This study evaluated the biogas production potential of water lettuce (Pistia stratiotes L.) by batch anaerobic digestion under in-vitro conditions. Twenty-one litre-plastic jars were used to conduct 4 replications over 75 days. The results showed that solution temperature, pH and Eh were suitable for biogas production. More than 50% of the obtained CH4 was formed within 17-42 days after incubation. The maximum daily CH4 production was 0.052 L/gVS, whilst the daily H2S concentration was low, with a maximum value of 28 ppm within 14 – 21 d after incubation. Moreover, the peak of daily biogas production was seen at day 16 with production of 0.12 L/gVS. The results highlight that water lettuce biomass can be potentially used to produce high quality biogas in anaerobic incubation.