Abstract
Biohydrogen production potential (BHP) depends on several factors like inoculum source, substrate, pH, among many others. Batch assays are the most common strategy to evaluate such parameters, where the comparison is a challenging task due to the different procedures used. The present method introduces the first internationally validated protocol, evaluated by 8 independent laboratories from 5 different countries, to assess the biohydrogen potential. As quality criteria, a coefficient of variation of the cumulative hydrogen production (Hmax) was defined to be <15 %. Two options to run BHP batch tests were proposed; a manual protocol with periodic measurements of biogas production, needing conventional laboratory materials and analytical equipment for biogas characterization; and an automatic protocol, which is run in a device developed for online measurements of low biogas production. The detailed procedures for both protocol options are presented, as well as data validating them. The validation showed acceptable repeatability and reproducibility, measured as intra- and inter-laboratory coefficient of variation, which can be reduced up to 9 %.
Highlights
Hydrogen production by dark fermentation is an emerging technology of increasing interest due to its renewable feature
Experimental design: Trial registration: Ethics: The proposed method presents two options to run Biohydrogen production potential (BHP) batch tests; a manual protocol with periodic measurements of biogas production; and an automatic protocol, which is run in a device developed for online measurements of low biogas production. n/a n/a
The assessment of the BHP is a common procedure to evaluate new potential substrates, and other parameters [1,2,3]. This method stands for the first standardized protocol validated internationally to estimate the BHP using batch tests
Summary
Hydrogen production by dark fermentation is an emerging technology of increasing interest due to its renewable feature. The thymolphthalein pH indicator in solution F will change from blue to colorless when the CO2 absorption capacity of the NaOH solution drops below the optimum At this point, it is recommended to replace the bottle with new solution F to prevent the CO2 gas from passing to the gas volume measuring device. 13 To change the solution F, place pipe plugs on the hoses of the incubation unit and gas measuring device. 7 Unplug the power adapters (for the motor controller and the gas volume measuring device) from the power supply. 9 Disconnect the hoses between the NaOH bottles and the gas volume measuring device. For cumulative hydrogen production results using the manual version, adjust the recorded volume to standard conditions (0 C and 1 atm).
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