An automated platform for multistep synthesis based on a new paradigm for combining flow modules

Automation of organic synthesis has seen rapid progress in the past decade with the development of many platforms for execution, investigation and optimization of chemical reactions and multistep processes. Each of these platforms relies on a different approach to chemical synthesis; including traditional batch chemistry, solid-phase synthesis and flow chemistry, and uses a different dedicated hardware to translate and operate processes. The aim of this thesis was to identify both strengths and limitations of the existing synthesizers and to develop a new platform based on a hybrid approach that takes the best elements from each of them. First, the radial paradigm was established; a new arrangement of reaction modules that allows for their reuse within the same process, uses discrete volumes of solutions and decouples reaction steps so that each reaction in a multistep synthesis can be performed at optimal conditions, minimizing the waste of materials and equipment required (chapter 1, section 1.4). The hardware of the instrument is described thoroughly in chapter 2 with details of each module (broken down into its basic components), explanation of the possible flow pathways, troubleshooting and calibration data. Once the hardware was assembled, the radial synthesizer was validated by performing a series of showcase processes (chapter 3). First, convergent and linear syntheses of the active ingredient rufinamide were chosen to demonstrate the capability of switching between different synthetic routes without the need for physical rearrangement of the instrument (section 3.1). Second, a library of twelve derivatives was generated within a short amount of time and minimum waste of starting materials to show the potential of the radial synthesizer for a possible application in medicinal chemistry (section 3.2). Next, the synthetic and analytical capabilities of the instrument were expanded by integrating a module for photochemistry and flow-NMR spectroscopy via standard flow connectors (section 3.3). Finally, radial synthesis and continuous flow synthesis were compared by preparing three pharmaceutical ingredients (paracetamol, lidocaine, and nifedipine), developing and optimizing each step in the radial synthesizer and performing a scale-up in gram scale in a commercial continuous flow apparatus (section 3.4)

Combining radial and continuous flow synthesis to optimize and scale-up the production of medicines

Current drug production in batch cannot adapt rapidly to market demands, evidenced by recent shortages in many markets globally of essential medicines. Flow chemistry is a valuable tool for on-demand production of active pharmaceutical ingredients (APIs). Here, we reveal a new concept to develop and produce APIs, where an automated synthesizer that works with discrete volumes of solutions is employed at the discovery stage to identify the optimal synthetic route and conditions before a commercially available continuous flow system is used for scale-up. This concept is illustrated by the synthesis of nifedipine and paracetamol, in short supply in Germany during the COVID-19 pandemic, and the local anesthetic lidocaine

Eco-Friendly Engineered Nanomaterials Coupled with Filtering Fine-Mesh Net as a Promising Tool to Remediate Contaminated Freshwater Sludges: An Ecotoxicity Investigation

The use of eco-friendly engineered nanomaterials represents a recent solution for an effective and safe treatment of contaminated dredging sludge. In this study, an eco-designed engineered material based on cross-linked nanocellulose (CNS) was applied for the first time to decontaminate a real matrix from heavy metals (namely Zn, Ni, Cu, and Fe) and other undesired elements (mainly Ba and As) in a lab-scale study, with the aim to design a safe solution for the remediation of contaminated matrices. Contaminated freshwater sludge was treated with CNS coupled with a filtering fine-mesh net, and the obtained waters were tested for acute and sublethal toxicity. In order to check the safety of the proposed treatment system, toxicity tests were conducted by exposing the bacterium Aliivibrio fischeri and the crustacean Heterocypris incongruens, while subtoxicity biomarkers such as lysosomal membrane stability, genetic, and chromosomal damage assessment were performed on the freshwater bivalve Dreissena polymorpha. Dredging sludge was found to be genotoxic, and such genotoxicity was mitigated by the combined use of CNS and a filtering fine-mesh net. Chemical analyses confirmed the results by highlighting the abetment of target contaminants, indicating the present model as a promising tool in freshwater sludge nanoremediation

Cellular Responses Induced by Zinc in Zebra Mussel Haemocytes. Loss of DNA Integrity as a Cellular Mechanism to Evaluate the Suitability of Nanocellulose-Based Materials in Nanoremediation

: Zinc environmental levels are increasing due to human activities, posing a threat to ecosystems and human health. Therefore, new tools able to remediate Zn contamination in freshwater are highly recommended. Specimens of Dreissena polymorpha (zebra mussel) were exposed for 48 h and 7 days to a wide range of ZnCl2 nominal concentrations (1–10–50–100 mg/L), including those environmentally relevant. Cellulose-based nanosponges (CNS) were also tested to assess their safety and suitability for Zn removal from freshwater. Zebra mussels were exposed to 50 mg/L ZnCl2 alone or incubated with 1.25 g/L of CNS (2 h) and then removed by filtration. The effect of Zn decontamination induced by CNS has been verified by the acute toxicity bioassay Microtox®. DNA primary damage was investigated by the Comet assay; micronuclei frequency and nuclear morphological alterations were assessed by Cytome assay in mussels’ haemocytes. The results confirmed the genotoxic effect of ZnCl2 in zebra mussel haemocytes at 48 h and 7-day exposure time. Zinc concentrations were measured in CNS, suggesting that cellulose-based nanosponges were able to remove Zn(II) by reducing its levels in exposure waters and soft tissues of D. polymorpha in agreement with the observed restoration of genetic damage exerted by zinc exposure alon

Analisi della Manutenzione sotto Condizione in "Industrie Cartarie Tronchetti S.p.A."

Negli ultimi anni la competizione tra le imprese è cresciuta esponenzialmente e sono cambiati profondamente i fattori che la caratterizzano, a causa della globalizzazione dei mercati e dell’impiego sempre più intensivo dei capitali; questo ha portato ad un contesto molto turbolento e complesso: un’impresa infatti, per avere successo ed essere efficace sul mercato, deve avere la capacità di interpretare rapidamente i cambiamenti e le richieste dei consumatori e d’altra parte effettuare un miglioramento sistematico dell’utilizzo delle risorse interne per ottenere obiettivi di eccellenza nella gestione, proiettando tutti i settori dell’azienda verso la qualità totale. La ricerca di questi obiettivi deve necessariamente passare attraverso la ridefinizione della funzione della manutenzione e della sua gestione. Nelle aziende la priorità è sempre stata data agli aspetti quantitativi della produzione e la manutenzione, non fornendo alcun prodotto da vendere sul mercato, è stata sempre vista come un costo inevitabile da comprimere il più possibile; essa quindi è stata sempre interpretata come quell’insieme di azioni di carattere correttivo da intraprendere per ripristinare la funzionalità dell’impianto, a seguito di un guasto. Un simile modo di operare ha portato però a risultati assai poco efficienti, visto infatti che i guasti hanno un peso non indifferente sul bilancio dell’impresa in termini di costo di riparazione, perdita di produzione, rischio associato all’evento e, non ultimo, di difetti di qualità associati. Per tutti questi motivi si è sviluppato un nuovo approccio alla manutenzione, che prevede la sua pianificazione ed il progressivo spostamento delle risorse dagli interventi correttivi agli interventi preventivi, predittivi o migliorativi degli impianti, in modo da rendere la manutenzione una funzione efficace ed efficiente e trasformarla in un centro di profitto a tutti gli effetti. Questo passaggio dalla mentalità correttiva a quella proattiva è uno dei punti cardine di strategie operazionali e organizzative recentemente adottate ed apprezzate, come il TPM (Total Productive Maintenance) e la RCM (Reliability Centered Maintenance), che, per la loro completa ed efficace realizzazione, hanno richiesto l’utilizzo di sistemi informatici per la gestione delle attività manutentive, capaci di ridurre i costi associati alla manutenzione: i Computer Managed Maintenance System (CMMS). Tali software sono in grado sia di pianificare e gestire gli interventi di manutenzione e tutto ciò che ne ruota attorno, come il personale addetto, il magazzino ricambi, i fornitori,gli ordini di lavoro, etc., sia di condurre analisi sulla base dati, come l’analisi di criticità. Questa tesi, grazie al continuo contatto con una realtà industriale, si propone di sviluppare conoscenze sul mondo della manutenzione e sulla sua attuale trasformazione nell’azienda e di avere un primo approccio nell’affrontare problematiche di tipo ingegneristico, in particolare in ambito manutentivo. Il lavoro, focalizzandosi proprio su un tipo di software CMMS sviluppato dall’Ingegnere M. Frosolini nell’ambito del progetto Innovazione Distretto Cartario (IN.DI.CA) nella provincia di Lucca e gestito dalla società Lucense, chiamato Coremaint®, ha come obiettivo la sua implementazione, legata alle esigenze dell’azienda, il suo progressivo test e la verifica della sua effettiva utilità. In specifico, dato che in una cartiera il problema della gestione delle attività manutentive risulta di notevole importanza, sia per motivi funzionali che economici, la tesi si propone di effettuare un’analisi dell’attuale organizzazione manutentiva all’interno dell’azienda e di verificare se le metodologie di manutenzione predittiva attualmente adottate portino effettivi benefici. Là dove vi sia un numero elevato di guasti si effettuerà un’analisi al fine di individuarne le cause e alcune metodologie che permettano di monitorare i sintomi scatenanti il gusto così da evitare il fermo dell’attività produttiva (che genera costi di mancata produzione elevati) e il gusto stesso. Il percorso della tesi ha seguito i seguenti successivi passi: 1. inserimento nella realtà aziendale ed analisi della gestione della manutenzione in I.C.T.; 2. formazione, apprendimento e raccolta dati sullo stato attuale del software Coremaint®; 3. analisi delle filosofie manutentive TPM e RCM; 4. applicazione della teoria della scomposizione funzionale alla PM4 nel sito di Pian di Coreglia; 5. caricamento su CoreMaint nel server di I.C.T. della struttura proposta; 6. adeguamento delle strutture esistenti al sistema informatico con l’introduzione di adeguate codifiche dove necessarie; 7. raccolta dati sui guasti che causano la fermata produttiva e dei report tenuti da ISE che effettua la manutenzione predittiva in ICT 8. Individuazione del Sistema/Device più critico attraverso calcolo di indici come FCI e MTBF; 9. Analisi della manutenzione predittiva effettuata sul sistema preso in esame; 10. Analisi dei guasti, individuazione delle cause e indicazione di come applicare la manutenzione sotto condizione per evitare le varie fermate produttive, effettuando un confronto costi /benefici ottenuti dall’applicazione stessa. Il primo e il secondo passo, proprio grazie al contatto diretto con una realtà aziendale, vera utente del software, sono serviti a comprendere le attività svolte in azienda e le varie problematiche ad esse connesse sia dal punto di vista funzionale sia da quello della gestione della manutenzione. Il terzo passo ha previsto un accurato studio delle tecniche tradizionali di manutenzione, delle teorie affidabilistiche più usate nonché di una delle teorie più innovative di gestione della manutenzione (il TPM o Total Productive Maintenance) poiché si è ritenuto che il lavoro non dovesse limitarsi a “fotografare” la situazione esistente ma dovesse contribuire ad un suo miglioramento. Il quarto, il quinto e il sesto passo hanno riguardato l’implementazione del programma Coremaint®, con la collaborazione della software house milanese Omega che lo ha acquistato dal Dipartimento di Produzione di Ingegneria; durante questo lavoro è stata presa in considerazione la macchina PM4 e scomposta funzionalmente analizzando e risolvendo tutti quei problemi relativi alle nuove codifiche fatte. Infine nelle ultime fasi (passo sette, otto, nove e dieci), è stata effettuata un’analisi della gestione della manutenzione in ICT relativa allo stabilimento di Piano di Coreglia e in particolar modo la Paper Machine 4, sulla quale è stata effettuata la scomposizione. E’ stato quindi possibile effettuare un’analisi costi/benefici offerta dall’applicazione di tecnologie della CBM (Condition Based Management) laddove queste non venissero ancora attuate

Nanoparticled Titanium Dioxide to Remediate Crude Oil Exposure. An In Vivo Approach in Dicentrarchus labrax

The contamination of marine water bodies with petroleum hydrocarbons represents a threat to ecosystems and human health. In addition to the surface slick of crude oil, the water-soluble fraction of petroleum is responsible for the induction of severe toxic effects at different cellular and molecular levels. Some petroleum-derived hydrocarbons are classified as carcinogenic and mutagenic contaminants; therefore, the oil spill into the marine environment can have long term consequences to the biota. Therefore, new tools able to remediate crude oil water accommodation fraction pollution in marine water are highly recommended. Nanomaterials were recently proposed in environmental remediation processes. In the present in vivo study, the efficacy of pure anatase titanium nanoparticles (n-TiO2) was tested on Dicentrarchus labrax exposed to the accommodated fraction of crude oil. It was found that n-TiO2 nano-powders themselves were harmless in terms of DNA primary damage, and the capability of pure anatase n-TiO2 to lower the levels of DNA damage induced by a mixture of genotoxic pollutant was revealed. These preliminary results on a laboratory scale are the prerequisite for deepening this new technology for the abatement of the cellular effects related with oil spill pollutants released in marine environments

Eco-Friendly Engineered Nanomaterials Coupled with Filtering Fine-Mesh Net as a Promising Tool to Remediate Contaminated Freshwater Sludges: An Ecotoxicity Investigation

The use of eco-friendly engineered nanomaterials represents a recent solution for an effective and safe treatment of contaminated dredging sludge. In this study, an eco-designed engineered material based on cross-linked nanocellulose (CNS) was applied for the first time to decontaminate a real matrix from heavy metals (namely Zn, Ni, Cu, and Fe) and other undesired elements (mainly Ba and As) in a lab-scale study, with the aim to design a safe solution for the remediation of contaminated matrices. Contaminated freshwater sludge was treated with CNS coupled with a filtering fine-mesh net, and the obtained waters were tested for acute and sublethal toxicity. In order to check the safety of the proposed treatment system, toxicity tests were conducted by exposing the bacterium Aliivibrio fischeri and the crustacean Heterocypris incongruens, while subtoxicity biomarkers such as lysosomal membrane stability, genetic, and chromosomal damage assessment were performed on the freshwater bivalve Dreissena polymorpha. Dredging sludge was found to be genotoxic, and such genotoxicity was mitigated by the combined use of CNS and a filtering fine-mesh net. Chemical analyses confirmed the results by highlighting the abetment of target contaminants, indicating the present model as a promising tool in freshwater sludge nanoremediation

Suitability of Nanoparticles to Face Benzo(a)pyrene-Induced Genetic and Chromosomal Damage in M. galloprovincialis. An In Vitro Approach

Benzo(a)pyrene (B(a)P) is a well-known genotoxic agent, the removal of which from environmental matrices is mandatory, necessitating the application of cleaning strategies that are harmless to human and environmental health. The potential application of nanoparticles (NPs) in the remediation of polluted environments is of increasing interest. Here, specifically designed NPs were selected as being non-genotoxic and able to interact with B(a)P, in order to address the genetic and chromosomal damage it produces. A newly formulated pure anatase nano-titanium (nano-TiO2), a commercial mixture of rutile and anatase, and carbon black-derived hydrophilic NPs (HNP) were applied. Once it had been ascertained that the NPs selected for the work did not induce genotoxicity, marine mussel gill biopsies were exposed in vitro to B(a)P (2 g/mL), alone and in combination with the selected NPs (50 g/mL nano-TiO2, 10 g/mL HNP). DNA primary reversible damage was evaluated by means of the Comet assay. Chromosomal persistent damage was assessed on the basis of micronuclei frequency and nuclear abnormalities by means of the Micronucleus-Cytome assay. Transmission Electron Microscopy (TEM) was performed to investigate the mechanism of action exerted by NPs. Pure Anatase n-TiO2 was found to be the most suitable for our purpose, as it is cyto- and genotoxicity free and able to reduce the genetic and chromosomal damage associated with exposure to B(a)P

An integrated approach for chemical water quality assessment of an urban river stretch through Effect-Based Methods and emerging pollutants analysis with a focus on genotoxicity

The impact of emerging chemical pollutants, on both status and functionality of aquatic ecosystems is worldwide recognized as a relevant issue of concern that should be assessed and managed by researchers, policymakers, and all relevant stakeholders. In Europe, the Reach Regulation has registered more than 100.000 chemical substances daily released in the environment. Furthermore, the effects related to the mixture of substances present in aquatic ecosystems may not be predictable on the basis of chemical analyses alone. This evidence, coupled with the dramatic effects of climate changes on water resources through water scarcity and flooding, makes urgent the application of innovative, fast and reliable monitoring methods. In this context, Effect-Based Methods (EBMs) have been applied in the urban stretch of the Tiber River (Central Italy) with the aim of understanding if detrimental pressures affect aquatic environmental health. In particular, different eco-genotoxicological assays have been used in order to detect genotoxic activity of chemicals present in the river, concurrently characterized by chemical analysis. Teratogenicity and embryo-toxicity have been studied in order to cover additional endpoints. The EBMs have highlighted the presence of diffuse chemical pollution and ecotoxicological effects in the three sampling stations, genotoxicological effects have been also detected through the use of different tests and organisms. The chemical analyses confirmed that in the aquatic ecosystems there is a diffuse presence, even at low concentrations, of emerging contaminants such as pharmaceuticals, not routinely monitored pesticides, personal care products, PFAS. The results of this study can help to identify an appropriate battery of EBMs for future studies and the application of more appropriate measures in order to monitor, mitigate or eliminate chemical contamination and remediate its adverse/detrimental effects on the ecosystem health