Products Cited in Publications Worldwide Development of a HPLC-MS/MS method for assessment of thiol redox status in human tear fluids

Development of a HPLC-MS/MS method for assessment of thiol redox status in human tear fluids

Wu, Jiandong; Sigler, Austin; Pfaff, Annalise; Cen, Nan; Ercal, Nuran; Shi, Honglan

DOI: PMID:

Abstract

Oxidative stress is reported to be part of the pathol. of many ocular diseases. For the diagnosis of ocular diseases, tear fluid has unique advantages. Although numerous anal. methods exist for the measurement of different types of biomols. in tear fluid, few have been reported for comprehensive understanding of oxidative stress-related thiol redox signaling. In this study, a high-performance liquid chromatog.-tandem mass spectrometry (HPLC-MS/MS) method was developed to determine a panel of twelve metabolites that systematically covered several thiol metabolic pathways. With optimization of MS/MS parameters and HPLC mobile phases, this method was sensitive (LOQ as low as 0.01 ng/mL), accurate (80-125% spike recovery) and precise (<10% RSD). This LC-MS/MS method combined with a simple tear fluid collection with Schirmer test strip followed by ultrafiltration allowed the high-throughput anal. for efficient determination of metabolites associated with thiol redox signaling in human tear fluids. The method was then applied to a small cohort of tear fluids obtained from healthy individuals. The method presented here provides a new technique to facilitate future work aiming to determine the complex thiol redox signaling in tear fluids for accurate assessment and diagnosis of ocular diseases.

Keywords

Biomarker ; Glutathione ; HPLC-MS/MS ; Ocular disease ; Tear fluid ; Thiol

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