The purpose of this study was to develop oil-in-water emulsions (100-120 nm in diameter) and to correlate the surface properties of the emulsions with blood residence time and accumulation into neoplastic tissues by passive targeting. We investigated the effect of phospholipid and sphingolipid emulsifiers, hydrogenated soybean phosphatidylcholine (HSPC) and egg sphingomyelin (ESM), in combination with polysorbate 80 (PS-80) and 1,2-distearoyl-sn-glycero-3-phosphatidylethanolamine (DSPE)-PEG lipids of various PEG chain lengths and structures in prolonging circulation time and enhancing accumulation into B16 melanoma or C26 colon adenocarcinoma. The relationship between amphiphile molecular packing at the air/water interface on emulsion stability upon dilution in albumin and circulation longevity in vivo was also explored for non-PEGylated emulsions. PEGylation of the droplet surface with 10-15 mol% of DSPE-PEG 2000 or 5000 enhanced the circulation time of the emulsions, however, accumulation was only observed in the C26 tumor model. The tighter molecular packing observed with ESM/PS-80 monolayers at the air/water interface compared to HSPC/PS-80 correlated with improved emulsion stability in vitro, however, enhanced circulation time in vivo was not observed. A better understanding of the relationships between composition and performance will result in improved emulsion-based drug delivery vehicles for cancer therapy.