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It has been possible to prepare electrostatically stabilized aqueous dispersions of lipid A (diphosphate) particles of low polydispersity at low ionic strength (1-10 mM NaCl) over a range of volume fractions of 1.5 × 10-4 < < 5.75 × 10-4 (25 C). These suspensions have been characterized by transmission electron microscopy, light scattering, osmotic pressure measurements, and small-angle X-ray scattering experiments at 25 C. All four measurements yielded independent values for particle sizes, weighted-average molecular weights, number-average molecular weights, and particle surface charge. The mean values obtained are = 37.59 ± 0.75 nm, = 24.89 ± 0.88 nm, = (10.55 ± 0.78) × 106 g/mol, = (9.81 ± 0.90) × 106 g/mol, and the effective surface charge Z* = (756 ± 85). Very good experimental agreement is found for the directly measured osmotic pressure values and those determined from light scattering and small-angle X-ray scattering measurements as a function of volume fraction, , by applying liquid-state theory models. Using the particle parameters for the lipid A (diphosphate) system as determined, the scattering functions and the osmotic pressures can be compared as a function of volume fraction with no adjustable parameters. The ordering of lipid A in solution revealed a body-centered cubic (bcc) type lattice (a = 36.14 nm) at volume fractions of 3.75 × 10-4 < < 4.15 × 10-4, whereas at volume fractions of 4.15 × 10-4 < < 5.75 × 10-4 in the presence of 1.0 mM NaCl a face-centered cubic (fcc) lattice type (a = 57.25 nm) was observed. Small-angle X-ray scattering experiments also indicate the presence of long-ranged order at 1.0 mM or at 10.0 mM NaCl for lipid A dispersions of 3.75 × 10-4 < < 5.75 × 10-4.

Electrostatically stabilized aqueous dispersions of nm-sized free lipid A particles at low volume fractions (1.0×10-4 ρTEM=8.0±0.6%) yield similar values for the particle size and particle size distribution ( ρQELS=10.9±0.75 %), whereas conductivity and acid-base titrations estimate surface chemical parameters (dissociation constant, ionizable sites, and Stern capacitance). Effective particle charges were determined by fits of the integral equation calculations of the polydisperse static structure factor, S(Q), to the light scattering data. Using the particle properties as determined from these experiments, the polydisperse structure factor, S(Q), was calculated as a function of volume fraction, θ, which was found to be consistent with a S(Q) dependence on the number particle density. It can be concluded that, at low volume fractions and low ionic strength, the light scattering data are well represented by a Poisson-Boltzmann model (PBC) of fluid-like ordering of free lipid A in aqueous solution. We find that the light scattering data of this dispersion are best described by a model where only a small fraction of the ionizable phosphate groups is dissociated at neutral pH. Finally, light scattering studies of lipid A dispersions of volume fractions of 3.9×10-4