Thermal stability of whey proteins studied by differential scanning calorimetry

The thermal stability of the bovine whey proteins.; β-lactoglobulin (β-1g), α-lactalbumin (α-1a) and serum albumin (BSA) was studied individually and in mixtures in the temperature range 25–140°C by differential scanning calorimetry. The thermal denaturation temperature (T_D) and the transition enthalpies (ΔH_app) were determined at different pH-values (3.0–10.0) in simulated milk ultrafil-trate (SMUF).β-Lg was, except at pH 9.0 and 10.0, the most thermostable protein at all pH-values. At acidic pH-values BSA was the least thermostable. At alkaline pH-values, however, α-la had lower thermal stability than BSA. α-La exhibited double peak behaviour at acidic pH-values and ΔH_app was dependent on Ca-content. Mixtures of the proteins were studied at pH 4.0, 5.0 and 6.6. In general, when mixed, the proteins seemed to denaturate independently of each other.     

Use of cyclic differential scanning calorimetry

Formation of an activated cellulose (Cellulose) species $$CELLULOSE\xrightarrow[{air}]{{heat}}CELLULOSE*$$ is the designated first stage of cellulose degradation in air [1]. Little is known about either the process or the nature of CELLULOSE^*. The transition, designatedT ₂, is observed as an exotherm around 300°C as the sample temperature is raised. No corresponding endotherm is observed on cooling. The process is therefore not reversible but is repeatable as subsequent reheating results in the exotherm being observed again. The exotherm is also found to be oxygen dependant. The effect of all the flame retardant treatments studied was to reduceT ₂ compared to the value for the untreated cotton.     

Quantitative aspects of differential scanning calorimetry

The quantitative performance of differential scanning calorimeters is reviewed. Temperature calibration is discussed in terms of an isothermal correction plus a contribution from thermal lag, this can be derived from individual curves and is valid in both, heating and cooling. It is emphasised that baselines that are drawn to thermal events, such as melting and transition phenomena, must have thermodynamic significance and a general procedure is suggested. When this is used, a power compensation calorimeter calibrated for heat-capacity work can reproduce heats of fusion and transition for a diverse range of materials to better than 1%.     

Thermal optical analysis and differential scanning calorimetry of chemically crosslinked polyethylene

Differential scanning calorimetry (DSC) and thermal optical analysis (TOA) were applied as part of a series of characterization methods to solid chemically crosslinked polyethylene insulation. The DSC was used to study the melting and recrystallization. The ΔH of fusion has a lower value than expected. The TOA shows premelting behavior and is more sensitive to crystalline charges, on cooling, than DSC.     

Thermal analysis of the system KCL-LiCI by differential scanning calorimetry

Thermal analysis of the binary system KCl-LiCl in the composition range 0.368–0.812 mol fraction of LiCl was studied by differential scanning calorimetry (DSC). On the basis of the DSC curves, the experimental data for the phase-diagram, the latent heat of fusion, and the average specific heat in the liquid and solid states are presented as a function of the composition of the mixture. The experimental results compared with literature data. The following empirical correlation between the heat of fusion (H) and of compositions of the mixture in mol fraction of LiCl (x) was obtained: ·GH=26.95–50.20x+43.06x² with a minimum value of 11.8 kJ(g mol)^–1 at the eutectic point of 0.587 mol fraction of LiCl at 354.4°C. These results are required as basic data to develop thermal energy storage materials, based on the phase change of a molten mixture of KCl-LiCI.     

Precise determination of melting and boiling points by differential thermal analysis and differential scanning calorimetry

The theory, operation and instrumentation of differential thermal analysis (DTA) and differential scanning calorimetry (DSC) are sufficiently well developed to determine melting and boiling points with a high degree of certainty and reproducibility. However, certain precautions must be taken if data of maximum value are to be obtained. Sampling techniques, encapsulation, instrumental parameters and theoretical considerations will be treated in detail.In addition to the very small amount of material required for a melting or boiling point determination, DTA and DSC have other advantages. If certain precautions are observed it is possible to use several equations from classical thermodynamics to obtain absolute purity. A complete Cox Chart of a pure liquid can be obtained and the heat of vaporization determined in a few hours. Complex solid phase diagrams are easily studied. The success or failure of fractionation techniques can be predicted from single thermograms if several phase transitions are present.     

Some applications of differential scanning calorimetry for the analysis of polymers

Applications of differential scanning calorimetry (DSC) have continued to grow in many diverse areas. It is the purpose of this paper to discuss some of the newer applications of DSC for the analysis of polymers. Applications include: heat-sealing characteristics of polyethylene, determination of additives, the cross-linking of polyethylene by peroxides and the analysis of impact polystyrene.     

Critical thermodynamic analysis of differential scanning calorimetry for studying chemical kinetics

A critical thermodynamic analysis of differential thermal calorimetry is reported herein to gain further insight into the phenomena leading to the reported differences between kinetic parameters extracted from isothermal DSC methods and those from dynamic DSC methods. The sources have been identified for the variations observed in the total heat of reaction as a function of the heating rate in dynamic DSC experiments. The analysis clearly indicates that these variations are, in fact, to be anticipated. The relationships necessary for extracting kinetic data from both isothermal and dynamic experiments are derived rigorously by resorting to classical thermodynamics. This work was supported by the Cooperative State Research Service, U.S. Department of Agriculture, under Agreement No. 92-37500-7911.     

Compatibility study of tobramycin and pharmaceutical excipients using differential scanning calorimetry,FTIR, DRX,and HPLC

Differential scanning calorimetry (DSC), isothermal stress testing–Fourier transform infrared spectroscopy (IST–FTIR), isothermal stress testing–high-performance liquid chromatography, and powder X-ray diffraction (PDRX) were used as screening techniques for assessing the compatibility of tobramycin with some currently employed ophthalmic excipients. In the first phase of the study, DSC was used as a tool to detect any interaction. The absolute value of the difference between the enthalpy of the pure tobramycin melting peak and that of its melting peak in the different analyzed mixtures was chosen as a parameter of the drug–excipient interaction degree. DSC results demonstrated that benzalkonium chloride, monobasic sodium phosphate, boric acid, edetate disodium, sodium metabisulfite, thimerosal, and potassium sorbate interact with tobramycin. Taking into account these results, it could be suggested that some of the changes observed in the IST–FTIR spectra of binary blends of tobramycin and some of the excipients would account for a possible interaction between the mixture component. In this study, PDRX did not provide much information, since only tobramycin–thimerosal interactions could be detected. DSC and IST–FTIR are suitable and simple methods for the detection of potential incompatibilities between active pharmaceutical ingredient (API) and excipients.

     

Simultaneous differential scanning calorimetry and small-angle x-ray scattering

A method is described for performing simultaneous measurements of small-angle x-ray scattering and differential scanning calorimetry. The experiment is made possible through a combination of the high flux afforded by the storage ring at the Stanford Synchrotron Radiation Laboratory, a linear position-sensitive detector with rapid response time, and a differential scanning calorimeter developed for optical microscopy. The feasibility of the technique is illustrated by examining the melting and crystallization of a polyethylene specimen. This example demonstrates the power of the technique and the accuracy and reliability of the scattering and thermal data.     

Design and development in self-reference differential scanning calorimetry

An intermediate range (50–1000°C) self-referencing differential scanning calorimeter (SR-DSC) has been built and its performance evaluated. The SR-DSC measures heat flow across a heat flow metal plate, and any changes to the heat flow caused by a thermal transition occurring in a centrally placed sample is monitored by a temperature difference across the plate. The criteria for high sensitivity are that the circular plate should be as thin as possible and have a low thermal conductivity. The best sensitivity conducive with robust behaviour was achieved with an inconel thermal plate of uniform thickness, 75 m, this gave reproducible results, and the enthalpy of the thermal transition was proportional to sample mass. Calorimeter sensitivity decreased with increasing temperature and a sloped baseline was observed. Both of these effects can be corrected mathematically. An example of the use of the SR-DSC in polymer characterisation was limited to a study of the physical ageing of PET.This revised version was published online in November 2005 with corrections to the Cover Date.Paper was read at the TAC2001 Conference in Liverpool.     

Adsorption-induced conformational changes of antifreeze glycoproteins at the ice/water interface

The conformation of antifreeze glycoprotein (AFGP) molecules adsorbed at the ice/water interface was studied by attenuated total reflection (ATR)-FTIR spectroscopy. Measurements were carried out for AFGP/D2O solution films formed on the surface of an ATR prism as a function of temperature. Using the FTIR spectrum from the O-D stretching band of D2O molecules, we monitored the supercooled and frozen states of the film and measured the thickness of the quasi-liquid layer (QLL) at the ice/prism interfaces. The AFGP structure was determined for the liquid, supercooled, and frozen states of the solution film using the amide I band spectra. No noticeable differences in conformation were observed in the solution conformation from room temperature down to the 15 K supercooling studied, whereas the alpha-helical content of AFGP suddenly increased when the supercooled solution film froze at -15 degrees C. This change in conformation can increase the overall interaction between the AFGP molecules and ice surface and allow a stronger adsorption. In contrast, the alpha-helical content of AFGP in the frozen film gradually decreased with increasing temperature and finally returned to its solution-state level at the melting point of D2O ice. This gradual decrease in the alpha-helix content directly correlates with the measured increase in QLL thickness. Finally, we conclude that the differences in the alpha-helix signals between the frozen and supercooled states indicate the conformational change of AFGP molecules upon adsorption at the ice/water interface, emphasizing the importance of the structure-function relationship, even for this highly flexible antifreeze.     

Kinetic analysis of isothermal decomposition of 2,4-dinitrophenylhydrazine using differential scanning calorimetry

Differential scanning calorimetry was used to study the thermal decomposition of 2,4-dinitrophenylhydrazine (DNPH) in isothermal regime. The DSC curves were carried out at several constant temperatures lower than the melting temperature. The standard isoconversional analysis of the obtained curves suggests an autocatalytic decomposition mechanism. This mechanism is also supported by the temperature dependence of the observed induction periods.     

The application of low temperature differential scanning calorimetry to polymer blend analysis

The temperature regions in which glass transitions (Tg's) occur for a series of blends of polybutadienes with natural rubber and two styrenebutadiene copolymers of different styrene contents have been studied by differential scanning calorimetry. The resulting DSC curves may be used to categorize the polybutadiene, determine the bound styrene content of the styrenebutadiene copolymer and quantify the polybutadiene content of the blends.

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Zusammenfassung Die Temperaturbereiche, in denen bei einer Reihe von Gemischen von Butadien mit natürlichem Kautschuk und bei zwei Styrol-Butadien-Kopolymeren mit verschiedenem Styrolgehalt Glastransformationen erfolgen, wurden mittels DSC untersucht. Die erhaltenen DSC-Kurven können zur Kategorisierung des Polybutadiens und zur Bestimmung des Gehalts an gebundenem Styrol in Styrol-Polybutadien-Kopolymeren und des Polybutadiengehalts der Gemische verwendet werden.

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Thanks are due to the Board of MRPRA for permission to publish this paper.     

Hydration of thermally denatured lysozyme studied by sorption calorimetry and differential scanning calorimetry

We have studied hydration (and dehydration) of thermally denatured hen egg lysozyme using sorption calorimetry. Two different procedures of thermal denaturation of lysozyme were used. In the first procedure the protein was denatured in an aqueous solution at 90 degrees C, in the other procedure a sample that contained 20% of water was denatured at 150 degrees C. The protein denatured at 90 degrees C showed very similar sorption behavior to that of the native protein. The lysozyme samples denatured at 150 degrees C were studied at several temperatures in the range of 25-60 degrees C. In the beginning of sorption, the sorption isotherms of native and denatured lysozyme are almost identical. At higher water contents, however, the denatured lysozyme can absorb a greater amount of water than the native protein due to the larger number of available sorption sites. Desorption experiments did not reveal a pronounced hysteresis in the sorption isotherm of denatured lysozyme (such hysteresis is typical for native lysozyme). Despite the unfolded structure, the denatured lysozyme binds less water than does the native lysozyme in the desorption experiments at water contents up to 34 wt %. Glass transitions in the denatured lysozyme were observed using both differential scanning calorimetry and sorption calorimetry. Partial molar enthalpy of mixing of water in the glassy state is strongly exothermic, which gives rise to a positive temperature dependence of the water activity. The changes of the free energy of the protein induced by the hydration stabilize the denatured form of lysozyme with respect to the native form.     

Thermal polymerization of acrylamide by differential scanning calorimetry

Thermal polymerization of acrylamide was studied by differential scanning calorimetry. Latent heat of fusion ΔH_f and enthalpy of polymerization ΔH_p values were found to be 36 and ?18.0 kcal mol^?1, respectively. The overall activation energy E for the polymerization was calculated to be 19 k cal mol^?1 up to 60% conversion. The added free-radical inhibitor (benzoquinone) was found to desensitize the thermal polymerization of acrylamide suggesting the polymerization to be a free-radical type. The existing rate equation for the heterogeneous bulk polymerization in the presence of initiators has been modified for the thermally initiated bulk polymerization of acrylamide. The experimental overall E value was found to agree well with the calculated E value when considering only the propagation and termination steps, thereby suggesting the process to be similar to postpolymerization of acrylamide.     

High-sensitivity modulation differential scanning calorimetry of protein denaturation

Method of high-sensitivity modulation differential scanning calorimetry was applied for investigation of the kinetically controlled irreversible thermal denaturation of the trypsin inhibitor from soybeans (Kunitz inhibitor, KI) in diluted solution. The measurements were carried out with a temperature-modulation capillary nanocalorimeter designed and produced by the Institute of Biological Instrumentation of the RAS (Pushchino, Russia). An algorithm of the experimental data processing and corresponding software were developed. It was shown that the modulation nanocalorimetry allows one to obtain in one experiment the temperature dependence of the rate constant for irreversible protein denaturation. The temperature dependence of the rate constant and the activation energy of the irreversible denaturation of Kunitz inhibitor were determined. The obtained value of the activation energy (E _a = 206 ± 6 kJ mol^?1) agrees with independent estimates of this kinetic parameter.     

The limit of detection in differential scanning calorimetry

A procedure is described to determine the limit of detection of DSC instruments by using tiny signals from spontaneous polymorphic transitions of CsCl, K₂Cr₂O₇ and Na₂SO₄. It is shown how such signals can be found well-resolved in DSC diagrams of powder samples. To distinguish them from the baseline noise they should exhibit a height at least twice that of the baseline width. For the instrument employed the corresponding smallest amount of heat, i.e., the limit of detection, was found to be 0.1 mJ.The authors thank Mr. H. Maltry for technical help and the Deutsche Forschungsgemeinschaft for support.