High Throughput Automated Determination of Glutathione Based on the Formation of a UV-Absorbing Thioacrylate Derivative

Author(s): Constantinos K. Zacharis, Paraskevas D. Tzanavaras

Journal Name: Combinatorial Chemistry & High Throughput Screening
Accelerated Technologies for Biotechnology, Bioassays, Medicinal Chemistry and Natural Products Research

Volume 13 , Issue 6 , 2010

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Abstract:

Sequential injection analysis (SIA) has been employed as the platform for the development of a high throughput automated assay for glutathione (GSH) in nutrition supplements. The proposed assay is based on the on-line formation of a UV-absorbing thioacrylate derivative (δmax = 285 nm) upon reaction of thiol with the new reagent ethylpropiolate (EP). This is the very first application of this reagent for the determination of GSH either in batch or continuous flow modes and the first SI method for the determination of GSH in pharmaceutical supplements. The various variables that typically affect such a protocol were studied, while thorough validation experiments according to established standards proved the reliability of the assay for the intended applications. As many as 100 sample injections were enabled per hour.

Keywords: High throughput assay, glutathione, sequential injection, ethyl propiolate, UV-spectrophotometry

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Article Details

VOLUME: 13
ISSUE: 6
Year: 2010
Page: [461 - 468]
Pages: 8
DOI: 10.2174/138620710791515923
Price: $65

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