Protocol for Aqueous Humor Metabolite Measurements for Glaucoma Research
- Authors: Anna Mueller1, Isabella Moceri2, Sanjoy K. Bhattacharya3
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View Affiliations Hide Affiliations1 Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL 33136, USA 2 Miami Integrative Metabolomics Research Center, Miami, FL, 33136, USA 3 University of Miami Miller School of Medicine, Miami, 1638 NW 10th Avenue, Suite 707A, FL 33136, USA
- Source: Research Protocols for Ophthalmic Disease Mechanisms and Therapeutics: Glaucoma - Ocular Hypertension , pp 541-549
- Publication Date: August 2025
- Language: English
Protocol for Aqueous Humor Metabolite Measurements for Glaucoma Research, Page 1 of 1
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Ultra-high-performance liquid chromatography-mass spectrometry is commonly used in the untargeted analysis of metabolites within mammalian aqueous humor samples. Aqueous humor is a transparent fluid that bathes the anterior segment of the eye to nourish the cornea and lens. Aqueous humor also assists in the maintenance of intraocular pressure. In glaucoma, aqueous humor flow is impeded, leading to ocular hypertension and optic nerve damage. The application of metabolomics to study aqueous humor allows for advancements in knowledge of the molecular mechanisms in glaucoma. In this chapter, we put forth a protocol using hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass spectrometry to accurately identify metabolomic data of the aqueous humor samples. The purpose of this chapter is to provide an extraction and identification method of mammalian aqueous humor metabolites using liquid chromatography-tandem mass spectrometry.
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