HPLC Quantification of Redox Markers

Tissue and plasma Cys, CySS, GSH, and GSSG were assayed by HPLC as S-carboxymethyl, N-dansyl derivatives using γ-glutamylglutamate as an internal standard as previously described [17 (link),18 (link)]. The HPLC system consisted of an aminopropyl column, 2 Waters 515 HPLC Pumps to deliver a gradient of acetate and methanol, a Waters 717 plus Autosampler with refrigeration unit, and a Waters 2475 Multi λ Fluorescence Detector. Integration of peak areas and comparison to the internal standard was performed by Waters Empower 3 software. Plasma: (WT vehicle: n = 5), (NHERF1 KO vehicle: n = 5), (WT cisplatin: n = 7), and (NHERF1 KO cisplatin: n = 9). Kidney: (WT vehicle: n = 5), (NHERF1 KO vehicle: n = 5), (WT cisplatin: n = 8), and (NHERF1 KO cisplatin: n = 9).