Determination of estradiol and estrone concentrations in human plasma by UPLC-MS/MS method
Objective To establish a method for simultaneous determination of estradiol(E2)and estrone(E1)concentrations in plasma by ultra-high performance liquid chromatography-mass spectrometry(UPLC-MS/MS).Methods The sample was pretreated by liquid-liquid extraction,and the internal standard was 17β-estradiol-D3.The chromatographic column was ACQUITY UPLC BEH C18(2.1 mm x 100.0 mm,1.7 μm),and the mobile phases were:100％water containing 0.2 mmol of ammonium fluoride-100％methanol containing 0.2 mmol of ammonium fluoride,with a flow rate of 0.30 mL·min-1,column temperature:30 ℃,autosampler temperature:15 ℃,injection volume:6 µL.The method was monitored by an electrospray ionisation source in negative ion mode with multiple reactions.The method was investigated for its specificity,standard curve and lower limit of quantification,precision and recovery,matrix effect and stability.Results Both estradiol and estrone were within 20-1 000 pg·mL-1 with good linearity and the standard curves were y=1.87×10-3x+3.71 × 10-2(R=0.999 27)for estradiol and y=8.48×10-3 x+6.46×10-2(R=0.998 90)for estrone,the lower limit of quantification was 20 pg·mL-1 for both estradiol and estrone.The intra-day and inter-day relative standard deviations of estradiol and estrone were less than 9.46％.The recoveries ranged from 97.64％to 107.35％,and the matrix effects ranged from 93.29％to 109.87％.Conclusion The method is proprietary,sensitive and suitable for therapeutic drug monitoring studies of estradiol and estrone.
estradiolestroneultra-high performance liquid chromatography-tandem mass spectrometerpharma-cokinetics
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