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Development and application of LC-MS/MS method for the quantification of hydrogen sulfide in the eye. | LitMetric

Development and application of LC-MS/MS method for the quantification of hydrogen sulfide in the eye.

Anal Biochem

Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, Texas Southern University, Houston, TX, 77004, USA. Electronic address:

Published: April 2024

AI Article Synopsis

Article Abstract

There are limited studies that report the physiological levels of HS in the eye. The currently available UV/Vis methods lack the required sensitivity and precision. Hence, the purpose of this study was to develop and validate a sensitive and robust pre-column derivatization LC-MS/MS method to measure changes in HS levels in tissues from isolated porcine eyes. HS was derivatized and an LC-MS/MS method was developed to monitor the derivatized product, Sulfide-dibimane (Sdb) using a reverse phase Waters Acquity BEH C18 column (1.7 μm, 2.1 × 100 mm). HS quantification was performed using multiple-ion reaction monitoring (MRM) in positive mode, with the transitions of m/z 415.0 → m/z 223.0 for Sdb and m/z 353.0 → m/z 285.0 for internal standard (griseofulvin). This method provided a suitable way to quantify HS and was then successfully adapted to measure HS levels in isolated porcine iris-ciliary body tissues previously treated in the presence or absence of varying concentrations of lipopolysaccharide (LPS, 5-100 ng/ml), a pro-inflammatory agent. Isolated iris-ciliary bodies (ICB) from porcine eyes were cut into quadrants of approximately 50 mg and homogenized using a 1:3 volume of homogenizing buffer. HS in the supernatant was then derivatized with monobromobimane and quantified.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11359680PMC
http://dx.doi.org/10.1016/j.ab.2023.115448DOI Listing

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