Some Practical Considerations when Determining the Void Volume in High-Performance Liquid Chromatography

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Abstract
The void volume of a reversed-phase high-performance liquid chromatography column is determined by extrapolation of the retention time of a homologous series of alkylbenzenes as well as by the use of a number of void markers. It is found that the use of void markers such as uracil or D2O provides a simple, rapid, and reliable procedure for the analyst who needs a reasonably good estimate of the value. Extrapolation of the retention times of a series of homologous series of alkylbenzenes leads to a larger value of the void volume than does the use of a void marker. In addition, the extrapolation technique is found to be inappropriate for eluents containing THF, and the measured value is dependent on the mathematical approach used to obtain the void volume.

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