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pH induced changes in Raman, UV-Vis absorbance, and fluorescence spectra of dipicolinic acid (DPA)
Umeå University, Faculty of Science and Technology, Department of Physics.
Umeå University, Faculty of Science and Technology, Department of Physics.
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2022 (English)In: Spectrochimica Acta Part A - Molecular and Biomolecular Spectroscopy, ISSN 1386-1425, E-ISSN 1873-3557, Vol. 271, article id 120869Article in journal (Refereed) Published
Abstract [en]

Dipicolinic acid (DPA) is an essential component for the protection of DNA in bacterial endospores and is often used as a biomarker for spore detection. Depending upon the pH of the solution, DPA exists in different ionic forms. Therefore, it is important to understand how these ionic forms influence spectroscopic response. In this work, we characterize Raman and absorption spectra of DPA in a pH range of 2.0–10.5. We show that the ring breathing mode Raman peak of DPA shifts from 1003 cm−1 to 1017 cm−1 and then to 1000 cm−1 as pH increases from 2 to 5. The relative peak intensities related to the different ionic forms of DPA are used to experimentally derive the pKa values (2.3 and 4.8). We observe using UV–vis spectroscopy that the changes in the absorption spectrum of DPA as a function of pH correlate with the changes observed in Raman spectroscopy, and the same pKa values are verified. Lastly, using fluorescence spectroscopy and exciting a DPA solution at between 210–330 nm, we observe a shift in fluorescence emission from 375 nm to 425 nm between pH 2 and pH 6 when exciting at 320 nm. Our work shows that the different spectral responses from the three ionic forms of DPA may have to be taken into account in, e.g., spectral analysis and for detection applications.

Place, publisher, year, edition, pages
Elsevier, 2022. Vol. 271, article id 120869
Keywords [en]
Bacterial spores, DPA, Biomarker, Raman spectra, UV–vis absorption spectra, Fluorescence spectra
National Category
Biophysics Other Physics Topics Inorganic Chemistry
Identifiers
URN: urn:nbn:se:umu:diva-191504DOI: 10.1016/j.saa.2022.120869ISI: 000751812400019PubMedID: 35065519Scopus ID: 2-s2.0-85122995846OAI: oai:DiVA.org:umu-191504DiVA, id: diva2:1629910
Funder
Swedish Research Council, 2019–04016The Kempe Foundations, JCK-1916.2Available from: 2022-01-19 Created: 2022-01-19 Last updated: 2025-09-30Bibliographically approved
In thesis
1. KNOW YOUR ENEMY: Characterizing Pathogenic Biomaterials Using Laser Tweezers
Open this publication in new window or tab >>KNOW YOUR ENEMY: Characterizing Pathogenic Biomaterials Using Laser Tweezers
2022 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Diseases caused by pathogenic agents such as bacteria and viruses result in devastating costs on personal and societal levels. However, it is not just the emergence of new diseases that is problematic. Antibiotic resistance among bacteria makes uncomplicated infections difficult and lethal. Resilient disease-causing spores spread in hospitals, the food industry, and water supplies requiring effective detection and disinfection methods. Further, we face complex neurological diseases where no effective treatment or diagnostic methods exist. Thus, we must increase our fundamental understanding of these diseases to develop effective diagnostic, detection, disinfection, and treatment methods.

Classically, the methods used for detecting and studying the underlying mechanics of pathogenic agents work on a large scale, measuring the average macroscopic behavior and properties of these pathogens. However, just as with humans, the average behavior is not always representative of individual behavior. Therefore, it is also essential to investigate the characteristics of these pathogens on a single cell or particle level. 

This thesis develops and applies optical techniques to characterize pathogenic biomaterial on a single cell or particle level. At the heart of all these studies is our Optical Tweezers (OT) instrument. OT are a tool that allows us to reach into the microscopic world and interact with it. Finally, by combining OT with other experimental techniques, we can chemically characterize biomaterials and develop assays that mimic different biological settings. Using these tools, we investigate bacterial adhesion, disinfection, and detection of pathogenic spores and proteins.

Hopefully, the insights of these studies can lessen the burden on society caused by diseases by helping others develop effective treatment, diagnostic, detection, and disinfection methods in the future. 

Place, publisher, year, edition, pages
Umeå: Umeå universitet, 2022. p. 73
Keywords
Optical Tweezers, Laser Tweezers, Raman Spectroscopy, Bacterial Adhesion, Biophysics, Pili, Bacterial Spores, Endospores, Oocysts, Cryptosporidium, Optics
National Category
Biophysics Atom and Molecular Physics and Optics
Research subject
biology; Physics
Identifiers
urn:nbn:se:umu:diva-192471 (URN)978-91-7855-726-4 (ISBN)978-91-7855-727-1 (ISBN)
Public defence
2022-03-11, NAT.D.410, Naturvetarhuset, Umeå, 09:00 (English)
Opponent
Supervisors
Available from: 2022-02-18 Created: 2022-02-14 Last updated: 2025-02-20Bibliographically approved
2. Spotlight the killer: detecting harmful chemical and biological agents using optical spectroscopy
Open this publication in new window or tab >>Spotlight the killer: detecting harmful chemical and biological agents using optical spectroscopy
2025 (English)Doctoral thesis, comprehensive summary (Other academic)
Alternative title[sv]
Lyset på mördaren : detektion av skadliga kemiska och biologiska ämnen med hjälp av optisk spektroskopi
Abstract [en]

Harmful chemical and biological agents are a significant threat to health and prosperity worldwide. Recent years have seen an increase in wars and conflicts around the globe, raising concerns about the potential deployment of chemical and biological warfare agents. On a less speculative level, harmful chemicals such as narcotic substances cause immense humanitarian and economic damage through overdoses and associated healthcare costs, while microbes such as pathogenic bacteria and parasites cause hospital-acquired infections and food spoilage at a cost of approximately 1 trillion euros every year. To combat the threat of these harmful agents, we must thus develop rapid and effective detection and diagnostic methods for harmful agents, allowing us to effectively deploy specific treatments and preventative measures.

Classically, while there exist numerous methods for the detection of both harmful chemical and biological agents, they often come with limitations that inhibit their effectiveness. These inhibitions often take the form of bulky equipment that is difficult to apply in the field or time-consuming preparation and measurement processes.

In this thesis we will explore an alternative category of assays for detecting and characterizing harmful materials – optical spectroscopy. Optical spectroscopy is a category of material characterization methods that use light to probe a material. While probing the material, we receive a signal characteristic of the molecules, chemical, and biological structure of our material. These optical spectroscopic methods, such as Raman spectroscopy and fluorescence spectroscopy, can be used to characterize a material within the span of minutes or even seconds, making them ideal for detection applications. Furthermore, they can often be made portable or even handheld, making them a great tool for initial field indication of harmful materials, ahead of thorough lab analysis.

I sincerely hope the studies presented herein can serve as a stepping stone to future technologies and detection assays, capable of saving both money and lives. 

Place, publisher, year, edition, pages
Umeå: Umeå University, 2025. p. 72
Keywords
Sensing, Raman spectroscopy, SERS, Fluorescence spectroscopy, CWA, nerve agents, bacterial spores, Cryptosporidium
National Category
Atom and Molecular Physics and Optics
Identifiers
urn:nbn:se:umu:diva-244830 (URN)978-91-8070-780-0 (ISBN)978-91-8070-779-4 (ISBN)
Public defence
2025-10-24, Aula Anatomica, Biologihuset, 907 36, Umeå, Umeå, 13:00 (English)
Opponent
Supervisors
Note

This work was done in collaboration with, and with support from, the Swedish Defece Research Agency (FOI).

Available from: 2025-10-03 Created: 2025-09-30 Last updated: 2025-10-22Bibliographically approved

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Malyshev, DmitryÖberg, RasmusDahlberg, TobiasAndersson, Magnus

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BiophysicsOther Physics TopicsInorganic Chemistry

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