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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