Since the outbreak of COVID‐19, many antiviral drugs have been used to treat infected patients. Excessive drug concentrations may not only damage patients' organs, but also lead to increased mortality. The lack of data makes it difficult for them to be treated with these drugs under proper and safe dosage. However, due to the complexity of fluid matrixand structures of the antiviral drugs, it is still difficult to detect the drugs in biofluids. Herein, solid‐phase microextraction (SPME) is applied for the detection of biological matrix, which has the advantages of simplicity, efficiency and environmental protection. With abundant hydrogen bond sites and appropriate molecular volume, biocompatible graded‐pore HOF10x series materials are selected as the fiber coating for exploration. Consequently, HOF101 could achieve high enrichment effect of drugs due to the commensurate mesopores and hydrogen bond non‐covalent interaction. Moreover, the excellent extraction capacity of the HOF101 fiber is verified to be consistent in various interfering conditions. The obtained limits of detections (LODs) are 0.24–0.66 ng L−1, and the recoveries of the biofluids in three concentration levels ranged from 80.30% to 120.1%. It shows the low LODs and satisfactory recoveries of this analytical method, which is expected to be applied in medical systems for drug monitoring.