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Hypersil BDS C18 （4.6 mm×200 mm, 5 μm） 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2 h后获得稳定的荧光产物.激发和发射波长分别为333 nm和390 nm.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47.3～71.2 fmol.
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.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2006年第10期1406-1410,共5页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金资助项目(No.20075016)
关键词
高效液相色谱-质谱
荧光检测
柱前衍生
类固醇
High performance liquid chromatography/mass spectrometry, fluorescence derivatization, pre-column derivatization, steroids
