Mitigating Microbial Artifacts in Laboratory Research on Underground Hydrogen Storage

The global energy sector is aiming to substantially reduce CO2 emissions to meet the UN climate goals. Among the proposed strategies, underground storage solutions such as radioactive disposal, CO2, NH3, and underground H2 storage (UHS) have emerged as promising options for mitigating anthropogenic emissions. These approaches require rigorous research and development (R&D), often involving laboratory-scale experiments to establish their feasibility before being scaled up to pilot plant operations. Microorganisms, which are ubiquitous in laboratory environments, can significantly influence geochemical reactions under variable experimental conditions of porous media and a salt cavern. We have selected a consortium composed of Bacillus sp., Enterobacter sp., and Cronobacter sp. bacteria, which are typically present in the laboratory environment. These microorganisms can contaminate the rock sample and develop experimental artifacts in UHS experiments. Hence, it is pivotal to sterilize the rock prior to conduct experimental research related to effects of microorganisms in the porous media and the salt cavern for the investigation of UHS. This study investigated the efficacy of various disinfection and sterilization methods, including ultraviolet irradiation, autoclaving, oven heating, ethanol treatments, and gamma irradiation, in removing the microorganisms from silica sand. Additionally, the consideration of their effects on mineral properties are reviewed. A total of 567 vials, each filled with 9 mL of acid-producing bacteria (APB) media were used to test killing efficacy of the cleaning methods. We conducted serial dilutions up to 10−8 and repeated them three times to determine whether any deviation occurred. Our findings revealed that gamma irradiation and autoclaving were the most effective techniques for eradicating microbial contaminants, achieving sterilization without significantly altering the mineral characteristics. These findings underscore the necessity of robust cleaning protocols in hydrogeochemical research to ensure reliable, reproducible data, particularly in future studies where microbial contamination could induce artifacts in laboratory research.