In this work, we developed a passive millimeter-micrometer fluidic device for biodiesel production which is composed of the esterification, purification and separation sub-systems. The biodiesel produced by this device was confirmed the FAME standard ASTM D6751 in U.S.A.. While the raw material (soybean oil and methanol) go through the esterification system, the conversion rate is more than 90% at room temperature (23 oC). Then, the methanol content of the biodiesel was reduced from 6% before washing to less than 0.2% after washing. Eventually, the pure biodiesel with no glycerol peaks in the NMR chart was collected from the upper outlet of this chip. This millimeter-micrometer fluidic device integrated the continuous biodiesel production processes with no stirring, heating and any additional instruments by passive structure of flow channels which is attainable, conforming to the requirements of the global trend for green energy development.