How to Determine the void volume in HPLC

How To Determine The Void Volume In HPLC

As a rule of thumb, the void volume of a system can be determined by injecting a standard without or with very little retention on a particular phase (Uracil in RP-HPLC). A slight amount of variation in this value may be explained by your unique system configuration and setup.

To determine the actual void volume of the system and column, multiply the elution time of the unretained compound by the flow rate. By subtracting the volume of the column void, you could determine the column void volume alone.

This is all about how to determine the void volume in HPLC in pharmaceutical quality control.

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