Determination of Olopatadine in Human Tears by Hydrophilic Interaction Liquid Chromatography–MS/MS Method

Abstract

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

Financial & competing interests disclosure

The authors thank the Ministry of Education, Science and Technological Development of the Republic of Serbia for supporting these investigations in Project Number 172052. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The clinical study was approved by the Ethics Committee of the Clinical Center of Serbia on 24 April 2016. Signed informed consent forms were received from all participants after the aims and risks of the study were explained. The tear samples were obtained from 50 outpatients with allergic conjunctivitis aged from 18 to 60 following topical ocular administration of 0.1% olopatadine hydrochloride in both eyes. Each human tear sample was collected using sterile Schirmer strips painlessly and with minimal trauma to the patient. Collecting time was limited to 5 min. At the time and 15 min after instilling, subjects did not wear contact lenses. A single tear sample was collected immediately after eye drops instillation and at 0.25, 0.5, 1, 2 and 8 h postdose.