Poly(ethylene glycol)(PEG)-lipid anchor conjugates can prolong the circulation lifetimes of liposomes following intravenous injection. In this work we investigate the influence of the lipid anchor and the nature of the chemical link between the PEG and lipid moieties on circulation lifetime. It is shown that incorporation of N-(monomethoxypoly(ethylene glycol)2000-succinyl)-1-palmitoyl-2-oleoylphosphatidylethanolamide (MePEG2000-S-POPE) into large unilamellar vesicles (LUVs) composed of distearoylphosphatidylcholine (DSPC) and cholesterol (DSPC/cholesterol/MePEG2000-S-POPE, 50:45:5, mol/mol) results in only small increases in the circulation lifetimes as observed in mice. This is shown to be due to rapid removal of the hydrophilic coating in vivo, which likely arises from exchange of the entire PEG-lipid conjugate from the liposomal membrane, although chemical breakdown of the PEG-lipid conjugate is also possible. The chemical stability of four different linkages was tested, including succinate, carbamate and amide linkages between MePEG derivatives and the amino head group of PE, as well as a direct link to the phosphate head group of phosphatidic acid (PA). The succinate linkage was found to be the most labile. The anchoring capability of DSPE as compared to POPE in PEG-PE conjugates was also examined. It is shown that incorporation of MePEG2000-S-DSPE conjugates into DSPC/cholesterol LUVs results in little loss of the PEG coating in vivo, long circulation lifetimes and reduced chemical breakdown of the PEG-lipid conjugate. This work establishes that DSPE is a considerably more effective anchor for PEG2000 than POPE and that the chemical stability of PEG-PE conjugates is sensitive to the nature of the linkage and exchangeability of the PEG-PE complex. We suggest that retention of the PEG coating is of paramount importance for prolonged circulation lifetimes.