pattern measurements by HRMS and tentative interpretation of MS/MS spectra. Finally four biotransformation products were identified for both lidocaine and ephedrine. Despite the structure similarities, different degradation constants were calculated for each compound

Well-defined homopolypeptides, copolypeptides, and hybrids of Poly(l-proline)

l-Proline is the only, out of 20 essential, amino acid that contains a cyclized substituted α-amino group (is formally an imino acid), which restricts its conformational shape. The synthesis of well-defined homo- and copolymers of l-proline has been plagued either by the low purity of the monomer or the inability of most initiating species to polymerize the corresponding N-carboxy anhydride (NCA) because they require a hydrogen on the 3-N position of the five-member ring of the NCA, which is missing. Herein, highly pure l-proline NCA was synthesized by using the Boc-protected, rather than the free amino acid. The protection of the amine group as well as the efficient purification method utilized resulted in the synthesis of highly pure l-proline NCA. The high purity of the monomer and the use of an amino initiator, which does not require the presence of the 3-N hydrogen, led for the first time to well-defined poly(l-proline) (PLP) homopolymers, poly(ethylene oxide)-b-poly(l-proline), and poly(l-proline)-b-poly(ethylene oxide)-b-poly(l-proline) hybrids, along with poly(γ-benzyl-l-glutamate)-b-poly(l-proline) and poly(Boc-l-lysine)-b-poly(l-proline) copolypeptides. The combined characterization (NMR, FTIR, and MS) that results for the l-proline NCA revealed its high purity. In addition, all synthesized polymers exhibit high molecular and compositional homogeneity

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

Non-target approach for the determination of novel micropollutants in wastewater using liquid chromatography quadrupole-time of flight mass spectrometry (LC-QTOF-MS)

Wastewaters contain a very large list of micropollutants and transformation products of environmental concern. All these (mostly) synthetic organic chemicals enter the wastewater treatment plants (WWTP) with influents and due to incomplete or zero removal are released in the aquatic environment. Thus, the study of the fate of the emerging pollutants and their transformation products in WWTPs is of paramount environmental importance and can also provide valuable information related to consumption trends. Target screening procedures are limited to a small fraction of these substances, due to the inability to obtain standards for all that substances and the ignorance of the existence of many of them. Recent advances in high resolution mass spectrometry (HRMS) have opened up new windows of opportunity in the field of complex samples analysis. Suspect screening, with suspected substances based on prior information but with no reference standard, is a powerful tool which allows a large increment in the number of compounds to be evaluated. However, in most cases many of the peaks showing greater intensity not correspond to substances included in the target and suspect screening lists. These substances are potentially relevant, due to their high concentration, and their identification is environmentally important. Nevertheless, full identification of unknown compounds is often difficult and there is no guarantee of a successful outcome. The aim of the present work is the development and application of a workflow for the tentative identification of relevant unknown substances (not detected in the previously applied target and suspect methods) using liquid chromatography quadrupole-time-of-flight mass spectrometry (LC–QToF-MS)

Identification of unknowns in real wastewater through the application of a LC-QTOF-MS based workflow

Wastewater contains a high number of organic micropollutants and transformation products of environmental concern. Recent approaches, combining methodologies based on target and suspect screening (for suspected substances based on prior information but with no reference standard) are important for the comprehensive characterization of environmental samples. Nevertheless, samples still contain many chromatographic peaks which do not correspond to substances included in target and suspect screening lists. These substances may be potentially relevant (e.g. due to their concentration or potential effects) and thus the identification of selected non-targets is important. However, full identification of unknown compounds is often difficult and there is no guarantee of a successful outcome. The aim of this work is to show some specific examples on the identification of unknown compounds in real wastewater (collected from the WWTP of Athens). Identifications were conducted using a developed integrated workflow based on LC–QToF-MS to detect formerly unknown organic contaminants in wastewater