Biochemical Techniques and Instrumentation

2.6.3.5. Thermostatic oven

The column is contained in a thermostat-controlled oven. The separation of compounds in the column through the partitioning behavior is dependent on the oven temperature. The optimum column temperature is fixed based on the boiling point of the compounds in the sample. The separation of compounds is achieved by two types of programming:

1. Isothermal programming
2. Gradient programming

In isothermal programming, the oven is set at same temperature and different components are eluted. In gradient programming, the oven temperature is gradually increased over time to elute the high–boiling point components with a wide range of boiling points. A temperature slightly above the average boiling point of the sample results in an elution time within 30 min. The use of minimal temperatures give good resolution, but increase elution time. The temperature programming is useful, if the components in the sample have wide boiling points.