Accurate quantification of cellular metabolites is essential for understanding the biochemical processes underlying various diseases. This study investigates the application of Surface-Enhanced Raman Spectroscopy (SERS) for the semiquantitative analysis of adenine, used here as a model cellular metabolite. A SERS-based approach was selected due to its high sensitivity and specificity in detecting low concentrations of chemical species. The method involved the preparation of active substrates in the form of thin films composed of silver and gold nanoparticles, followed by the deposition of adenine solutions at different concentrations. The results demonstrate that this approach enables the detection and quantification of low adenine concentrations with high specificity. The semiquantitative nature of the method was validated by comparing SERS signal intensities with known adenine concentrations, revealing a strong correlation. In conclusion, this technique provides a powerful analytical tool for biochemical research and holds significant potential for extension to the analysis of a broader range of metabolites. Future work will focus on optimizing the method for in vitro applications and exploring its use in the investigation of additional cellular components.
La quantificazione accurata dei metaboliti cellulari è fondamentale per comprendere i vari processi biochimici alla base, per esempio di varie malattie. Questo studio esplora l'applicazione della Spettroscopia Raman Amplificata da Superficie (SERS) per l'analisi semiquantitativa di Adenina, come esempio di metabolita cellulare. Abbiamo scelto un approccio basato sul SERS per la sua alta sensibilità e specificità nel rilevare basse concentrazioni di specie chimiche. Il metodo ha comportato la preparazione di substrati attivi, nella forma di film sottili di nanoparticelle di argento e oro e successiva applicazione con soluzioni di Adenina a varie concentrazioni. I risultati dimostrano che l’approccio utilizzato può rilevare e quantificare basse concentrazioni di Adenina con alta specificità. La natura semiquantitativa del metodo è stata convalidata confrontando i segnali SERS con concentrazioni conosciute di Adenina, mostrando una forte correlazione. In conclusione, questa tecnica offre uno strumento potente per la ricerca biochimica e ha il potenziale per essere estesa all'analisi di una gamma più ampia di metaboliti. I lavori futuri si concentreranno sul perfezionamento della tecnica per applicazioni in vitro ed esploreranno il suo utilizzo per ulteriori componenti cellulari.
Metodi semiquantitavi per metaboliti cellulari mediante tecnica SERS
BORDIGNON, MASSIMILIANO
2024/2025
Abstract
Accurate quantification of cellular metabolites is essential for understanding the biochemical processes underlying various diseases. This study investigates the application of Surface-Enhanced Raman Spectroscopy (SERS) for the semiquantitative analysis of adenine, used here as a model cellular metabolite. A SERS-based approach was selected due to its high sensitivity and specificity in detecting low concentrations of chemical species. The method involved the preparation of active substrates in the form of thin films composed of silver and gold nanoparticles, followed by the deposition of adenine solutions at different concentrations. The results demonstrate that this approach enables the detection and quantification of low adenine concentrations with high specificity. The semiquantitative nature of the method was validated by comparing SERS signal intensities with known adenine concentrations, revealing a strong correlation. In conclusion, this technique provides a powerful analytical tool for biochemical research and holds significant potential for extension to the analysis of a broader range of metabolites. Future work will focus on optimizing the method for in vitro applications and exploring its use in the investigation of additional cellular components.| File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.12608/100380