Acyclovir and hydrocortisone often appear in the state of a mixture. In the principle, mixture significantly different polarity are easy to resolve on reverse phase HPLC columns. In this situation, gradient elution is usually required to obtain a reasonable retention time for all mixture. However the gradient is not always desirable especially for high through put mode of operation. so acyclovir and hydrocortisone can be separation by HPLC.

Note:

1. Additional work is required to prepare two different solvents for the mobile phase.
2. Long operating time .
3. Before each injection sample, the chromatography column equilibration takes a long time.

A mixture of acyclovir and hydrocortisone is a sample. Those are alway constituted in topical creams. Acyclovir is a polar molecule (logP = – 1.2), while hydrocortisone (logP = ~1.7) is a hydrophobic compound. Their separation on a reverse phase 150 mm, 5ìm column would take 10+ minutes. The same separation, but much faster, can be obtained on a Mixed-Mode (Primesep 100) column with simple isocratic mode of elution. For LC-MS compatible conditions the sulfuric acid can be substituted by TFA or ammonium formate.

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