Diminishing reserves of crude oil across the globe has led to a shift towards the alternate methods of energy production in order to meet the energy demands for the growing population of the world. Amongst the many promising areas, bio-crude production from 1st, 2nd, 3rd, and 4th generation biomasses is a promising one because of its versatility, mass availability of potential feedstocks, and sustainability. Biomasses are converted through different chemical, bio-chemical and thermo-chemical processes into biocrude and other value-based necessary products. However, this biocrude oil needs to be further upgraded in order to be used as a drop in fuel. Not all biocrude produced from these methods and resources has the same standard properties that are required for it to be used as a drop in fuel. Standard requirements like viscosity, density, stability, energy density, combustion, thermal and elemental properties etc. depend on the type of feedstock used and thus need to be checked and evaluated to produce finished fuels that comply with regulated specifications. This chapter deals with these analytical techniques employed for the physical, chemical and elemental analysis of bio fuels. Moreover, fuel property evaluation and international standards & specifications are also discussed to compare the quality of the bio fuels with the regulatory standards.