Periodic Reporting for period 1 - SPOTplasma (Flexible Tube Micro Plasma (FμTP) for Dried Blood Spot (DBS) analysis)
Reporting period: 2021-05-01 to 2023-04-30
SPOTplasma aims at developing innovative solutions for the growing field of direct biofluid spot (DBFS) analysis on paper. The main goal was to enhance the efficiency, speed, cost-effectiveness, and accessibility of this analytical process. DBFS analysis holds significant importance in various topics, including newborn screening (NBS), pharmacokinetics, and doping testing. Particularly in NBS, the adoption of techniques that expedite the process (first-tier) can help mitigate potential health issues in neonates and reduce the analytical expenses, thereby facilitating access to NBS analysis in resource-constrained settings.
The conventional methods utilized for DBFS sample analysis are characterized by lengthy extraction procedures, arduous derivatization reactions, substantial solvent usage, and reliance on sophisticated instrumentation for separation and detection. However, this project has introduced innovative tools for the direct analysis of DBFS utilizing ambient ionization (AI) mass spectrometry (MS). Innovative methodologies have been developed and refined, harnessing the potential of a flexible Tube Micro Plasma (FμTP), cost-effective desorption methods like handheld IR-Laser, and the integration of paper spray ionization with MS. These techniques hold great promise as first-tier methods for NBS analysis or directly analyze DBFS. Through the identification and optimization of the factors influencing ion source performance, and analytical methodologies, this project has not only displayed significant advancements but also nurtured the development of new technologies that will contribute to the enhancement of DBFS analysis.
The conventional methods utilized for DBFS sample analysis are characterized by lengthy extraction procedures, arduous derivatization reactions, substantial solvent usage, and reliance on sophisticated instrumentation for separation and detection. However, this project has introduced innovative tools for the direct analysis of DBFS utilizing ambient ionization (AI) mass spectrometry (MS). Innovative methodologies have been developed and refined, harnessing the potential of a flexible Tube Micro Plasma (FμTP), cost-effective desorption methods like handheld IR-Laser, and the integration of paper spray ionization with MS. These techniques hold great promise as first-tier methods for NBS analysis or directly analyze DBFS. Through the identification and optimization of the factors influencing ion source performance, and analytical methodologies, this project has not only displayed significant advancements but also nurtured the development of new technologies that will contribute to the enhancement of DBFS analysis.
SPOTplasma has been persistent in its pursuit of developing innovative tools to transform the analysis of DBFS through the exploration of novel and affordable methodologies based in AI-MS. The project was focused on uncovering and harnessing the latent potential of the FμTP as an ionization source for DBFS. The distinctive nature of this ion source renders it both alluring and formidable. To better understand the ion source, an evaluation was conducted to ascertain its ionization capacity comparing it to analogous plasma-based ion sources. Remarkably, the FμTP exhibited a comparable internal energy distribution to its counterparts. However, the miniaturization character of the ion source proved to be a double-edged sword, as the low flow rates presented challenges in the direct analysis of DBFS.
A variety of approaches was explored, all while staying true to the core principle of SPOTplasma: rapid and easily accessible methodologies. One example was the exploration of a low-cost desorption tool, a handheld IR laser, followed by subsequent ionization utilizing the FμTP. When the laser was utilized, rapid pulses of heat were precisely administered to the paper, mitigating the risk of paper combustion or long waiting times associated with conventional heat sources. This method not only demonstrated the efficacy of carefully selected extraction solvents in conjunction with optimized laser and FμTP conditions, but also facilitated the direct analysis of drugs in biofluids such as saliva, blood, plasma, and other samples deposited on paper, including beverages and tap water.
Indeed, the paper serves not only as a substrate for DBFS collection but can also serve as the substrate for ionization. Paper spray-MS has emerged as an ideal technique for the direct analysis of biological samples, including urine, tissue, and blood, crucial for disease detection and diagnosis. This is largely attributed to the paper substrate's minimal sample consumption, stability, affordability, and ease of disposal. Furthermore, the paper spray-MS methodology requires a few experimental prerequisites, encompassing voltage, solvent selection, and detection protocols. To this end, a refined approach involving thick film reaction on paper derivatization was investigated to facilitate the swift diagnosis of amino acid-based metabolic disorders, an area of utmost importance in NBS. A pioneering first-tier method was proposed, enabling NBS analysis to be completed in less than 10 minutes.
The achievements of SPOTplasma have highlighted the importance of innovative research approaches and the pivotal role of AI-MS in the implementation of cutting-edge tools within the realm of clinical chemistry. The project has yielded significant outcomes, with select results already published in peer-reviewed journals and further endeavors currently underway. These remarkable findings have been presented at international conferences, with dedicated efforts directed toward disseminating knowledge on plasma-based ionization, NBS, and MS, thus extending the project's reach to the wider public.
A variety of approaches was explored, all while staying true to the core principle of SPOTplasma: rapid and easily accessible methodologies. One example was the exploration of a low-cost desorption tool, a handheld IR laser, followed by subsequent ionization utilizing the FμTP. When the laser was utilized, rapid pulses of heat were precisely administered to the paper, mitigating the risk of paper combustion or long waiting times associated with conventional heat sources. This method not only demonstrated the efficacy of carefully selected extraction solvents in conjunction with optimized laser and FμTP conditions, but also facilitated the direct analysis of drugs in biofluids such as saliva, blood, plasma, and other samples deposited on paper, including beverages and tap water.
Indeed, the paper serves not only as a substrate for DBFS collection but can also serve as the substrate for ionization. Paper spray-MS has emerged as an ideal technique for the direct analysis of biological samples, including urine, tissue, and blood, crucial for disease detection and diagnosis. This is largely attributed to the paper substrate's minimal sample consumption, stability, affordability, and ease of disposal. Furthermore, the paper spray-MS methodology requires a few experimental prerequisites, encompassing voltage, solvent selection, and detection protocols. To this end, a refined approach involving thick film reaction on paper derivatization was investigated to facilitate the swift diagnosis of amino acid-based metabolic disorders, an area of utmost importance in NBS. A pioneering first-tier method was proposed, enabling NBS analysis to be completed in less than 10 minutes.
The achievements of SPOTplasma have highlighted the importance of innovative research approaches and the pivotal role of AI-MS in the implementation of cutting-edge tools within the realm of clinical chemistry. The project has yielded significant outcomes, with select results already published in peer-reviewed journals and further endeavors currently underway. These remarkable findings have been presented at international conferences, with dedicated efforts directed toward disseminating knowledge on plasma-based ionization, NBS, and MS, thus extending the project's reach to the wider public.
SPOTplasma demonstrated the potential of the proof-of-concept research paving the way for innovative solutions to enhance the analysis of DBFS. The utilization of cost-effective techniques has not only facilitated rapid diagnosis and detection but also increased affordability, enabling a broader spectrum of users to benefit from these advancements.
The findings of the project serve as a solid foundation for further research and development. Future efforts will be focused on refining the techniques and methods employed, exploring high-throughput approaches, and seeking more cost-effective couplings. Building upon these achievements, a new research line has been initiated within the hosting group, demonstrating a commitment to the research launched during the project.
In public research, the pursuit of new tools for health promotion and early detection holds immense importance. Recognizing this, the European Commission has included in the Horizon Europe 2021-2027 program a dedicated research Cluster focused on Health (Cluster 2). The outcomes and discoveries of the SPOTplasma project align with two key impacts identified in the program: Impact 4, which aims to ensure access to innovative, sustainable, and high-quality healthcare, and Impact 5, which strives to unlock the full potential of new tools, technologies, and digital solutions for a healthier society. The project's novel tools contribute significantly to the achievement of these program objectives, highlighting their value in addressing critical societal needs.
The findings of the project serve as a solid foundation for further research and development. Future efforts will be focused on refining the techniques and methods employed, exploring high-throughput approaches, and seeking more cost-effective couplings. Building upon these achievements, a new research line has been initiated within the hosting group, demonstrating a commitment to the research launched during the project.
In public research, the pursuit of new tools for health promotion and early detection holds immense importance. Recognizing this, the European Commission has included in the Horizon Europe 2021-2027 program a dedicated research Cluster focused on Health (Cluster 2). The outcomes and discoveries of the SPOTplasma project align with two key impacts identified in the program: Impact 4, which aims to ensure access to innovative, sustainable, and high-quality healthcare, and Impact 5, which strives to unlock the full potential of new tools, technologies, and digital solutions for a healthier society. The project's novel tools contribute significantly to the achievement of these program objectives, highlighting their value in addressing critical societal needs.