The real-time analysis of volatiles (primary tar) produced during the fast pyrolysis of biomass in a microfluidized bed reactor (MFBR) is achieved by online single photoionization mass spectrometry (SPI-MS). The effect of biomass composition (Douglas fir, oak, and miscanthus), particle shape and size (cylinder, lamella, or powder), bed temperature, and fluidizing gas flow-rate on primary tar composition is studied. Principle component analysis is conducted on the major ions analyzed by SPI-MS to evidence the significant differences between conditions. The variance in obtained SPI-MS spectra reveals the important effect of biomass composition and temperature on volatiles composition. The effect of particle size on volatiles composition is clearly evidenced. Typical pyrolysis regimes are defined according to specific markers which are key chemical compounds to characterize biomass fast pyrolysis. SPI-MS combined with a MFBR is an interesting tool to unravel the effects of biomass composition and of heat and mass transfers on biomass fast pyrolysis processes.