Serum aldo by LC-MS 2025 Heanue for rebuttal.prism

<b>LC-SM for serum aldosterone </b>Serum aldosterone was determined by LC-MS in mice administered vehicle and aldosterone. Steroids were extracted using a single-phase butanol:methanol extraction of 100uL of serum. 1ml of 1:1 butanol:methanol was added to each sample to precipitate proteins. The organic phase containing the steroids was dried down and reconstituted in 50% methanol in water and sonicated in a batch sonicator for 15 minutes prior to injection on an Agilent 6495C triple quad mass spectrometer coupled with an Agilent 1290 HPLC.