4.5 Article

Development and validation of a direct HPLC method for the determination of salivary glutathione disulphide using a core shell column and post column derivatization with o-phthalaldehyde

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DOI: 10.1016/j.jchromb.2022.123216

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Human saliva; Glutathione disulfide; Post column derivatization; Liquid chromatography; Core shell reversed phase column

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In this study, an optimized and validated method was used to monitor glutathione disulfide (GSSG) in human saliva samples. The results showed accurate quantification of GSSG in the samples, with a concentration range of 100-2000 nmol L-1 and a LOD of 20 nmol L-1. The stability of the samples was ensured by blocking reduced glutathione with N-ethylmaleimide.
Glutathione disulfide (GSSG) has been monitored in human saliva samples by an optimized and validated method that is based on liquid chromatography coupled to on-line post column derivatization. The analyte was separated from the sample matrix using a 100% aqueous mobile phase through a core-shell reversed phase column. Following optimization of the reaction using Box-Behnken experimental design and validation, GSSG was quantified accurately and selectively in the range of 100-2000 nmol L-1 with a LOD of 20 nmol L-1. GSSG was quantified in 15 out of 20 human saliva samples (75%) with a mean value of 860 nmol L-1 (150-4600 nmol L-1). Blocking of reduced Glutathione with N-ethylmaleimide ensured stability of the samples for at least 72 h at all temperatures examined.

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