LC/MS/MS Analysis of Estradiol (Underivatized) from Patient Serum on Titan™ C18, 5 cm Column

LC/MS/MS Analysis of Estradiol (Underivatized) from Patient Serum on Titan™ C18, 5 cm Column


sample preparation SPE (Solid Phase Extraction)
column Titan C18, 5 cm x 2.1 mm I.D., 1.9 µm particles (577122-U)
column temp. 40 °C
mobile phase [A] 0.2 mM ammonium fluoride; [B] methanol
gradient 40 to 82%B in 4.6 min; to 100%B in 0.05 min; held for 1.15 min; to 40% B in 0.05 min; held for 1.65 min
flow rate 0.4 mL/min
sample A 200 µL sample of calibrator or serum was mixed with 50 µL of internal standard solution (1 nmol/L 13C4-estradiol in 30% methanol) and vortexed. (Liquid-liquid extraction was performed by addition of 1 mL of MTBE. Supernatants were dried down and reconstituted in 100 µL of 50% methanol before being transferred to the LC/MS/MS system.)
sample patient serum sample containing estradiol, 56 pmol/L
injection 20 µL
detector MS, ESI(-), MRM transitions listed on figure


Analysis Note A fast and robust LC/MS/MS method was developed for the measurement of estradiol in human serum using Titan C18 1.9 μm columns. Ultra high purity LC-MS solvents were used to supply low background interference and low particulate contaminants for robust, trouble-free operation. Cerilliant certified reference material grade standards provided reliable quantification.
Categories Analytical Chromatography, Clinical, Hormones, hormones
Featured Industry Clinical
Legal Information Titan is a trademark of Sigma-Aldrich Co. LLC
suitability application for LC-MS


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