The pending global energy crisis is stimulating the quick growth of biodiesel manufacturing. However, biodiesel production cost is increasing due to the accumulation of crude glycerol as a by-product. With a ton of biodiesel produced, the process theoretically yields 100 kg of crude glycerol (contaminated glycerol) as a by-product. The objective of this project is to convert crude glycerol from biodiesel production to bio-hydrogen by a biological method suitable for industries in Thailand. A mix culture of microorganisms from wastewater treatment process is used to anaerobically digest crude glycerol in an eight liter-packed-bed reactor. The operation is semi-batch under atmospheric temperature and pH is controlled. Crude glycerol is fed every other day when gas and liquid products are sampled and analyzed by GC-TCD and GC-FID, respectively. The results conclusively show that glycerol content is reduced and hydrogen gas is generated with alcohol products. Currently, the operating conditions are being varied to study their effects on the hydrogen production.