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Besides having numerous cellular functions, GSH is an important antioxidant and is considered a valuable biomarker in evaluating oxidative stress. This paper provides a sensitive analytical method using HPLC\u2010ECD to quantify GSH in erythrocytes, validated using the ICH guidelines for Bioanalytical Method Validation. The sample preparation was optimised using centrifugal filtration and a hypotonic phosphate buffer for extracting GSH from erythrocytes. HPLC\u2010ECD parameters were adjusted to allow a fast, reversed phase, isocratic separation in 10\u00a0min. The detector response was linear between 0.3 and 9.5\u00a0\u03bcg\/mL with a satisfactory regression coefficient and a LOQ of 0.11\u2009\u03bcg\/mL. Intra\u2010 and inter\u2010day repeatability ranged between 1.10% and 8.57% with recoveries ranging from 94.3% to 106.0%. Dilution integrity, benchtop, freeze\u2013thaw and long\u2010term stability were investigated. Samples were stable for up to 6\u00a0months at \u221280\u00b0C. This method has a good linear response and is repeatable, precise and accurate. It minimises GSH auto\u2010oxidation using a centrifugal filter during sample preparation, instead of acidification. Therefore, this analytical method is suitable for quantifying GSH in erythrocytes as a marker of oxidative stress.<\/jats:p>","DOI":"10.1002\/bmc.6021","type":"journal-article","created":{"date-parts":[[2024,10,1]],"date-time":"2024-10-01T21:29:48Z","timestamp":1727818188000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Validation and optimisation of reduced glutathione quantification in erythrocytes by means of a coulometric high\u2010performance liquid chromatography analytical method"],"prefix":"10.1002","volume":"38","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6525-5176","authenticated-orcid":false,"given":"Stef","family":"Lauwers","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences, Natural Products &amp; Food Research and Analysis \u2013 Pharmaceutical Technology (NatuRAPT) University of Antwerp  Antwerp 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