Session: MON 723-757-Renin-Angiotensin-Aldosterone System/Endocrine Hypertension
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Poster Board MON-737
The analysis of aldosterone in serum was accomplished using UHPLC coupled to the AB SCIEX Triple Quad™ 6500 system. The method employs the Multiple Reaction Monitoring (MRM) scan mode for the detection of aldosterone, using negative electrospray ionization (ESI). Pre-analytical sample preparation consisted of liquid-liquid extraction in MTBE, followed by evaporation under N2 gas, and reconstitution of the sample in the LC mobile phase. Chromatographic separation was accomplished using a Phenomenex Gemini-NX C18 analytical column (150x3.0mm, 5um). A gradient elution was employed, consisting of water + 2mM ammonium acetate (mobile phase A) and methanol + 2mM ammonium acetate (mobile phase B), at a flow rate of 500uL/minute. The total run-time for the method including re-equilibration was 10 minutes, to ensure adequate separation of the aldosterone analyte from endogenous interferences.
The method described here was used to analyze a series of human serum samples containing concentrations of aldosterone ranging from 14 pg/mL to 300 pg/mL. The limit of quantitation for aldosterone in serum was observed to be less than 1 pg/mL. In developing the method, great care was taken to ensure that the analyte of interest does not elute in a time window where potential interferences also elute. The method displayed excellent linearity over the concentration range from 1-1000 pg/mL (r=0.99971). Accuracies ranged from 89-118% over the entire concentration range from 1-1000 pg/mL of aldosterone, and the CV% ranges from 0.5-9.1%. The accuracy and CV% for the lowest calibrator, at 1 pg/mL, were 100% and 8.7%, respectively.
Disclosure: MJYJ: Employee, AB SCIEX. MW: Employee, AB Sciex.
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