Biohydrogen production from tequila vinasses using a xed bed reactor Germán Buitrón, Dorian Prato-Garcia and Axue Zhang ABSTRACT In Mexico, the industrial production of tequila leads to the discharge of more than 31.2 million of m 3 of vinasse, which causes serious environmental issues because of its acidity, high organic load and the presence of recalcitrant compounds. The aim of this research was to study the feasibility of a xed bed reactor for the production of biohydrogen by using tequila vinasse as substrate. The experiments were carried out in a continuous mode under mesophilic and acidic conditions. The maximum hydrogen yield and hydrogen production rate were 1.3 mol H 2 mol/mol glucose and 72 ± 9 mL H 2 /(L reactor h), respectively. Biogas consisted of carbon dioxide (36%) and hydrogen (64%); moreover methane was not observed. The electron-equivalent mass balance tted satisfactorily (sink of electrons from 0.8 to 7.6%). For vinasses, hydrogen production accounted for 10.9% of the total available electron-equivalents. In the liquid phase, the principal metabolites identied were acetic, butyric and iso-butyric acids, which indicated a butyrateacetate type fermentation. Tequila vinasses did not result in potential inhibition of the fermentative process. Considering the process as a water treatment system, only 20% of the original carbon was removed (as carbon dioxide and biomass) when the tequila vinasses are used. Germán Buitrón (corresponding author) Dorian Prato-Garcia Axue Zhang Laboratory for Research on Advanced Processes for Water Treatment, Unidad Académica Juriquilla, Instituto de Ingeniería, Universidad Nacional Autónoma de México, Blvd. Juriquilla 3001, Querétaro 76230, México E-mail: GbuitronM@ii.unam.mx Key words | biohydrogen, continuous reactor, dark fermentation, xed bed reactor, vinasses INTRODUCTION Vinasse is the dark-colored wastewater generated after the distillation of ethanol that is produced during the fermenta- tion of sugars. From 10 to 12 L of vinasses are generated as residue per litre of tequila produced (Espinoza-Escalante et al. ; Carvajal-Zarrabal et al. ). In 2013, there were produced in Mexico about 226 × 10 6 L of tequila (CRT ). This residue poses a signicant disposal and treatment problem because of low pH (34), high temperature (70 90 W C), high biological oxygen demand (3060 g/L), and the presence of dissolved salts, melanoidins and polyphenolic substances (Buitrón & Carvajal ; Carvajal-Zarrabal et al. ). Therefore, researchers have been focusing on improving the potential of biological treatment of this waste. Sugarcane and tequila vinasse have been satisfactorily treated by coagulationocculation, electrocoagulation, anaerobic digestion and advanced oxidation processes (Carvajal-Zarrabal et al. ; Zampol-Lazaro et al. ). Anaerobic digestion is the most common method currently applied in industrial scale. However, the high chemical oxygen demand (COD) content in vinasses makes the process inefcient and leads to a large amount of sludge. In another common way, raw vinasse is applied as a fertili- zer (fertirrigation) in sugarcane crops, which represents a low-cost alternative to prevent the pollution of water bodies. Fertirrigation has led to an increase of crop yield and to reduction of the usage of chemical fertilizers. How- ever, fertirrigation can cause salinization and soil erosion, lixiviation of metals, alkalinity reduction, phyto toxicity and unpleasant odor (Pant & Adholeya ; Christofoletti et al. ). Moraes et al.() suggest that sugarcane vinasse has great potential for energy production, but this potential is wasted because of its application as a fertilizer. The authors pointed out that the energy wasted by a sugar- cane mill that uses fertirrigation as an alternative to treat vinasse may be equivalent to the electricity demand of a city of 130,000 inhabitants. Biohydrogen is a promising fuel since it is considered as clean, renewable, and high energy density (122 kJ/g) energy. It has been illustrated that biohydrogen can be produced efciently from sugarcane vinasse, tequila vinasse, 1919 © IWA Publishing 2014 Water Science & Technology | 70.12 | 2014 doi: 10.2166/wst.2014.433