Fracture-toughening mechanisms responsible for differences in work to fracture of hydrated and dehydrated dentine

This study investigates the nature of deformation and differences in the mechanisms of fracture and properties of dentine where there has been a loss of moisture, as may occur with removal of the pulp in the endodontic treatment of teeth. Controlled fracture toughness testing was conducted on bovine teeth to determine the influence of hydration on the work of fracture of dentine. Significant differences (p<0.01) were observed between the fracture toughness of hydrated (554+/-27.7J/m2) and dehydrated (113+/-17.8J/m2) dentine. Observations of the crack tip region during crack extension revealed extensive ligament formation occurred behind the crack tip. These ligaments provide considerable stability to the crack by significantly increasing the work of fracture, thereby acting as a fracture-toughening mechanism. Micro-cracking, reported as a fracture-toughening mechanism in bone, is also clearly seen. A zone of in-elastic deformation may occur as hydrated specimens revealed upon crack extension, a region about the tip that appeared to suck water into the structure and to exude water behind the crack tip. In dehydrated dentine, no in-elastic zone was observed. Micro-cracking is present though the cracks are smaller, straighter and with less opening than hydrated dentine. Only limited ligament formation just behind the crack tip was observed. These differences resulted in a significantly lower work of fracture with unstable brittle fracture characteristics. Based on these results, several fracture-toughening mechanisms were identified in dentine, with micro-cracking not considered the most important. These findings may be relevant for bone, a similar mineralised hydrated tissue.     

Fracture toughness of manatee rib and bovine femur using a chevron-notched beam test

Bone is an anisotropic material with a hierarchical structure consisting of organic matrix, minerals and water. Fracture toughness (K(C)) has been shown to be a good index to assess the mechanical performance of bone. A chevron-notched (CN) beam test, a standard fracture mechanics test successfully applied to many other materials, was used to determine the transverse-direction fracture toughness in manatee rib and bovine femur cortical bone. Although human and bovine bone has been well studied, there is virtually no information on the toughness of manatee rib bone. As a biological material, manatee rib is interesting for study in that it is a highly mineralized bone. Three major advantages of the CN specimen test are: (1) it is easier to reach plane strain condition; (2) there is no fatigue-precracking needed; and (3) it is relatively easy to produce stable crack propagation before catastrophic fracture. The fracture toughness values of manatee rib and bovine femur were measured to be 4.5 +/- 0.5 MPa m(1/2) and 5.8 +/- 0.5 MPa m(1/2), respectively. Based on the microstructures shown in SEM images, two features that contributed to the greater fracture toughness of bovine femur were identified as greater osteon density and lesser porosity.     

Effect of specimen thickness on fracture toughness of bovine patellar cartilage

Fracture toughness and crack tip opening angle were measured for bovine patellar cartilage using modified single-edged notch specimens of two thicknesses. There was no difference in fracture toughness between thin (0.7 mm) versus relatively thick (2.7 mm) specimens, but the crack tip opening angle at initiation of crack propagation was larger for the thin specimens (106 deg) than for the thick specimens (70 deg). Fracture toughness of the bovine patellar cartilage (1.03 kJ/m2) was not statistically different than that reported previously for canine patellar cartilage (1.07 kJ/m2) employing the same methods. Large variation in measurements for both bovine and canine cartilage are in part attributable to variation between individual animals, and are consistent with variation in other mechanical property measurements for articular cartilage. The observed reduction in crack tip opening angle with increased specimen thickness is consistent with behavior of some engineering materials, and demonstrates that specimen thickness influences fracture behavior for bovine patellar cartilage.     

Effect of temperature on the fracture toughness of compact bone

Bone is a composite composed mainly of organics, minerals, and water. Many researchers have studied effects such as crack velocity, density, orientation, storage media, porosity, and age on the fracture toughness (K(C), also called critical stress intensity factor) of compact bone. Most of these studies were conducted at room temperature. Considering that the body temperature of animals is greater than room temperature, and that bone has a large volumetric percentage of organics and water (generally, 55-65%), it is hypothesized that temperature has a significant effect on the fracture toughness of compact bone. Single-edge V-notched (SEVN) specimens were prepared to measure the fracture toughness of bovine femur and manatee rib in water at 0, 10, 23, 37, and 50 degrees C in four-point flexure. The fracture toughness values of bovine femur and manatee rib were found to decrease from 7.0 to 4.3MPam(1/2) and from 5.5 to 4.0MPam(1/2), respectively, as temperature increased over a temperature range of 50 degrees C. The results support the hypothesis that temperature has a significant effect on the fracture toughness of compact bone. Therefore, we suggest that study on fracture toughness of bone should be done at physiologically relevant temperatures.     

The effect of moisture absorption on the fatigue crack propagation resistance of acrylic bone cement.

In vivo, bone cement is subject to cyclic loading in a fluid environment. However, little is known about the effect of moisture absorption on the fatigue crack propagation resistance of bone cement. The effect of moisture absorption at 37 degrees C on the fatigue crack propagation resistance of a common bone cement (Endurance, DePuy, Orthopaedics, Inc.) was examined. Preliminary fracture toughness tests were conducted on disk-shaped, vacuum-mixed cement specimens (compact tension type) that were cyclically pre-cracked. Plain-strain fracture toughness K(IC) (MPa square root(m)) was determined. To study the effect of moisture absorption four treatment groups, with different soaking periods in Ringer's at 37 degrees C, of Endurance cement were tested. The specimens weights prior to and following soaking showed a significant increase in mean weight for specimens soaked for 8 and 12 weeks. Linear regression analysis of log(da/dN) vs. log (deltaK) was conducted on the combined data in each fatigue test group. Soaking bone cement in Ringer's at 37 degrees C for 8 and 12 weeks lead to an improvement in fatigue crack propagation resistance, that may be related to water sorption that increases polymer chain mobility, with enhanced crack tip blunting. It may be more physiologically relevant to conduct in vitro studies of fatigue and fracture toughness of bone cements following storage in a fluid environment.     

Application of fracture mechanics to failure in manatee rib bone

BACKGROUND: The Florida manatee (Trichechus manatus latirostris) is listed as endangered by the U.S. Department of the Interior. Manatee ribs have different microstructure from the compact bone of other mammals. Biomechanical properties of the manatee ribs need to be better understood. Fracture toughness (K(C)) has been shown to be a good index to assess the mechanical performance of bone. Quantitative fractography can be used in concert with fracture mechanics equations to identify fracture initiating defects/cracks and to calculate the fracture toughness of bone materials. METHOD OF APPROACH: Fractography is a standard technique for analyzing fracture behavior of brittle and quasi-brittle materials. Manatee ribs are highly mineralized and fracture in a manner similar to quasi-brittle materials. Therefore, quantitative fractography was applied to determine the fracture toughness of manatee ribs. RESULTS: Average fracture toughness values of small flexure specimens from six different sizes of manatees ranged from 1.3 to 2.6 MPa(m)(12). Scanning electron microscope (SEM) images show most of the fracture origins were at openings for blood vessels and interlayer spaces. CONCLUSIONS: Quantitative fractography and fracture mechanics can be combined to estimate the fracture toughness of the material in manatee rib bone. Fracture toughness of subadult and calf manatees appears to increase as the size of the manatee increases. Average fracture toughness of the manatee rib bone materials is less than the transverse fracture toughness of human and bovine tibia and femur.     

Evaluation of fracture toughness of human dentin using elastic-plastic fracture mechanics

Dentin, the mineralized tissue forming the bulk of the tooth, lies between the enamel and the pulp chamber. It is a rich source of inspiration for designing novel synthetic materials due to its unique microstructure. Most of the previous studies investigating the fracture toughness of dentin have used linear-elastic fracture mechanics (LEFM) that ignores plastic deformation and could underestimate the toughness of dentin. With the presence of collagen (approximately 30% by volume) aiding the toughening mechanisms in dentin, we hypothesize that there is a significant difference between the fracture toughness estimated using LEFM (Kc) and elastic-plastic fracture mechanics (EPFM) (KJc). Single-edge notched beam specimens with in-plane (n=10) and anti-plane (n=10) parallel fractures were prepared following ASTM standard E1820 and tested in three-point flexure. KJc of the in-plane parallel and anti-plane parallel specimens were found to be 3.1 and 3.4 MPa m 1/2 and Kc were 2.4 and 2.5 MPa m 1/2, respectively. The fracture toughness estimated based on KJc is significantly greater than that estimated based on Kc (32.5% on average; p<0.001). In addition, KJc of anti-plane parallel specimens is significantly greater than that of in-plane parallel specimens. We suggest that, in order to critically evaluate the fracture toughness of human dentin, EPFM should be employed.     

Contribution, development and morphology of microcracking in cortical bone during crack propagation

A fracture mechanics study of cortical bone is presented to investigate the contribution, development morphology of microcracking in cortical bone during crack propagation. Post-hoc analyses of microcrack orientation, crack propagation velocity and fracture surface roughness were conducted on previously tested human and bovine bone compact tension specimens. It was found that, consistent with its higher toughness, bovine bone formed significantly more longitudinal, transverse and inclined microcracks than human bone. However, in human bone more of the microcracks that formed were longitudinal than transverse or inclined, a feature that would optimise bone's toughness. Crack propagation velocity in human and bovine bone displayed the same characteristic pattern with crack extension, where an increase in velocity is followed by a consequent decrease and vice versa. On the basis of this pattern, a model or crack propagation has been proposed. It provides a detailed account of mocrocrack formation and contribution towards the propagation of a fracture crack. Analyses of fracture surfaces indicated that, consistent with its higher toughness, bovine bone displays a rougher surface than human bone but they both have the same basic fractured element, i.e. a mineralised collagen fibril.     

Lateral diffusion rates of lipid,water, and a hydrophobic drug in a multilamellar liposome

The lateral diffusion constants of 1-palmitoyl-2-oleoyl-sn-glycero-3 phosphocholine (POPC), water, and ibuprofen were measured in multilamellar liposomes using pulsed field gradient magic-angle spinning (PFG-MAS) (1)H NMR. The analysis of diffusion data obtained in powder samples and a method for liposome curvature correction are presented. At 322 K POPC has a diffusion constant of (8.6 +/- 0.2) x 10(-12) m(2)/s when dehydrated (8.2 waters/lipid) and (1.9 +/- 0.1) x 10(-11) m(2)/s in excess water. The diffusion constant of water in dehydrated POPC was found to be (4.7 +/- 0.1) x 10(-10) m(2)/s. The radius of curvature is 21 +/- 2 microm for the dehydrated sample and 4.5 +/- 0.5 microm for POPC sample containing excess water. The activation energies of diffusion are 40.6 +/- 0.4 kJ/mole for dehydrated POPC, 30.7 +/- 0.9 kJ/mole for POPC with excess water, and 28.6 +/- 1.5 kJ/mole for water in dehydrated POPC. The diffusion constants and activation energies for a sample of POPC/ibuprofen/water (1:0.56:15) were also measured. The ibuprofen, which locates in the lipid-water interface, diffuses faster than POPC but has a slightly higher activation energy of lateral diffusion. Within certain restrictions, PFG-MAS NMR provides a useful method for characterizing membrane organization and mobility.     

The influence of water removal on the strength and toughness of cortical bone

Although the effects of dehydration on the mechanical behavior of cortical bone are known, the underlying mechanisms for such effects are not clear. We hypothesize that the interactions of water with the collagen and mineral phases each have a unique influence on mechanical behavior. To study this, strength, toughness, and stiffness were measured with three-point bend specimens made from the mid-diaphysis of human cadaveric femurs and divided into six test groups: control (hydrated), drying in a vacuum oven at room temperature (21 degrees C) for 30 min and at 21, 50, 70, or 110 degrees C for 4 h. The experimental data indicated that water loss significantly increased with each increase in drying condition. Bone strength increased with a 5% loss of water by weight, which was caused by drying at 21 degrees C for 4 h. With water loss exceeding 9%, caused by higher drying temperatures (> or =70 degrees C), strength actually decreased. Drying at 21 degrees C (irrespective of time in vacuum) significantly decreased bone toughness through a loss of plasticity. However, drying at 70 degrees C and above caused toughness to decrease through decreases in strength and fracture strain. Stiffness linearly increased with an increase in water loss. From an energy perspective, the water-mineral interaction is removed at higher temperatures than the water-collagen interaction. Therefore, we speculate that loss of water in the collagen phase decreases the toughness of bone, whereas loss of water associated with the mineral phase decreases both bone strength and toughness.     

Effect of specimen size on work-of-fracture measurements

It has been suggested that work-of-fracture, which quantifies the ability of a material to resist fracture, is dependent on specimen size. This experiment compared work-of-fracture, calculated as energy per unit area, for different specimen sizes of Plexiglas, bovine tibial bone and aluminum. Three different geometrically similar cross sections were tested for each material for a total of 54 specimens. Work-of-fracture was measured by loading a notched beam (triangular cross section) in three-point bending at a constant deformation rate. The energy necessary to cause fracture was measured from a load-deformation curve. Specimen fracture area was determined using macrophotography. Atomic absorption spectrophotometry was used to determine weight percent calcium of bone specimens and quantitative light microscopy was used to determine fractional void area. Analysis of variance showed no effect of specimen size on work-of-fracture for aluminum or Plexiglas specimens (p greater than 0.05). A significant difference was found, however, between the large (area = 11.7 +/- 1.9 mm2) and small (area = 3.48 +/- 0.68 mm2) bone specimens and between the medium (area = 5.89 +/- 0.69 mm2) and small (area = 3.48 +/- 0.68 mm2) bone specimens. No correlation was found between work-of-fracture and either calcium content (r2 = 0.128) or fractional void area (r2 = 0.0713). The mean work-of-fracture values found are as follows: aluminum, 59.8 +/- 13.7 kJ m-2; Plexiglas, 0.620 +/- 0.074 kJ m-2; bone (area 5.89 +/- 0.69 mm2-11.7 +/- 1.9 mm2), 9.72 +/- 1.93 kJ m-2 and bone (area 3.48 +/- 0.68 mm2), 5.48 +/- 1.79 kJ m-2.     

Fracture toughness and fatigue crack propagation rate of short fiber reinforced epoxy composites for analogue cortical bone

Third-generation mechanical analogue bone models and synthetic analogue cortical bone materials manufactured by Pacific Research Laboratories, Inc. (PRL) are popular tools for use in mechanical testing of various orthopedic implants and biomaterials. A major issue with these models is that the current third-generation epoxy-short fiberglass based composite used as the cortical bone substitute is prone to crack formation and failure in fatigue or repeated quasistatic loading of the model. The purpose of the present study was to compare the tensile and fracture mechanics properties of the current baseline (established PRL "third-generation" E-glass-fiber-epoxy) composite analogue for cortical bone to a new composite material formulation proposed for use as an enhanced fourth-generation cortical bone analogue material. Standard tensile, plane strain fracture toughness, and fatigue crack propagation rate tests were performed on both the third- and fourth-generation composite material formulations using standard ASTM test techniques. Injection molding techniques were used to create random fiber orientation in all test specimens. Standard dog-bone style tensile specimens were tested to obtain ultimate tensile strength and stiffness. Compact tension fracture toughness specimens were utilized to determine plane strain fracture toughness values. Reduced thickness compact tension specimens were also used to determine fatigue crack propagation rate behavior for the two material groups. Literature values for the same parameters for human cortical bone were compared to results from the third- and fourth-generation cortical analogue bone materials. Tensile properties of the fourth-generation material were closer to that of average human cortical bone than the third-generation material. Fracture toughness was significantly increased by 48% in the fourth-generation composite as compared to the third-generation analogue bone. The threshold stress intensity to propagate the crack was much higher for the fourth-generation material than for the third-generation composite. Even at the higher stress intensity threshold, the fatigue crack propagation rate was significantly decreased in the fourth-generation composite compared to the third-generation composite. These results indicate that the bone analogue models made from the fourth-generation analogue cortical bone material may exhibit better performance in fracture and longer fatigue lives than similar models made of third-generation analogue cortical bone material. Further fatigue testing of the new composite material in clinically relevant use of bone models is still required for verification of these results. Biomechanical test models using the superior fourth-generation cortical analogue material are currently in development.     

Effect of groove on bone fracture toughness

When testing for the effects of bone orientation on mode I fracture toughness, compact tension specimens are grooved with a V-notch to provide a crack guide. The effect of grooving on the expressions for the critical stress intensity factor (K_c) and the critical strain energy release rate (G_c) for mode I fracture toughness was investigated. Experiments were performed using grooved and ungrooved bovine compact tension specimens. The results indicate that the standard expression used to determine K_c for a compact tension specimen requires modification. The thickness (B) must be modified to account for the thickness between the grooves (B_n). The thickness used in the standard expression is replaced by an effective thickness written as (BB_n)^0.5. It was also found that the thickness between the grooves should be used in the standard formula for G_c.     

Direct observation of the enthalpy change accompanying the native to molten-globule transition of cytochrome c by using isothermal acid-titration calorimetry

The enthalpy change accompanying the reversible acid-induced transition from the native (N) to the molten-globule (MG) state of bovine cytochrome c was directly evaluated by isothermal acid-titration calorimetry (IATC), a new method for evaluating the pH dependence of protein enthalpy. The enthalpy change was 30 kJ/mol at 30 degrees C, pH 3.54, with 500 mM KCl. The results of the global analysis of the temperature dependence of the excess enthalpy from 20 to 35 degrees C demonstrated that the N to MG transition is a two-state transition with a small heat capacity change of 1.1 kJ K(-1) mol(-1). The present findings were also indicative of the pH dependence of the enthalpy and the heat capacity of the MG state, -13 kJ mol(-1) pH(-1) and -1.0 kJ K(-1) mol(-1) pH(-1), respectively, at 30 degrees C within a pH range from 2 to 3.     

Thermoregulatory responses to exercise in dehydrated dogs

Measurements of rectal temperature (Tre), water lost by evaporation (Eresp) and drooling, cardiac output (CO), and common carotid blood flow (CCBF) were made in dogs (mean hydrated wt 31.0 +/- 1.5 kg) running for 1 h on a level treadmill at 7.5 km/h at an ambient temperature of 25 degrees C. Each animal was studied when it was hydrated ad libitum and when it had been dehydrated by removal of drinking water until 9-10% of the initial body weight had been lost. Dehydrated exercising animals had significantly higher Tre and lower rates of Eresp, CO, and CCBF. Tre and Eresp were measured in seven animals. Average Tre during running was 39.11 +/- 0.10 degrees C in hydrated and 39.80 +/- 0.25 degrees C in dehydrated animals (P less than 0.01). Average Eresp during running was 3.9 +/- 0.3 g/min in hydrated animals and 2.3 +/- 0.3 g/min in dehydrated animals (P less than 0.01). Average CO during exercise, measured in five animals, was 11.1 +/- 0.7 1/min in the hydrated state and 8.6 +/- 0.5 1/min in the dehydrated state (P less than 0.01). Unilateral CCBF during exercise, measured in four animals, was 602 +/- 40 ml/min in the hydrated state and 418 +/- 22 ml/min in the dehydrated state (P less than 0.01). Water lost by drooling in seven exercising animals was 41.5 +/- 11 g/h when they were hydrated and 0.6 +/- 0.4 g/h when they were dehydrated. It is concluded that dehydrated dogs doing mild exercise can save water by reducing Eresp and regulating body temperature above hydrated levels.(ABSTRACT TRUNCATED AT 250 WORDS)     

Specimen size effect in the volumetric shrinkage of cancellous bone measured at two levels of dehydration

Water is commonly removed from bone to study its effect on mechanical behaviour; however, dehydration also alters the bone structure. To make matters worse, measuring structural changes in cancellous bone is complicated by a number of factors. Therefore, the goals of this study were to address these issues by (1) comparing Archimedes' method and a helium pycnometer as methods for measuring cancellous bone volume; (2) measuring the apparent dimensional and volumetric tissue shrinkage of cancellous bone at two levels of dehydration; and, (3) identifying whether a size effect exists in cancellous bone shrinkage. Cylindrical specimens (3, 5 and 8.3 mm diameters) of cancellous bone were taken from the distal bovine femur. The apparent dimensions of each cylindrical specimen were measured in a fully hydrated state (HYD), after drying at room temperature (AIR), and after oven drying at 105 degrees C (OVEN). Tissue volume measurements for those three hydration states were obtained using both a helium pycnometer and Archimedes' method. Aluminium foams, which mimic the cancellous structure, were used as controls. The results suggest that the helium pycnometer and Archimedes' method yield identical results in the HYD and AIR states, but that Archimedes' method under-predicts the nominal OVEN volume by incorporating the collagen-apatite porosity. A distinct size effect on volumetric shrinkage is observed (p<0.025) using the pycnometer in both AIR and OVEN states. Apparent dimensional shrinkage (2% and 7%) at the two dehydration levels is much smaller than the measured volumetric tissue shrinkage (16% and 29%), which results in a reduced dehydrated bone volume fraction.     

Phase behavior and glass transition of 1,2-dioleoylphosphatidylethanolamine (DOPE) dehydrated in the presence of sucrose

The effect of sucrose on the phase behavior of 1,2-dioleoylphosphatidylethanolamine (DOPE) as a function of hydration was studied using differential scanning calorimetry and X-ray diffraction. DOPE/sucrose/water dispersions were dehydrated at osmotic pressures (Pi) ranging from 2 to 300 MPa at 30 degrees C and 0 degrees C. The hexagonal II-to-lamellar gel (H(II)-->L(beta)) thermotropic phase transition was observed during cooling in mixtures dehydrated at Pior=57 MPa, the H(II)-->L(beta) thermotropic phase transition was precluded when sucrose entered the rigid glassy state while the lipid was in the H(II) phase. Sucrose also hindered the H(II)-to-lamellar crystalline (L(c)), and H(II)-to-inverted ribbon (P(delta)) lyotropic phase transitions, which occurred in pure DOPE. Although the L(c) phase was observed in dehydrated 2:1 (mole ratio) DOPE/sucrose mixtures, it did not form in mixtures with higher sucrose contents (1:1 and 1:2 mixtures). The impact of sucrose on formation of the ordered phases (i.e., the L(c), L(beta), and P(delta) phases) of DOPE was explained as a trapping of DOPE in a metastable H(II) phase due to increased viscosity of the sucrose matrix. In addition, a glass transition of DOPE in the H(II) phase was observed, which we believe is the first report of a glass transition in phospholipids.     

Rising crack-growth-resistance behavior in cortical bone: implications for toughness measurements

Fracture mechanics studies have characterized bone's resistance to fracture in terms of critical stress intensity factor and critical strain energy release rate measured at the onset of a fracture crack. This approach, although useful, provide a limited insight into fracture behavior of bone because, unlike classical brittle materials, bone is a microcracking solid that derives its resistance to fracture during the process of crack propagation from microfracture mechanisms occurring behind the advancing crack front. To address this shortfall, a crack propagation-based approach to measure bone toughness is described here and compared with crack initiation approach. Post hoc analyses of data from previously tested bovine and antler cortical bone compact specimens demonstrates that, in contrast to crack initiation approach, the crack propagation approach successfully identifies the superior toughness properties of red deer's antler cortical bone. Propagation-based slope of crack growth resistance curve is, therefore, a more useful parameter to evaluate cortical bone fracture toughness.     

Stability and binding properties of wild-type and c17s mutated human sterol carrier protein 2.

The temperature- and solvent-induced denaturation of both the SCP2 wild-type and the mutated protein c71s were studied by CD measurements at 222 nm. The temperature-induced transition curves were deconvoluted according to a two-state mechanism resulting in a transition temperature of 70.5 degrees C and 59.9 degrees C for the wild-type and the c71s, respectively, with corresponding values of the van't Hoff enthalpies of 183 and 164 kJ/mol. Stability parameters characterizing the guanidine hydrochloride denaturation curves were also calculated on the basis of a two-state transition. The transitions of the wild-type occurs at 0.82 M GdnHCl and that of the c71s mutant at 0.55 M GdnHCl. These differences in the half denaturation concentration of GdnHCl reflect already the significant stability differences between the two proteins. A quantitative measure are the Gibbs energies DeltaG(0)(D)(buffer) at 25 degrees C of 15.5 kJ/mol for the wild-type and 8.0 kJ/mol for the mutant. We characterized also the alkyl chain binding properties of the two proteins by measuring the interaction parameters for the complex formation with 1-O-Decanyl-beta-D-glucoside using isothermal titration microcalorimetry. The dissociation constants, K(d), for wild-type SCP2 are 335 microM at 25 degrees C and 1.3 mM at 35 degrees C. The corresponding binding enthalpies, DeltaH(b), are -21. 5 kJ/mol at 25 degrees C and 72.2 kJ/mol at 35 degrees C. The parameters for the c71s mutant at 25 degrees C are K(d)=413 microM and DeltaH(b)=16.6 kJ/mol. These results suggest that both SCP2 wild-type and the c71s mutant bind the hydrophobic compound with moderate affinity.     

An investigation of the equilibria between aqueous ribose, ribulose, and arabinose

The thermodynamics of the equilibria between aqueous ribose, ribulose, and arabinose were investigated using high-pressure liquid chromatography and microcalorimetry. The reactions were carried out in aqueous phosphate buffer over the pH range 6.8-7.4 and over the temperature range 313.15-343.75 K using solubilized glucose isomerase with either Mg(NO3)2 or MgSO4 as cofactors. The equilibrium constants (K) and the standard state Gibbs energy (delta G degrees) and enthalpy (delta H degrees) changes at 298.15 K for the three equilibria investigated were found to be: ribose(aq) = ribulose(aq) K = 0.317, delta G degrees = 2.85 +/- 0.14 kJ mol-1, delta H degrees = 11.0 +/- 1.5 kJ mol-1; ribose(aq) = arabinose(aq) K = 4.00, delta G degrees = -3.44 +/- 0.30 kJ mol-1, delta H degrees = -9.8 +/- 3.0 kJ mol-1; ribulose(aq) = arabinose(aq) K = 12.6, delta G degrees = -6.29 +/- 0.34 kJ mol-1, delta H degrees = -20.75 +/- 3.4 kJ mol-1. Information on rates of the above reactions was also obtained. The temperature dependencies of the equilibrium constants are conveniently expressed as R in K = -delta G degrees 298.15/298.15 + delta H degrees 298.15[(1/298.15)-(1/T)] where R is the gas constant (8.31441 J mol-1 K-1) and T the thermodynamic temperature.