摘要
目的建立一种同时测定患者血浆中雌二醇(E2)及雌酮(E1)浓度的超高效液相色谱-质谱联用(UPLC-MS/MS)法。方法用液液萃取法对样品进行前处理,内标为17β-雌二醇-D3。色谱柱:ACQUITY UPLC BEH C18(2.1 mm×100.0 mm,1.7μm),流动相:100%水含0.2 mmol氟化铵(NH_(4)F)-100%甲醇含0.2 mmol NH4F,流速:0.30 mL·min^(-1),柱温:30℃,自动进样器温度:15℃,进样量:6μL。用电喷雾离子化源,负离子模式,多反应监测。考察该方法的专属性、标准曲线与定量下限、精密度与回收率、基质效应及稳定性。结果雌二醇和雌酮均在20~1000 pg·mL^(-1)内,线性关系良好,其标准曲线雌二醇为y=1.87×10^(-3)x+3.71×10^(-2)(R=0.99927);雌酮为y=8.48×10^(-3)x+6.46×10^(-2)(R=0.99890),定量下限均为20 pg·mL^(-1),雌二醇和雌酮批内和批间相对标准偏差均小于9.46%,回收率在97.64%~107.35%,基质效应在93.29%~109.87%。结论本方法专属性强、灵敏度高,适用于雌二醇和雌酮的治疗药物监测研究。
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×100.0 mm,1.7μm),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:6μL.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 20-1000 pg·mL^(-1)with good linearity and the standard curves were y=1.87×10^(-3)x+3.71×10^(-2)(R=0.99927)for estradiol and y=8.48×10^(-3)x+6.46×10^(-2)(R=0.99890)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.
作者
李京萌
张咏梅
LI Jing-meng;ZHANG Yong-mei(Graduate School,Hebei Medical University,Shijiazhuang 050031,Hebei Province,China;D.epartmenttof Pharmacy,Hebei General Hospital,Shijiazhuang 050051,HebeiPi rovince,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2024年第16期2392-2395,共4页
The Chinese Journal of Clinical Pharmacology
关键词
雌二醇
雌酮
超高效液相色谱-质谱联用法
药代动力学
estradiol
estrone
ultra-high performance liquid chromatography-tandem mass spectrometer
pharma-cokinetics
