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Physical Chemistry of Bile

Donna J. Cabral, Donald M. Small

10.1002/cphy.cp060331

Source: Supplement 18: Handbook of Physiology, The Gastrointestinal System, Salivary, Gastric, Pancreatic, and Hepatobiliary Secretion

Originally published: 1989

Published online: January 2011

Full Article on Wiley Online Library



Abstract

The sections in this article are:

1 Structural and Surface Properties of Bile Salts
1.1 Molecular Structure
1.2 Crystal Structure
1.3 Surface Chemistry
1.4 Hydrophobic‐Hydrophilic Balance
2 Aqueous Solution Properties of Bile Salts
2.1 Solubility in Water
2.2 Solubility in Organic Solvents
2.3 Ionization Behavior
2.4 Aggregation Behavior
3 Bile Salt–Lecithin Interactions
3.1 Composition of Lecithins in Bile
3.2 Bile Salt–Phospholipid–Water Systems
4 Cholesterol
4.1 Solubility in Aqueous Media
4.2 Cholesterol‐Bile Salt Interactions
4.3 Cholesterol‐Phospholipid Interactions
4.4 Quaternary System: Bile Salt‐Phospholipid‐Cholesterol‐Water
5 Bile
5.1 Normal Bile
5.2 Metastable Bile
5.3 Cholesterol Crystal Formation in Bile
5.4 Secretion of Bile from the Hepatocyte
Figure 1. Figure 1.

Molecular structure of common bile acids showing common steroid ring and side‐chain structure. Hydroxyl group(s) location and orientation are given for each bile acid.
Figure 2. Figure 2.

Crystal structure of bile acids and salts. A: stereoview of the cell of lithocholic acid: α‐axis projection with c‐axis vertical. Oxygen molecules are blackened. B: stereoview of chenodeoxycholic acid; b‐axis projection. C: stereoview of unit cell of deoxycholic acid: H2O, showing channel filled with H2O molecules. Oxygens of H2O molecules are blackened. D: crystal packing of DCA: Rb viewed along b‐axis, showing planar wavy bilayer patterns. Broken lines, hydrogen bonds; large closed circles, Rb+, smaller closed circles, oxygen from H2O; small open circles, oxygen from the bile acid. E: crystal packing of unit cell of Na‐cholate‐monohydrate b‐axis projection. Wavy bilayer pattern is illustrated. Broken lines, hydrogen bonds; larger closed circles, Na+; smaller closed circles, carbon atoms; open circles, oxygen atoms. F: stereoview of bilayer packing of calcium cholate chloride heptahydrate. Closed circles of cholate molecules represent oxygen atoms of hydroxyl and carboxyl groups. Closed circles between cholate molecules represent Ca2+. G: stereoview of bilayer packing of sodium cholate monohydrate. Closed circles of cholate molecules represent oxygen atoms of hydroxyl and carboxyl groups. Closed circles between molecules represent H2O. H: stereoview of bilayer packing of rubidium deoxycholate. Closed circles of deoxycholate molecules represent oxygen atoms of hydroxyl and carboxyl groups. Closed circles between molecules represent H2O.

A from Arora et al. 4; B from Lindley et al. 68; C from Tang et al. 128; D from Coiro et al. 30; E from Cobbledick and Einstein 29; F–H from Hogan et al. 48a)
Figure 3. Figure 3.

Schematic of surface configuration of cholanic acid and hydroxylate derivatives. Maximum area per molecule was taken from first inflection point of compression isotherms; minimum area was taken from collapse point. Pressure at film collapse and state of film are given. Collapse pressure for chenodeoxycholic acid in 3 M NaCl low pH was determined to be ∼27 dyn/cm, similar to that of deoxycholic acid.

Data from Carey 20 and Small 113 and references therein
Figure 4. Figure 4.

Reverse‐phase high‐performance liquid chromatography (HPLC) chromatographs of mixture of bile salts. For a given conjugation state, mobility decreases in the order UDCA > CA > CDCA > DCA. Column used was an Ultrasphere ODS 250 X 4.6 mm, with a mobile phase of 75% MeOH‐25% 0.005 M KH2PO4/H3PO3, pH 5.0. TUDC, tauroursodexoycholate; TC, taurocholate; TCDC, taurochenodeoxycholate; TDC, taurodeoxycholate; GUDC, glycoursodeoxycholate; GC, glycocholate; GCDC, glycochenodeoxycholate; GDC, glycodeoxycholate; UDC, ursodeoxycholate; C, cholate; CDC, chenodeoxycholate; DC, deoxycholate. OD, optical density.

From Armstrong and Carey 3
Figure 5. Figure 5.

Hypothetical titration curve for solutions of free bile salts or for glycine conjugates. W, first equivalence point where titration of bile salt with hydrochloric acid commences. Y, last point where bile salt solution is in thermodynamic equilibrium as a single aqueous phase. T, Tyndall effect noted in this region of titration curve. X, point where precipitation of bile acid crystals commences. X', equilibrium pH at point of bile acid precipitation. Z, second equivalence point where titration of bile salt with hydrochloric acid is complete. TOT, total amount of acid required to complete the titration. HA, amount of acid added from first equivalence point W to point Y, which represents maximum solubility of bile acid (HA) in bile salt solution (A‐).

From Small 113
Figure 6. Figure 6.

Titration curves for cholic acid (CA) in several molecular environments. The pH is changed by adding HCl or NaOH, and chemical shift of carboxyl carbon (C‐24) is monitored by 13C NMR. Aqueous monomeric CA (.2 mM below its CMC and solubility limit). Aqueous micellar CA 116 mM). CA‐sodium taurocholate mixed micelles 1:7 mol/mol). CA‐BSA (bovine serum albumin) complexes 2:1 mol/mol). CA–egg yolk lecithin vesicles 1:20 mol/mol), major peak from CA in outer monolayer of vesicle, minor peak from CA in inner monolayer of vesicle.

Adapted from Cabral et al. 15
Figure 7. Figure 7.

Schematic of structure of bile salt micelles. Molecules of primary micelles associate via hydrophobic regions with aggregation numbers from 2 to 10. At higher concentrations some bile salts form larger aggregates, secondary micelles, stabilized by hydrophobic associations and hydrogen bonding.

From Small 112, printed with permission from Advances in Chemistry Series, copyright 1968, American Chemical Society
Figure 8. Figure 8.

Lecithin–sodium cholate–water diagram. W, H2O; L, lecithin; NaC, sodium cholate. Along side W‐L, numbers indicate H2O wt percentage; along side L‐NaC, numbers indicate lecithin wt percentage; along side NaC‐W, numbers indicate NaC wt percentage. Solid lines, well‐defined boundaries (+L2%); broken lines, less well‐defined frontiers. I, Zone of neat phase. II, Zone of cubic phase. III, Zone of middle phase. IV, Zone of isotropic phase. 1, Zone of separation of neat phase and NaC crystals. 2, Zone of separation of cubic phase and NaC crystals. 3, Zone of separation of middle phase and isotropic solution. 4, Zone of separation of neat phase and cubic phase. 5, Zone of separation of cubic phase and middle phase. 6, Zone of separation of neat phase and middle phase. 7, Zone of separation of neat phase and isotropic phase. 8, Zone of separation of isotropic solution and NaC crystals, a–e, Zones of separation of 3 phases whose compositions for each zone are indicated by apices of the triangles. Points A–H, J, K, M, O–Q are discussed in text. Right side of diagram, molar ratios of lecithin to NaC.
Figure 9. Figure 9.

Lecithin–sodium cholate water diagram at high‐H2O concentrations. NaC, sodium cholate; W, H2O. Left: larger numbers, wt%, smaller numbers, concentrations (mM). Parts of single‐phase zones I, III, and IV are shown, as well as 2‐phase regions 3, 6, and 7. Approximate tie lines in regions 6 and 7 and 3‐phase region are illustrated. Right, Bragg spacings (D) from lamellar and hexagonal lattices. These lattice parameters are given in angstroms along high‐H2O phase boundaries of lamellar phase and hexagonal phase. Dashed line from P (critical micellar concentration of NaC in H2O) to O, separates micelles formed by dilution of hexagonal phase (below) from micelles formed by dissolution of NaC crystals (above).
Figure 10. Figure 10.

Lecithin–sodium cholate system at >90% H2O, illustrating different techniques for preparing mixtures. NaC, sodium cholate; W, H2O. NaC‐W axis: larger numbers, wt% NaC from 0% to 10%; smaller numbers, concentration (mM). Point P, critical micelle concentration of NaC in H2O (.52% or ∼12 mM). Lecithin axis: large numbers, wt% lecithin; small numbers, concentrations (mM). Zone IV has been divided into 2 regions: IVa and IVb. Line separating 2 regions is a straight line running from point P to point O (see Fig. 9 and indicates a change in micellar structure. Narrow 2‐phase region 3 and larger 2‐phase region 7 are spearated by 3‐phase region a. Heavy lines, various methods of preparing mixtures. Any mixture may be made up by coprecipitation method (e.g., points 1 on 10% solid line and 5% solid line; TS, total solutes). A mixture made up by coprecipitation may then be diluted with H2O or buffer and will follow line 2. If a coprecipitated mixture is diluted with a specific concentration of bile salt, e.g., 2%, it will follow line 3. A coprecipitated mixture such as that plotted on 5% line at 40 mM lecithin and 45 mM NaC may be placed in a dialysis bag and dialyzed against H2O, buffer, or a bile salt solution. Because lecithin does not escape from the dialysis bag and its concentration remains unchanged, the composition will move toward W‐lecithin side along a constant 40‐mM line, line 4. Concentration of dialysate (not shown) will be found along W‐NaC axis. Finally, any concentration of lecithin, eg., a 6% suspension (either coarse liposomes or unilamellar vesicles), may be diluted with a specific concentration of bile salt, for instance at 10%, as shown on line 5. Compositions will fall on line 6, depending on amount of 10% bile salt solution added.
Figure 11. Figure 11.

A: mean hydrodynamic radii () of sodium taurocholate‐lecithin solutions (L/NaTC) at various total lipid concentrations (.625%–10% at 24°C in 0.15 M NaCl). diverges as lecithinbile salt ratio approaches micellar‐phase limits, and divergence limits are very concentration dependent. Dashed line, radius of simple discoid micelle. Phase limits shown at bottom of each panel represent appropriate lecithin‐bile salt ratio for total lipid concentrations of a, 0.0625%; b, 1.25%; c, 2.5%; d, 5.0%; and e, 10.0%. values indicated at tops of certain curves represent size of bilayer vesicles formed in supersaturated systems. B: at 3 L‐NaTC ratios as a function of the total wt% of lipids. As the total percentage of lipids increases, decreases markedly and approaches the dimensions of the simple discoidal micelle (arrows at right) at high lipid concentrations.

A reprinted with permission from Mazer et al. 70. Copyright 1980 American Chemical Society
Figure 12. Figure 12.

Lecithin‐sodium taurocholate (NaTC) system at high H2O (W) content. A: divergence limit (see Fig. 11A) and coexistence boundary plotted on rectangular coordinates. B‐C: system plotted on triangular coordinates (note difference in scale). Dashed line, phase boundary between micellar phase and zone 3. Phase boundary is virtually same as divergence limit line. Point P (critical micellar concentration of NaTC in H2O) has a value of 3.2 mM; 2‐phase regions 3 and 7 are shown. Tie lines in region 7 and the 3‐phase region a are also shown.

A from Mazer et al. 70; B‐C, coexistence and divergence boundaries from Mazer et al. 70
Figure 13. Figure 13.

A: mean hydrodynamic radius . B: polydispersity index V of a mixed micellar stock solution sodium glycocholatelecithin [lecithin/sodium glycocholate = 0.85; total concentration (Ctot) = 5 g/dl; temperature = 20°C] as a function of dilution. Dilution factor of 1 corresponds to stock solution Ctot, and dilution factor of n corresponds to Ctot divided by n.

From Schurtenberger et al. 108. Reprinted with permission from Journal of Physical Chemistry, copyright 1985, American Chemical Society
Figure 14. Figure 14.

Lecithin‐sodium glycocholate‐water system in dilute region of zone 7. L, lecithin; NaGC, sodium glycocholate; W, water. Closed circles, composition of fluid within dialysis bag; open circles, corresponding compositions (connected by a line) of dialysate fluid. Lines connect aqueous phase (intermicellar concentration or dialysate) with mixture composition (closed circles) for a series of vesicles of different sizes [ (Å)]. Extensions of these lines intersect L‐NaGC border (as shown by closed circles on right) and give ratio of NaGC bound to lecithin vesicle. Rh, mean hydrodynamic radii.
Figure 15. Figure 15.

Rates of dissolution of multilamellar liposomes by bile salts (BS) of different concentration. Lecithin concentrations of 1.5 mM were diluted with an equal volume of bile salts at concentrations on horizontal axis. A: time for half dissolution (T1/2 is plotted as a function of bile salt concentration; B: complex rate constant k1 91 for the process is plotted. Order of dissolution rates, chenodeoxycholate (CDC) > deoxycholate (DC) > cholate (C) > ursodeoxycholate (UDC).
Figure 16. Figure 16.

Sodium taurocholate‐lecithin‐cholesterol phase diagram. Chol, cholesterol; NaTC, sodium taurocholate; PL, phospholipid. At 10% total lipid, micellar phase is bounded by solid line. Dotted lines, micellar boundaries with decreasing concentrations of total lipids. Liquid crystal phase is a phospholipid phase with varying amounts of cholesterol and small amounts of bile salts. Crystalline phase is cholesterol monohydrate. We suggest that a 3‐phase region cannot directly abut on a 1‐phase region, except at a point where the 2‐phase region on the right extends to point B. The 3‐phase region consists of 1) cholesterol crystals, 2) liquid crystals of phospholipid, cholesterol, and bile salt, and 3) a micellar solution of composition B.

Adapted from Carey et al. 24
Figure 17. Figure 17.

Phase diagrams of aqueous taurine‐conjugated ursodeoxycholic acid (TUDC)‐, cholic acid (TC)‐, or chenodeoxycholic acid (TCDC)‐lecithin‐cholesterol monohydrate at a total lipid concentration of 10 g/100 ml (.20 M Na+, pH 7.4, 37°C). Axes are expressed in mol%. Although phase diagrams show 3‐phase region abutting 1‐phase region, there must be a 2‐phase region, however small, separating them.

From Salvioli et al. 101
Figure 18. Figure 18.

Partial phase diagram for aqueous system sodium taurocholate‐lecithin‐cholesterol 20 g/100 ml, 0.15 M NaCl, 24°C). Shaded area, metastable region; open circles, maximum equilibrium cholesterol solubility; closed circles, metastable‐labile limit.

From Carey et al. 24
Figure 19. Figure 19.

Behavior of hypothetical biles of compositions a‐f as they transform from dilute hepatic bile to concentrated gallbladder bile. Total lipids for hepatic bile are ∼1.5% and for gallbladder bile 10%. Phase boundaries are from Carey et al. 24. BS, bile salt. Point B is the same as that discussed in Fig. 16. For further explanation, see text.
Figure 20. Figure 20.

Electron micrograph of a bile canaliculus of a bile salt‐depleted rat liver that shows the presence of vesicular material within the lumen (left) (X 32,800). Higher magnification (inset) illustrates more clearly unilamellar structure of vesicle and can be compared with structure of canalicular membrane. Mean diameter of these vesicles is 56 nm 30–85 nm). Similar vesicles are also apparent in fresh bile specimen of same animal (right). Bars, 0.25 μm.

From Ulloa et al. 130, © by Am. Assoc. of the Study of Liver Diseases, 1987
Figure 21. Figure 21.

Distribution and composition of biliary lipids in bile fractionated on a continuous metrizamide density gradient. Left, T‐tube human hepatic bile; right, rat hepatic bile. Histograms are mean +L SD of percentage of each lipid in total sample of 7 human biles and 4 rat biles. Total lipid composition of rat biles was 2.4 g/dl; total composition of human T‐tube biles was 1.7 g/dl. BS, bile salt; C, cholesterol; PL, phospholipid. Compositions of each of the density cuts are plotted below on triangular coordinates with appropriate line as determined by Carey et al. 24. Open triangle, mean composition of bile; solid circle, density gradient fraction. Human bile is supersaturated with respect to 1.75% total solids line, whereas rat bile is not supersaturated. Least‐dense fraction (fraction 1) of both human and rat bile contains vesicles and has a composition very different from other density cuts.

Data from Ulloa 130


Figure 1.

Molecular structure of common bile acids showing common steroid ring and side‐chain structure. Hydroxyl group(s) location and orientation are given for each bile acid.



Figure 2.

Crystal structure of bile acids and salts. A: stereoview of the cell of lithocholic acid: α‐axis projection with c‐axis vertical. Oxygen molecules are blackened. B: stereoview of chenodeoxycholic acid; b‐axis projection. C: stereoview of unit cell of deoxycholic acid: H2O, showing channel filled with H2O molecules. Oxygens of H2O molecules are blackened. D: crystal packing of DCA: Rb viewed along b‐axis, showing planar wavy bilayer patterns. Broken lines, hydrogen bonds; large closed circles, Rb+, smaller closed circles, oxygen from H2O; small open circles, oxygen from the bile acid. E: crystal packing of unit cell of Na‐cholate‐monohydrate b‐axis projection. Wavy bilayer pattern is illustrated. Broken lines, hydrogen bonds; larger closed circles, Na+; smaller closed circles, carbon atoms; open circles, oxygen atoms. F: stereoview of bilayer packing of calcium cholate chloride heptahydrate. Closed circles of cholate molecules represent oxygen atoms of hydroxyl and carboxyl groups. Closed circles between cholate molecules represent Ca2+. G: stereoview of bilayer packing of sodium cholate monohydrate. Closed circles of cholate molecules represent oxygen atoms of hydroxyl and carboxyl groups. Closed circles between molecules represent H2O. H: stereoview of bilayer packing of rubidium deoxycholate. Closed circles of deoxycholate molecules represent oxygen atoms of hydroxyl and carboxyl groups. Closed circles between molecules represent H2O.

A from Arora et al. 4; B from Lindley et al. 68; C from Tang et al. 128; D from Coiro et al. 30; E from Cobbledick and Einstein 29; F–H from Hogan et al. 48a)


Figure 3.

Schematic of surface configuration of cholanic acid and hydroxylate derivatives. Maximum area per molecule was taken from first inflection point of compression isotherms; minimum area was taken from collapse point. Pressure at film collapse and state of film are given. Collapse pressure for chenodeoxycholic acid in 3 M NaCl low pH was determined to be ∼27 dyn/cm, similar to that of deoxycholic acid.

Data from Carey 20 and Small 113 and references therein


Figure 4.

Reverse‐phase high‐performance liquid chromatography (HPLC) chromatographs of mixture of bile salts. For a given conjugation state, mobility decreases in the order UDCA > CA > CDCA > DCA. Column used was an Ultrasphere ODS 250 X 4.6 mm, with a mobile phase of 75% MeOH‐25% 0.005 M KH2PO4/H3PO3, pH 5.0. TUDC, tauroursodexoycholate; TC, taurocholate; TCDC, taurochenodeoxycholate; TDC, taurodeoxycholate; GUDC, glycoursodeoxycholate; GC, glycocholate; GCDC, glycochenodeoxycholate; GDC, glycodeoxycholate; UDC, ursodeoxycholate; C, cholate; CDC, chenodeoxycholate; DC, deoxycholate. OD, optical density.

From Armstrong and Carey 3


Figure 5.

Hypothetical titration curve for solutions of free bile salts or for glycine conjugates. W, first equivalence point where titration of bile salt with hydrochloric acid commences. Y, last point where bile salt solution is in thermodynamic equilibrium as a single aqueous phase. T, Tyndall effect noted in this region of titration curve. X, point where precipitation of bile acid crystals commences. X', equilibrium pH at point of bile acid precipitation. Z, second equivalence point where titration of bile salt with hydrochloric acid is complete. TOT, total amount of acid required to complete the titration. HA, amount of acid added from first equivalence point W to point Y, which represents maximum solubility of bile acid (HA) in bile salt solution (A‐).

From Small 113


Figure 6.

Titration curves for cholic acid (CA) in several molecular environments. The pH is changed by adding HCl or NaOH, and chemical shift of carboxyl carbon (C‐24) is monitored by 13C NMR. Aqueous monomeric CA (.2 mM below its CMC and solubility limit). Aqueous micellar CA 116 mM). CA‐sodium taurocholate mixed micelles 1:7 mol/mol). CA‐BSA (bovine serum albumin) complexes 2:1 mol/mol). CA–egg yolk lecithin vesicles 1:20 mol/mol), major peak from CA in outer monolayer of vesicle, minor peak from CA in inner monolayer of vesicle.

Adapted from Cabral et al. 15


Figure 7.

Schematic of structure of bile salt micelles. Molecules of primary micelles associate via hydrophobic regions with aggregation numbers from 2 to 10. At higher concentrations some bile salts form larger aggregates, secondary micelles, stabilized by hydrophobic associations and hydrogen bonding.

From Small 112, printed with permission from Advances in Chemistry Series, copyright 1968, American Chemical Society


Figure 8.

Lecithin–sodium cholate–water diagram. W, H2O; L, lecithin; NaC, sodium cholate. Along side W‐L, numbers indicate H2O wt percentage; along side L‐NaC, numbers indicate lecithin wt percentage; along side NaC‐W, numbers indicate NaC wt percentage. Solid lines, well‐defined boundaries (+L2%); broken lines, less well‐defined frontiers. I, Zone of neat phase. II, Zone of cubic phase. III, Zone of middle phase. IV, Zone of isotropic phase. 1, Zone of separation of neat phase and NaC crystals. 2, Zone of separation of cubic phase and NaC crystals. 3, Zone of separation of middle phase and isotropic solution. 4, Zone of separation of neat phase and cubic phase. 5, Zone of separation of cubic phase and middle phase. 6, Zone of separation of neat phase and middle phase. 7, Zone of separation of neat phase and isotropic phase. 8, Zone of separation of isotropic solution and NaC crystals, a–e, Zones of separation of 3 phases whose compositions for each zone are indicated by apices of the triangles. Points A–H, J, K, M, O–Q are discussed in text. Right side of diagram, molar ratios of lecithin to NaC.



Figure 9.

Lecithin–sodium cholate water diagram at high‐H2O concentrations. NaC, sodium cholate; W, H2O. Left: larger numbers, wt%, smaller numbers, concentrations (mM). Parts of single‐phase zones I, III, and IV are shown, as well as 2‐phase regions 3, 6, and 7. Approximate tie lines in regions 6 and 7 and 3‐phase region are illustrated. Right, Bragg spacings (D) from lamellar and hexagonal lattices. These lattice parameters are given in angstroms along high‐H2O phase boundaries of lamellar phase and hexagonal phase. Dashed line from P (critical micellar concentration of NaC in H2O) to O, separates micelles formed by dilution of hexagonal phase (below) from micelles formed by dissolution of NaC crystals (above).



Figure 10.

Lecithin–sodium cholate system at >90% H2O, illustrating different techniques for preparing mixtures. NaC, sodium cholate; W, H2O. NaC‐W axis: larger numbers, wt% NaC from 0% to 10%; smaller numbers, concentration (mM). Point P, critical micelle concentration of NaC in H2O (.52% or ∼12 mM). Lecithin axis: large numbers, wt% lecithin; small numbers, concentrations (mM). Zone IV has been divided into 2 regions: IVa and IVb. Line separating 2 regions is a straight line running from point P to point O (see Fig. 9 and indicates a change in micellar structure. Narrow 2‐phase region 3 and larger 2‐phase region 7 are spearated by 3‐phase region a. Heavy lines, various methods of preparing mixtures. Any mixture may be made up by coprecipitation method (e.g., points 1 on 10% solid line and 5% solid line; TS, total solutes). A mixture made up by coprecipitation may then be diluted with H2O or buffer and will follow line 2. If a coprecipitated mixture is diluted with a specific concentration of bile salt, e.g., 2%, it will follow line 3. A coprecipitated mixture such as that plotted on 5% line at 40 mM lecithin and 45 mM NaC may be placed in a dialysis bag and dialyzed against H2O, buffer, or a bile salt solution. Because lecithin does not escape from the dialysis bag and its concentration remains unchanged, the composition will move toward W‐lecithin side along a constant 40‐mM line, line 4. Concentration of dialysate (not shown) will be found along W‐NaC axis. Finally, any concentration of lecithin, eg., a 6% suspension (either coarse liposomes or unilamellar vesicles), may be diluted with a specific concentration of bile salt, for instance at 10%, as shown on line 5. Compositions will fall on line 6, depending on amount of 10% bile salt solution added.



Figure 11.

A: mean hydrodynamic radii () of sodium taurocholate‐lecithin solutions (L/NaTC) at various total lipid concentrations (.625%–10% at 24°C in 0.15 M NaCl). diverges as lecithinbile salt ratio approaches micellar‐phase limits, and divergence limits are very concentration dependent. Dashed line, radius of simple discoid micelle. Phase limits shown at bottom of each panel represent appropriate lecithin‐bile salt ratio for total lipid concentrations of a, 0.0625%; b, 1.25%; c, 2.5%; d, 5.0%; and e, 10.0%. values indicated at tops of certain curves represent size of bilayer vesicles formed in supersaturated systems. B: at 3 L‐NaTC ratios as a function of the total wt% of lipids. As the total percentage of lipids increases, decreases markedly and approaches the dimensions of the simple discoidal micelle (arrows at right) at high lipid concentrations.

A reprinted with permission from Mazer et al. 70. Copyright 1980 American Chemical Society


Figure 12.

Lecithin‐sodium taurocholate (NaTC) system at high H2O (W) content. A: divergence limit (see Fig. 11A) and coexistence boundary plotted on rectangular coordinates. B‐C: system plotted on triangular coordinates (note difference in scale). Dashed line, phase boundary between micellar phase and zone 3. Phase boundary is virtually same as divergence limit line. Point P (critical micellar concentration of NaTC in H2O) has a value of 3.2 mM; 2‐phase regions 3 and 7 are shown. Tie lines in region 7 and the 3‐phase region a are also shown.

A from Mazer et al. 70; B‐C, coexistence and divergence boundaries from Mazer et al. 70


Figure 13.

A: mean hydrodynamic radius . B: polydispersity index V of a mixed micellar stock solution sodium glycocholatelecithin [lecithin/sodium glycocholate = 0.85; total concentration (Ctot) = 5 g/dl; temperature = 20°C] as a function of dilution. Dilution factor of 1 corresponds to stock solution Ctot, and dilution factor of n corresponds to Ctot divided by n.

From Schurtenberger et al. 108. Reprinted with permission from Journal of Physical Chemistry, copyright 1985, American Chemical Society


Figure 14.

Lecithin‐sodium glycocholate‐water system in dilute region of zone 7. L, lecithin; NaGC, sodium glycocholate; W, water. Closed circles, composition of fluid within dialysis bag; open circles, corresponding compositions (connected by a line) of dialysate fluid. Lines connect aqueous phase (intermicellar concentration or dialysate) with mixture composition (closed circles) for a series of vesicles of different sizes [ (Å)]. Extensions of these lines intersect L‐NaGC border (as shown by closed circles on right) and give ratio of NaGC bound to lecithin vesicle. Rh, mean hydrodynamic radii.



Figure 15.

Rates of dissolution of multilamellar liposomes by bile salts (BS) of different concentration. Lecithin concentrations of 1.5 mM were diluted with an equal volume of bile salts at concentrations on horizontal axis. A: time for half dissolution (T1/2 is plotted as a function of bile salt concentration; B: complex rate constant k1 91 for the process is plotted. Order of dissolution rates, chenodeoxycholate (CDC) > deoxycholate (DC) > cholate (C) > ursodeoxycholate (UDC).



Figure 16.

Sodium taurocholate‐lecithin‐cholesterol phase diagram. Chol, cholesterol; NaTC, sodium taurocholate; PL, phospholipid. At 10% total lipid, micellar phase is bounded by solid line. Dotted lines, micellar boundaries with decreasing concentrations of total lipids. Liquid crystal phase is a phospholipid phase with varying amounts of cholesterol and small amounts of bile salts. Crystalline phase is cholesterol monohydrate. We suggest that a 3‐phase region cannot directly abut on a 1‐phase region, except at a point where the 2‐phase region on the right extends to point B. The 3‐phase region consists of 1) cholesterol crystals, 2) liquid crystals of phospholipid, cholesterol, and bile salt, and 3) a micellar solution of composition B.

Adapted from Carey et al. 24


Figure 17.

Phase diagrams of aqueous taurine‐conjugated ursodeoxycholic acid (TUDC)‐, cholic acid (TC)‐, or chenodeoxycholic acid (TCDC)‐lecithin‐cholesterol monohydrate at a total lipid concentration of 10 g/100 ml (.20 M Na+, pH 7.4, 37°C). Axes are expressed in mol%. Although phase diagrams show 3‐phase region abutting 1‐phase region, there must be a 2‐phase region, however small, separating them.

From Salvioli et al. 101


Figure 18.

Partial phase diagram for aqueous system sodium taurocholate‐lecithin‐cholesterol 20 g/100 ml, 0.15 M NaCl, 24°C). Shaded area, metastable region; open circles, maximum equilibrium cholesterol solubility; closed circles, metastable‐labile limit.

From Carey et al. 24


Figure 19.

Behavior of hypothetical biles of compositions a‐f as they transform from dilute hepatic bile to concentrated gallbladder bile. Total lipids for hepatic bile are ∼1.5% and for gallbladder bile 10%. Phase boundaries are from Carey et al. 24. BS, bile salt. Point B is the same as that discussed in Fig. 16. For further explanation, see text.



Figure 20.

Electron micrograph of a bile canaliculus of a bile salt‐depleted rat liver that shows the presence of vesicular material within the lumen (left) (X 32,800). Higher magnification (inset) illustrates more clearly unilamellar structure of vesicle and can be compared with structure of canalicular membrane. Mean diameter of these vesicles is 56 nm 30–85 nm). Similar vesicles are also apparent in fresh bile specimen of same animal (right). Bars, 0.25 μm.

From Ulloa et al. 130, © by Am. Assoc. of the Study of Liver Diseases, 1987


Figure 21.

Distribution and composition of biliary lipids in bile fractionated on a continuous metrizamide density gradient. Left, T‐tube human hepatic bile; right, rat hepatic bile. Histograms are mean +L SD of percentage of each lipid in total sample of 7 human biles and 4 rat biles. Total lipid composition of rat biles was 2.4 g/dl; total composition of human T‐tube biles was 1.7 g/dl. BS, bile salt; C, cholesterol; PL, phospholipid. Compositions of each of the density cuts are plotted below on triangular coordinates with appropriate line as determined by Carey et al. 24. Open triangle, mean composition of bile; solid circle, density gradient fraction. Human bile is supersaturated with respect to 1.75% total solids line, whereas rat bile is not supersaturated. Least‐dense fraction (fraction 1) of both human and rat bile contains vesicles and has a composition very different from other density cuts.

Data from Ulloa 130
References
 1. Admirand, W. H., and D. M. Small. The physico‐chemical basis of cholesterol gallstone formation in man. J. Clin. Invest. 47: 1045–1052, 1968.
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Article Title: Physical Chemistry of Bile
 

How to Cite

Donna J. Cabral, Donald M. Small. Physical Chemistry of Bile. Compr Physiol 2011, Supplement 18: Handbook of Physiology, The Gastrointestinal System, Salivary, Gastric, Pancreatic, and Hepatobiliary Secretion: 621-662. First published in print 1989. doi: 10.1002/cphy.cp060331