同位素稀释-超高效液相色谱-串联质谱法同时测定精油中的7种雌性激素

A reliable ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method for the simultaneous determination of 7 female sex hormones(estriol,estradiol,estrone,ethinyloestradiol,dienestrol,hexestrol,diethylstilbestrol) in essential oil was developed.The sample was extracted by ethylacetate-normal hexane solution(2∶98,v/v) and the extract was purified by a silica solid phase extraction-based clean-up column.Then,the analytes were separated on an ACQUITY UPLC BEH SHELD RP18 column(100 mm×2.1 mm,1.7 μm) in gradient elution with the mobile phases of water and acetonitrile.The separated compounds were detected with a Waters Xevo TQ MS tandem quadrupole mass spectrometer operated in negative electro-spray ionization using multiple reaction monitoring mode.Estriol-D3,estradiol-D3 and diethylstilbestrol-D6 were used as the internal standards to reduce the matrix effects.The limits of detection and quantitation for the 7 female sex hormones in essential oil were 0.3-7 μg/kg and 1-20 μg/kg,respectively.Good linear relationships and high correlation coefficients(r2≥0.997) were obtained in the mass concentration range of 20-500 μg/L.The average recoveries were 88.5%-114.8% and the intra-assay relative standard deviations were 4.8%-18.9% at the spiked levels of 20-500 μg/kg.Finally,a total of 12 samples randomly collected from different supermarkets in Zhejiang Province were screened for the 7 female sex hormones by the proposed method.The results showed that only one sample contained estradiol and estrone.