A validated sensitive LC-MS/MS method and its application in elucidating the unique ocular pharmacokinetic profile of 0.01% atropine underpinning its clinical utility for myopia.

Juan Yue; Susu Liu; Shuaibing Zhou; Yanglei Yuan; Junlu Dong; Shoujun Jian; Yonghui Wei; Siyuan Qi; Yanting Xie; Hongmin Zhang

doi:10.1016/j.jchromb.2026.125245

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A validated sensitive LC-MS/MS method and its application in elucidating the unique ocular pharmacokinetic profile of 0.01% atropine underpinning its clinical utility for myopia.

Juan Yue·Susu Liu·Shuaibing Zhou·Yanglei Yuan·Junlu Dong·Shoujun Jian·Yonghui Wei·Siyuan Qi·Yanting Xie·Hongmin Zhang

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Abstract

A sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated to quantify atropine in ten rabbit ocular tissues enabling systematic characterization of the ocular pharmacokinetic profile of 0.01% atropine sulfate eye drops after a single topical administration. The method demonstrated excellent linearity (coefficient of determination, R2 ≥ 0.9908) across all matrices, with lower limits of quantification (LLOQ) of 0.05 ng/mL for most tissues and 0.10 ng/mL for retina and lens; intra- and inter-day accuracy, precision, matrix effects, extraction recoveries, and stability all met the acceptance criteria. Following a single bilateral topical dose (50 μL/eye) in New Zealand White rabbits, atropine distributed rapidly into all 12 ocular compartments (the sclera further divided into three anatomical regions) with marked heterogeneity-the highest exposures were found in conjunctiva and cornea, a distinct anterior-to-posterior concentration gradient was observed in the sclera, sustained retention was noted in the retina (mean residence time from zero to the last measurable time point, MRT0-t 3.30 h), while aqueous and vitreous humor eliminated rapidly (elimination half-life, t₁/₂ < 0.7 h), and all tissues except aqueous humor followed a two-compartment model. This validated method and the comprehensive pharmacokinetic data reveal that topically applied 0.01% atropine achieves sustained exposure in key myopia-regulating tissues (retina, choroid, posterior sclera) with low exposure in side-effect target tissues (iris, ciliary body, lens).

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Animals Atropine Rabbits Tandem Mass Spectrometry Liquid Chromatography-Mass Spectrometry Reproducibility of Results Ophthalmic Solutions Myopia Eye Linear Models Tissue Distribution Chromatography, Liquid 0.01% atropine sulfate eye drops LC-MS/MS Myopia control Ocular pharmacokinetics Ocular tissue distribution Journal Article

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Juan Yue, Susu Liu, Shuaibing Zhou, Yanglei Yuan, Junlu Dong, Shoujun Jian, Yonghui Wei, Siyuan Qi, Yanting Xie, Hongmin Zhang. 2026-08-09. A validated sensitive LC-MS/MS method and its application in elucidating the unique ocular pharmacokinetic profile of 0.01% atropine underpinning its clinical utility for myopia.. https://doi.org/10.1016/j.jchromb.2026.125245

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