The relationship between ethanol and iso-butanol fuel concentrations and vehicle particulate matter emissions was investigated. This study utilized a gasoline direct injection (GDI) flexible fuel vehicle (FFV) with wall-guided fueling system tested with four fuels, including E10, E51, E83, and an iso-butanol blend at a proportion of 55% by volume. Emission measurements were conducted over the Federal Test Procedure (FTP) driving cycle on a chassis dynamometer with an emphasis on the physical and chemical characterization of particulate matter (PM) emissions. The results indicated that the addition of higher ethanol blends and the iso-butanol blend resulted in large reductions in PM mass, soot, and total and solid particle number emissions. PM emissions for the baseline E10 fuel were characterized by a higher fraction of elemental carbon (EC), whereas the PM emissions for the higher ethanol blends were more organic carbon (OC) in nature. The higher ethanol blends and the iso-butanol blend showed lower concentrations of accumulation mode particles and size distributions shifted to smaller particle sizes compared to E10. In addition, the majority of trace elements and metals showed clear reductions with increasing alcohol content into gasoline.