A simple and rapid HPLC method requiring small volumes (250IL) of human serum after C18 SPE sample preparation was developed using monolithic technology for simultaneous determination of all-trans-retinoic acid, 13-cis-retinoic acid, retinol, gamma- and alpha-tocopherol. Potential utilization of the method includes monitoring of drug concentrations in patients treated with retinoic acid for acute promyelocytic leukemia or for other malignancies in clinical trials.