In the near future, crude oil based fuels must little by little be replaced by biofuels both in the region of the European Union (EU) and in the United States. Bearing this in mind, a Finnish-made off-road diesel engine was tested with a biofuel-diesel fuel blend in the Internal Combustion Engine (ICE) Laboratory of Turku Polytechnic, Finland. The biofuel was cold-pressed mustard seed oil (MSO). The engine operation, performance and exhaust emissions were investigated using a blend of 30 mass-% MSO and 70 mass-% diesel fuel oil (DFO). The injection timing of the engine was retarded considerably in order to reduce NOx emissions drastically. The main target was then to find out, whether the blended oxygen containing MSO would speed up the combustion so that the particulate matter (PM) emissions would remain unchanged or even decrease despite the injection retardation. As secondary tasks of the study, the NOx readings of the CLD and FTIR analyzers were compared, and exhaust contents of unregulated compounds were determined. Retarding the injection timing resulted in a significant decrease of NOx emissions, but in an increase in smoke, as expected. At retarded timing, the NOx emissions remained almost unchanged, but the amount of smoke decreased when the engine was run with the fuel blend instead of DFO. At retarded timing at rated speed, the number of ultra-fine particles decreased, but the amount of large particles increased with DFO at full load. At 10% load, however, the particle number increased in the entire particle size range due to retardation. At both loads, the use of the fuel blend slightly reduced larger particles, whereas the number of small particles somewhat increased. At full load at an intermediate speed of 1500 rpm, the PM results were very similar to those obtained at rated speed. At 10% load with DFO, however, the injection retardation led to a higher number of larger particles, the smaller particles being at almost an unchanged level. With the fuel blend, the particle number was now higher within almost the whole particle diameter range than with DFO. Considerably higher NO2 contents were usually detected with FTIR than with CLD. The shape of the NOx result curves were rather similar independent of which one of the analyzers was used for measurements. The NOx contents were, however, generally some ten ppms higher with FTIR. The exhaust contents of unregulated compounds were usually low.