1. Choose an isocratic mobile phase.  The type of column being used and nature of the analyte should be considered.  Polymeric columns generally require a higher concentration of organic modifier than silica or zirconia based columns. The polarity, functionality, structure, pKa and reactivity of the analyte must also be considered.  The amount of organic modifier in the mobile phase can generally be reduced by about 10-20% at 50°C when compared to room temperature. 
2. Choose the proper mobile phase buffer. In many cases, water without a buffer will provide an adequate separation. Selection of mobile phase pH should be based on the pKa of the analyte of interest. There are several good sources for information on obtaining pKa information and selecting mobile phase buffers.
3. Run a temperature gradient from 35°C to the maximum recommended temperature for the column you are using.  Hold at the maximum temperature for five minutes.  Use a ramp rate of 10°C or 15°C per minute. 
4. Based upon the results, decide if you want an isothermal or a temperature gradient method. 
• For an isothermal method, optimize temperature, and adjust the mobile phase composition to achieve the best separation.  Optimize the flow rate to achieve the shortest analysis with adequate resolution and efficiency. 
• For a temperature gradient method, choose a starting temperature and adjust the mobile phase composition, then optimize ramp rate and flow rate.

To replace an existing binary solvent gradient with a programmed temperature gradient.

1. Choose an isocratic mobile phase composition approximately midway between the starting and ending compositions of the solvent gradient.  For example, if you are running a linear gradient from 20% to 100% acetonitrile, start with an isocratic mobile phase of 40% water and 60% acetonitrile. 
2. Run a temperature gradient from 35°C to the maximum allowable temperature for the column you are using.  Hold at the maximum temperature for at least five minutes to ensure elution of all components. Use a ramp rate of 10°C or 15°C per minute.
3. Based on the results, choose a starting temperature then adjust the mobile phase composition as necessary.  Optimize ramp rate and flow rate to achieve the shortest analysis time with adequate resolution and efficiency.

To speed up an existing isocratic method or reduce the amount of organic modifier in the mobile phase:

1. Start with the existing isocratic method.  Perform the analysis at 50°C. 
2. Increase the temperature in 50°C increments, reducing the amount of organic modifier in the mobile phase as necessary to produce a good separation. 
3. Optimize the temperature, mobile phase composition and flow rate to achieve the shortest analysis time with adequate resolution and efficiency.