Aim: Dexamethasone (Dex) has been used for the treatment of ocular diseases, presenting concentrations up to 150 ng/ml in the biological matrix. Drug sampling has been performed from aqueous humor (AH), which the volume varies from 50 to 200 μl, becoming a challenge for analytical analyses.
Results: We developed and validated a direct and sensitive method by LC-MS/MS for Dex measurement in human AH, using water as surrogate matrix to reduce the sample volume applied in some validation assays. With accuracies of 99.6% and precision within 15% in the range of 0.1-150 ng/ml, Dex had an 87-day stability.
Conclusion: Our method is robust and sensitive enough to be applied in bioequivalence studies with human AH using only 20 μl of biological sample.
Keywords: LC–MS/MS; aqueous humor; dexamethasone; direct method; small volume; validation.