This unit describes dynamic methods for monitoring the gelation process of protein dispersions and for determining rheological properties of the final gels. In a dynamic test, a test material is subjected to a controlled oscillation, the frequency of which determines the rate of deformation of the sample. The magnitude of deformation is kept very small to prevent destruction of gel structures during measurement. Two basic parameters reflecting elastic (G′; storage modulus) and viscous (G″; loss modulus) components of rheological properties are obtained at various frequencies. This frequency range will depend on the sample type and the purpose of the study (see Critical Parameters and Troubleshooting). The measuring instruments are operated by computers and the experimental procedure is simple. UNIT H3.1 discusses much of the theory behind dynamic rheometer tests.
MaterialsSample protein solution or gel Chemical substance to induce gelation, if needed Immiscible reagent for preventing solvent evaporation (e.g., mineral or silicone oil), if needed Dynamic rotational rheometer with appropriate test fixture Computer with software package to control rheometer 1. Attach an appropriate test fixture to a dynamic rotational rheometer and connect rheometer inline with a computer.The appropriate text fixture type (or geometry) and size depend on the substance that is being analyzed. UNIT H1.2 includes a discussion on choosing a geometry. The amount of sample needed will depend on the rheometer and test fixture that are used. Ideally, there should not be excess sample (e.g., below or above the inner cylinder or outside the upper plate). Usually, sample below or above the inner cylinder does not contribute significantly to the results because of its smaller contact area compared with that of the wall of the cylinder. For steady rotational viscometry, it is often critical to cut the sample outside the upper plate. Thus it is also recommended to do so in dynamic tests. Changes in sample volume that often occur during gelation must also be considered.2a. For a gel induced by a chemical substance: Mix a sample protein solution with a chemical substance to induce gelation, place the mixture in the test fixture, and cover it with an immiscible reagent.The chemical substance required to induce gelation, as well as the concentrations of chemical and protein, will need to be determined empirically.2b. For a gel induced by heating: Place a sample protein solution in the fixture, cover it with an immiscible reagent, and use the computer to apply the appropriate temperature to the fixture for an appropriate length of time.The time and temperature of the incubation, as well as the protein concentration, will need to be determined empirically. Depednding on the purpose of the study, a temperature ramp or constant temperature can be used here and in steps 3 to 5.2c. For a preformed gel: Place a sample gel in the fixture and cover it with an immiscible reagent, if necessary. Continue with step 4.3. Set the computer to display G′ and G″. Moni...