【摘要】 利用新型荧光试剂1,2-苯并-3,4-二氢咔唑-9-乙基肼基甲酸酯(BCEC)作柱前衍生化试剂,在HypersilBDSC18(4.6mm×200mm,5μm)色谱柱上,采用梯度洗脱对皮质醇、皮质酮、睾酮、孕酮4种类固醇荧光衍生物进行了优化分离。65℃下在乙腈溶剂中以三氯乙酸作催化剂,衍生反应2h后获得稳定的荧光产物。激发和发射波长分别为333nm和390nm。采用大气压化学电离源(APCI)正离子模式,实现了黑线姬鼠粪便中4种类固醇化合物的定性及定量测定。线性回归系数均在0.9999以上,检出限为47.3~71.2fmol。更多还原

【Abstract】 A simple and sensitive method for the determination of steroid compounds extracted from rat feces using 1,2-benzo-3,4-dihydrocarbazole-9-ethyl-carbonylhydazine (BCEC) as pre-column derivatization reagent by high performance liquid chromatography in combination with a gradient elution with fluorescence detection and mass spectrometric identification has been developed. Studies on derivatization conditions indicate that steroid compounds react with BCEC at 65℃ within a two-hour period in the presence of trichloroacetic acid (TCA) catalyst with acetonitrile as reaction co-solvent to give the corresponding sensitive fluorescence derivatives. The fluorescence excitation and emission wavelengths are 333 nm and 390 nm, respectively. The identification of steroid derivatives is performed under atmospheric-pressure chemical ionization (APCI) source in positive-ion mode. Correlation coefficients for steroid derivatives are >0.9999, and detection limits (at signal-to-noise ratio of 3∶1) are between 47.3 to 71.2 fmol. The established method is sensitive and reproducible for the determination of steroid compounds from real samples with satisfactory results.更多还原