Aim: The objective of the study was development of hydrophilic interaction liquid chromatography–ESI/MS/MS method for the determination of olopatadine in tear matrix. Materials & methods: Separation was performed on Acquity BEH amide column (2.1 × 100 mm, 1.7 μm). The mobile phase was consisted of 0.1% formic acid in water and acetonitrile. Mianserin hydrochloride was implemented as an internal standard. The artificial tear fluid was used as matrix. The tear samples were collected using Schirmer test strips. For the optimization of ultra pressure liquid chromatography conditions, Box–Benhken design was utilized. Results: The optimal values of the ion source and collision cell parameters were found. Quantification was performed in multiple reaction monitoring mode. The optimized method was fully validated. Conclusion: The proposed method was utilized for monitoring of olopatadine in human tear.
Keywords:
- Box–Benhken design
- human tears
- hydrophilic interaction liquid chromatography
- olopatadine
- UPLC–MS/MS