Background: The ability for hydrogen production of 13 native strains of Clostridium spp. strains isolated from Colombian soil was evaluated using glycerol substrate. Glycerol to hydrogen conversion was investigated using a batch fermentation reactor and industrial glycerol source (50 g.l-1, pH 7.00). Results: The results were quantified regarding acids, hydrogen, biomass and solvent production. The selected strain gave good hydrogen over production output at 14.4 mmol H2.l-1, productivity 0.3167 mg H2.h-1 l-1 culture mediumand yield 0.1962 mol H2.mol-1 glycerol. A further fermentation assay a 4.0 liter batch reactor let to being 0.26 mg.l-1.h-1 after 18 hrs of fermentation. Logistic model, Luedeking-Piret model and Luedeking-Piret modified models were used for modeling changes in cell growth, hydrogen production and substrate consumption (Correlation coefficients R² = 0.95 for biomass substrate, R² = 0.77 hydrogen production). Conclusions: Our results indicate that hydrogen production through glycerol bioconversion by native strains is possible.