Beet molasses and black strap are two major waste streams of the sugar production industry. They both contain high amounts of sucrose which makes them suitable substrates for biological hydrogen production. Photofermentation has the potential capacity to effectively use a variety of feed stocks. A comparative study on photofermentative biohydrogen production from beet molasses, black strap and sucrose was conducted. With yields of 10.5 mol H2/mol sucrose for beet molasses (1 g/L sucrose); 8 mol H2/mol sucrose for black strap (1 g/L sucrose) and 14 mol H2/mol sucrose for pure sucrose, a one stage photofermentation system seems to be a good alternative to two-stage systems given the potential savings in energy input and operational costs.