LC/MS/MS Analysis of Steroid Hormones in Plasma on Ascentis® Express C18 after Sample Prep using HybridSPE®-Phospholipid

LC/MS/MS Analysis of Steroid Hormones in Plasma on Ascentis® Express C18 after Sample Prep using HybridSPE®-Phospholipid

Conditions

SPE tube/cartridge HybridSPE®-Phospholipid, 96-well plate, 50 mg/well (575656-U)
sample addition To each well add 100 µL of plasma followed by 300 µL of precipitation solvent (1% formic acid or 0.5% citric acid in acetonitrile).  (Agitate via vortex for 4 minute, place on vacuum manifold and apply 10" Hg vacuum for 4 minutes. Collect filtrate and analyze directly.)
column Ascentis® Express C18, 10 cm x 2.1 mm I.D., 2.7 µm particles (53823-U)
column temp. 50 °C
mobile phase [A] 5 mM ammonium formate pH 4.0 with formic acid; [B] methanol
gradient 60% B for 3 min, 60% B to 95% B in 5 min, held at 95% B for 2 min
flow rate 0.3 mL/min
injection 2 µL
detector ESI+, MRM

Description

Analysis Note This application shows the separation of steroid hormones in plasma using LC/MS/MS. HybridSPE-Phospholipid removed endogenous phospholipids and precipitated proteins, thereby allowing rapid, sensitive analysis by LC/MS/MS using an Ascentis Express C18 column. The highest grade LC-MS solvents were used to supply low background interference and low particulate contaminants for robust, trouble-free operation. Cerilliant CRMs provided reliable identification and quantification.
Categories Analytical Chromatography, Pharmaceutical Analysis, Clinical, SPE Applications, antiepileptic/antiseizure, hormones, Anticonvulsant/antiepiletic/antiseizure, Hormones
Featured Industry Clinical
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC, HybridSPE is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for HPLC

Materials

     
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