The helicopter from the Air Test and Evaluation Squadron Two One used a fuel mixture made from camelina seed, which is in the same family of plants as the mustard seed and rapeseed.
Camelina needs little water or nitrogen to flourish and can be grown on marginal agricultural soil.
"These biofuels provide the Navy with an 'off-ramp' from petroleum to increased energy security," said Rear Adm. Philip Cullom, director, Navy Task Force Energy.
Rick Kamin, the Navy Fuels lead, said the helicopter tests will further demonstrate this fuel made from an alternative, non-petroleum feed stock is a viable option for use in Navy aircraft.
The Navy earlier this year tested the biofuel blend on the F/A-18 Super Hornet. Results from those tests indicated the aircraft performed as expected through its full flight envelope with no degradation of capability.
Testing will continue across additional aircraft models in 2011 with a target of approving the 50/50 biofuel blend for use in the Navy ships and aircraft by early 2012.