Analysis of the diffusion of bile salt/phospholipid micelles in rat intestinal mucin.

The interaction of sodium taurocholate/egg phosphatidylcholine (TC/PC) micelles with mucin was determined to investigate the exclusion of lipids by mucus in the absorption process. The distribution of TC/PC was assessed at two intermicellar and three phospholipid concentrations with isolated, rat intestinal mucin (RIM) by dialysis. The diffusion coefficients were measured by NMR spectroscopy. At high [PC], RIM had lower [PC] relative to the control, while the concentration of TC was largely independent of mucin concentration. The PC diffusion coefficients were reduced in the presence of RIM. The magnetization decay of TC was compared with simulations to provide estimates of the monomeric diffusivity and exchange rate constant. The rate constants increased with increasing micelle concentration, and the free TC diffusion coefficient was reduced in the presence of mucin. Mucin has an exclusive effect on TC/PC mixed micelles that has been quantitatively determined through the use of diffusion measurements of dialyzed samples.

Interaction of bile salt and phospholipids with bovine submaxillary mucin.

PURPOSE: The purpose of this study was to determine the distribution and diffusion of sodium taurocholate-phospholipid micelles with mucin in order to provide the foundation for understanding the transport of ingested fat and poorly water-soluble drugs through the intestinal mucous layer. METHODS: Sodium taurocholate (NaTC) was dispersed with egg phosphatidylcholines (PC) to yield mixed micelles of a specific size and concentration. A preliminary study was conducted to determine the time required for equilibration of PC/TC micellar solutions with mucin. PC/TC micellar solutions were dialyzed against fixed and variable concentrations of bovine submaxillary mucin after which the concentration of PC and NaTC was measured by an assay for total phosphorus and by HPLC, respectively. In addition, a quantitative assay of TC and PC by NMR was developed and used to estimate the mobile fraction of lipids in the samples. Finally, pulsed-field gradient spin echo NMR self-diffusion measurements were made of the water, TC, and PC in the samples obtained from dialysis. RESULTS: TC/PC micellar solutions achieved equilibrium with mucin in 7 days. Mucin did not affect the equilibrium concentration of PC or TC, except at high concentrations of mucin (5%), and then the effect was small. NMR quantitation was valid for PC and TC systems containing small micelles, but deviated significantly with systems containing large micelles. Mucin decreased the diffusivity of water and the phospholipids, but the effect was relatively small. Mucin dramatically affected the mobility of TC, which prevented a straightforward interpretation of the calculated diffusion coefficients. CONCLUSIONS: Mucin has a minor effect on the equilibrium distribution of phospholipids and bile salts. However, lipids are readily accommodated by mucus, which can significantly increase the permeability of the mucous layer, particularly for poorly water-soluble drugs.

Distribution and diffusion of sodium taurocholate and egg phosphatidylcholine aggregates in rat intestinal mucin.

PURPOSE: The permeability of rat intestinal mucin (RIM) to sodium taurocholate/egg phosphatidylcholine (TC/PC)-mixed micelles has been investigated. METHODS: The time dependence for the equilibration of TC/PC-mixed lipid micelles with isolated RIM was determined. Thereafter the distribution of TC/PC-mixed lipid micelles was assessed at low and high PC and intermicellar concentrations (IMC) and with different RIM concentrations. The equilibrium distribution of PC and TC was determined by analysis for phosphorus and by high-performance liquid chromatography, respectively, as well as by nuclear magnetic resonance spectroscopy. In addition, the diffusion coefficients of water, PC, and TC were measured by pulsed field gradient nuclear magnetic resonance spectroscopy. Two model solutes, phenylmethyltrimethylsilane (PTMS) and tetramethylsilyl-tetradeutero-proprionic acid (TSP), were added to the high PC, low IMC samples, and the diffusion coefficients were determined. RESULTS: The time to reach equilibrium was 2 days for a system with a high intermicellar concentration of sodium taurocholate. At low PC concentrations, RIM had slightly higher PC concentrations relative to the control. In contrast, at high PC concentrations, RIM samples had lower PC concentrations. The concentration of TC was largely independent of mucin concentration. The water diffusivity was reduced proportionately to the concentration of RIM, and analysis indicated that about 150 g of water moved as a kinetic unit with each gram of mucin. The diffusion coefficients of PC were also reduced with increasing RIM concentration. The magnetization decay of TC did not always follow a monoexponential decay, reflecting the simultaneous diffusion and exchange among sites imparting different relaxation behavior on the TC. Magnetization decay curves were simulated and the diffusivity of TC in mucin was estimated. The diffusion coefficient of TSP was 10 times larger than that of PTMS in the presence of micelles and mucin. CONCLUSIONS: RIM is highly hydrated, and dilute solutions have a minor exclusive effect on the high concentration of PC/TC micelles. At low concentrations of PC, there appears to be preferential association of the PC with the RIM. The permeability of mucin to solutes in the presence of bile salt mixed micelles critically depends on the degree of association of the solute with the micelle.