HPLC-based quantitative assessment of antiviral agents in artificial saliva.

BACKGROUND

The efficient analytical method for antiviral drug quantification is crucial in pharmaceutical and biomedical research, particularly for ensuring accurate dosage and monitoring therapeutic efficacy. Accurate quantification of these compounds in artificial saliva is essential for pharmacokinetic studies, as saliva-based drug monitoring offers a noninvasive alternative to traditional blood sampling.

OBJECTIVE

A novel high-performance liquid chromatographic (HPLC) method for simultaneous analysis of three selected antivirals: Tenofovir Disoproxil Fumarate (TDF), Favipiravir (FAV), Ritonavir (RIT) and internal standard as another antiviral (emtricitabine) in artificial saliva was optimized and validated, including sample preparation using new extraction procedure.

METHODS

The chromatographic separation was achieved with XTerra, C18 (250 × 4.6 mm, 3.5 µm) analytical column (20 °C) at a wavelength of 215 nm, and the mobile phase was composed of 0.1% phosphoric acid, methanol, and an acetonitrile mixture; 32:60:8 (v/v) at a flow rate of 0.8 mL/min. The method was linear over the range of 0.1-1.0 µg/mL.

RESULTS

The method has been found according to ICH guidelines specific, accurate (recovery, 93.003%-103.357%), sensitive (LOD and LOQ were values found as 0.03 and 0.1 µg/mL, respectively), and precise (RSD; 0.412%-3.175%).

CONCLUSION

A validated HPLC method was developed for the precise determination of different antiviral agents in artificial saliva.