{"id":65133,"date":"2026-06-22T14:03:38","date_gmt":"2026-06-22T17:03:38","guid":{"rendered":"https:\/\/cnpem.br\/?p=65133"},"modified":"2026-06-22T14:05:24","modified_gmt":"2026-06-22T17:05:24","slug":"protocol-combines-x-rays-and-electrochemistry-to-study-hydrogen-production","status":"publish","type":"post","link":"https:\/\/cnpem.br\/en\/protocol-combines-x-rays-and-electrochemistry-to-study-hydrogen-production\/","title":{"rendered":"Protocol combines X-rays and electrochemistry to study hydrogen production"},"content":{"rendered":"

[vc_row][vc_column][vc_column_text css=””]<\/p>\n

Experimental protocol details challenges and solutions for the study of biocatalysts in beamlines with nanometric beams, such as Carna\u00faba, at Sirius<\/p><\/blockquote>\n

A\u00a0<\/span>work published in the<\/span>\u00a0scientific<\/span>\u00a0journal Nature Protocols<\/span><\/a>\u00a0by researchers from<\/span>\u00a0the\u00a0S\u00e3o Carlos\u202f<\/span>Institute of<\/span>\u202fChemistry<\/span>\u00a0at\u00a0<\/span>the University of S\u00e3o Paulo (IQSC-USP) in collaboration with the Brazilian Center for Research in Energy and Materials (CNPEM) presents a detailed guide for the study of biocatalysts under operational conditions, combining X-ray absorption spectroscopy with electrochemical techniques. <\/span>The article brings together years of experimental development and describes how to study proteins that participate in reactions relevant to various applications, including energy production.<\/span><\/p>\n

<\/p>\n

Biocatalysts\u00a0and\u00a0green\u00a0energy<\/h3>\n

The central motivation of this work is the use of biocatalysts to enable cleaner and more efficient chemical processes. Unlike conventional synthetic catalysts, biocatalysts are three-dimensional, operate under mild conditions, and have high selectivity. This reduces not only the environmental impact, but also operational costs and risks associated with chemical processes. <\/span>\u00a0<\/span><\/p>\n

Furthermore,\u00a0<\/span>several of the reactions of interest already occur in biological systems, which allows<\/span>\u00a0researchers<\/span>, instead of recreating complex processes artificially, to take advantage of and adapt mechanisms that have been refined by nature throughout evolution.<\/span>\u00a0<\/span>Another critical point is the use of\u00a0metals.<\/span>\u00a0<\/span>Industrial catalysts often rely on elements such as platinum or iridium, which are highly efficient but extremely expensive and scarce.<\/span>\u00a0<\/span>Biocatalysts, on the other hand, can offer a more accessible alternative, especially when derived from abundant materials, including plants and fungi.<\/span>\u00a0<\/span>\u00a0<\/span><\/p>\n

According\u00a0to<\/span>\u00a0Itamar Neckel, CNPEM researcher at the Carna<\/span>\u00fa<\/span>ba\u00a0beamline and co-author of the article, this is one of the main obstacles to the application of these technologies:<\/span>\u00a0\u201c<\/span>It is possible to use platinum and iridium to break down the water molecule and therefore to produce hydrogen, but these elements are expensive<\/span>.\u201d<\/span>\u00a0<\/span><\/p>\n

In the context of hydrogen production, these characteristics are particularly relevant.<\/span>\u00a0<\/span>Reactions such as water oxidation, which are fundamental to the generation of this fuel, can be mediated by enzymes with specific metal sites, usually of more abundant and cheaper metals, such as copper, for example.<\/span>\u00a0<\/span>Studying these systems is an essential step towards developing more sustainable energy technologies.<\/span>\u00a0<\/span>\u00a0<\/span><\/p>\n

XA-SEC: combining X-rays and electrochemistry<\/h3>\n

To investigate these processes in detail, the study uses an approach known as<\/span>\u00a0spectroelectrochemistry,\u00a0<\/span>which is the simultaneous performance of electrochemistry experiments with X-ray absorption, or XA-SEC.<\/span>\u00a0<\/span>In practice, this means combining two techniques: applying an electrical stimulus to the material and, at the same time, measuring how the metal inside the enzyme absorbs X-rays.<\/span><\/p>\n

\"\"

The electrochemical cell installed at the Tarum\u00e3 station, on the Carna\u00faba beamline, in Sirius. The combination of electrochemistry and X-rays makes it possible to monitor, in real time, structural and electronic changes in biocatalysts during reactions of interest for the production of sustainable energy.<\/p><\/div>\n

When an electrical potential is applied, the oxidation state of the metal present in the catalyst can change.<\/span>\u00a0<\/span>These changes directly affect the energy at which the material absorbs X-rays.<\/span>\u00a0<\/span>By\u00a0monitoring\u00a0this behavior, researchers\u00a0are able to\u00a0monitor, in real time, how the catalyst responds to the reaction conditions, as well as understand the step-by-step process of the reaction.<\/span>\u00a0<\/span>\u00a0<\/span><\/p>\n

Each element has a characteristic\u00a0<\/span>\u201c<\/span>absorption edge<\/span>\u201d<\/span>, an energy range where a significant variation in photon absorption occurs.<\/span>\u00a0<\/span>Changes in the oxidation state shift this edge, allowing the identification of electronic transformations in the material<\/span>.\u00a0<\/span>\u00a0<\/span><\/p>\n

The technique’s distinguishing feature lies precisely in its simultaneity: while different potentials are applied, absorption spectra are also measured.<\/span>\u00a0<\/span>This allows for a direct correlation between electrochemical behavior and changes in the electronic structure of the catalyst, an important window for\u00a0observing\u00a0ongoing reactions.<\/span>\u00a0<\/span>\u00a0<\/span><\/p>\n

Experimental\u00a0challenges<\/h3>\n

Despite the wealth of information, carrying out this type of experiment is far from trivial.<\/span>\u00a0<\/span>One of the first challenges is assembling the electrochemical cell, which requires extremely clean and carefully prepared electrodes.<\/span>\u00a0<\/span><\/p>\n

\"\"

Electrochemical cell used in XAS-SEC experiments. Inside the device, biocatalysts immobilized on an electrode are subjected to different electrical potentials while their electronic properties are investigated by X-ray absorption spectroscopy.<\/p><\/div>\n

The enzyme of interest needs to be immobilized on the working electrode, usually from a solution that is deposited on the surface.<\/span>\u00a0<\/span>Next, a polymer is added to stabilize the material and allow the passage of cations, preventing the enzyme from dissolving in the aqueous medium or undergoing degradation.<\/span>\u00a0<\/span>\u00a0<\/span><\/p>\n

Another critical point is the concentration of the material.<\/span>\u00a0<\/span>Very small\u00a0amounts make it difficult to detect the X-ray signal emitted by the sample.<\/span>\u00a0<\/span>However,\u00a0a very high\u00a0concentration implies an excessively thick layer of enzymes, causing poor electronic connection due to high electrical resistance.<\/span>\u00a0<\/span>Finding this balance requires multiple tests and preparation of\u00a0<\/span>several<\/span>\u00a0samples until a good region to perform measurements is found.<\/span>\u00a0<\/span>\u00a0<\/span><\/p>\n

Besides\u00a0this<\/span>, there is\u00a0the\u00a0problem of radiation damage.<\/span>\u00a0<\/span>The intensity of the X-ray beam in high-resolution beamlines, such as Carna<\/span>\u00fa<\/span>ba, can alter the material itself that is being studied, leading to<\/span>\u00a0its<\/span>\u00a0degradation.<\/span>\u00a0<\/span>Therefore, it is necessary to carry out preliminary tests to ensure that the observed changes are in fact a result of the experiment and not the action of the beam.<\/span>\u00a0<\/span>Add<\/span>ing\u00a0<\/span>to\u00a0th<\/span>is<\/span>\u00a0are challenges such as precise alignment of the sample in the experimental station and control of the photon flow, which needs to be\u00a0optimized\u00a0to balance signal quality and material preservation.<\/span>\u00a0<\/span>\u00a0<\/span><\/p>\n

The importance of experimental protocols<\/h3>\n

Given the many variables and challenges faced by researchers worldwide, the work developed by these researchers offers a clear systematization of these complex steps in a clear and reproducible protocol.<\/span>\u00a0<\/span>The article brings together not only procedures that worked, but also the problems\u00a0encountered\u00a0during experimental development and extensive discussions of the solutions to these problems.<\/span>\u00a0<\/span>\u00a0<\/span><\/p>\n

This turns the protocol into a valuable tool for the scientific community.<\/span>\u00a0<\/span>Researchers\u00a0<\/span>who\u00a0wish<\/span>\u00a0to work with biocatalysis under similar conditions will now have a guide that\u00a0anticipates\u00a0difficulties, suggests solutions, and reduces the time needed to obtain reliable results.<\/span>\u00a0<\/span>As Itamar Neckel highlights,\u00a0<\/span>\u201c<\/span>it will be a very rich reference, because it contains both the measures that were carried out successfully,\u00a0<\/span>and<\/span>\u00a0also the problems that we faced<\/span>\u201d.\u00a0<\/span>\u00a0<\/span><\/p>\n

The protocol also provides guidance on critical steps, such as preliminary characterizations, radiation damage testing, sample preparation, and data acquisition<\/span>\u00a0\u2014\u00a0a<\/span>spects\u00a0that, if neglected, could compromise the entire experiment.<\/span>\u00a0<\/span>By\u00a0consolidating\u00a0this knowledge, the work\u00a0facilitates\u00a0new studies\u00a0and contributes to the advancement of a strategic area: the development of technologies based on natural processes to face global energy challenges.<\/span><\/p>\n

About LNLS<\/h2>

The Brazilian Synchrotron Light National Laboratory (LNLS) works with scientific research and technological development that involves synchrotron light, focusing on the operation and utilization of the multidisciplinary potential of Sirius, the country's most advanced scientific infrastructure. With ten research stations already online and open to the scientific and industrial communities, Sirius allows thousands of researchers from various areas to test their hypotheses about the microscopic mechanisms that produce the properties of both natural and synthetic materials which are used in a variety of fields such as health, the environment, energy, and agriculture. LNLS is part of the Brazilian Center for Research in Energy and Materials (CNPEM) in Campinas, S\u00e3o Paulo, a private, non-profit organization overseen by the Ministry of Science, Technology and Innovation (MCTI).<\/p>

https:\/\/lnls.cnpem.br\/<\/a><\/p><\/div>\n

About CNPEM<\/h2>

The Brazilian Center for Research in Energy and Materials (CNPEM) is home to a state-of-the-art, multi-user and multidisciplinary scientific environment and works on different fronts within the Brazilian National System for Science, Technology and Innovation. A social organization overseen by the Ministry of Science, Technology and Innovation (MCTI), with the involvement of the Ministry of Education and the Ministry of Health, CNPEM is driven by research that impacts the areas of health, energy, renewable materials, and sustainability. It is responsible for Sirius, the largest assembly of scientific equipment constructed in the country, and is currently constructing Project Orion, a laboratory complex for advanced pathogen research. Highly specialized science and engineering teams, sophisticated infrastructure open to the scientific community, strategic lines of investigation, innovative projects involving the productive sector, and training for researchers and students are the pillars of this institution that is unique in Brazil and able to serve as a bridge between knowledge and innovation. CNPEM's research and development activities are carried out through its four National Laboratories: Synchrotron Light (LNLS), Biosciences (LNBio), Nanotechnology (LNNano), Biorenewables (LNBR), as well as its Technology Unit (DAT) and the Ilum School of Science \u2014 an undergraduate program in Science and Technology.<\/p>

https:\/\/cnpem.br\/en\/<\/a><\/p><\/div>\n

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[vc_row][vc_column][vc_column_text css=””] Experimental protocol details challenges and solutions for the study of biocatalysts in beamlines with nanometric beams, such as Carna\u00faba, at Sirius A\u00a0work published in the\u00a0scientific\u00a0journal Nature Protocols\u00a0by researchers…<\/p>\n","protected":false},"author":28,"featured_media":65131,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":"","_links_to":"","_links_to_target":""},"categories":[1458,1481,1461],"tags":[],"class_list":["post-65133","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-releases-cnpem-en-2","category-releases-lnls-en","category-noticia-home-en-en","category-1458","category-1481","category-1461","description-off"],"acf":[],"yoast_head":"\nProtocol combines X-rays and electrochemistry to study hydrogen production - 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