高效液相色谱质谱_荧光检测_柱前衍生_类固醇_

鼠类粪便中类固醇的荧光标记及质谱鉴定

http://www.100md.com 《分析化学》 2006年第10期

高效液相色谱质谱,荧光检测,柱前衍生,类固醇,,高效液相色谱质谱,荧光检测,柱前衍生,类固醇,1引言,2实验部分,3结果与讨论,5结论,Refer

     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。

    关键词高效液相色谱质谱, 荧光检测, 柱前衍生, 类固醇

    Determination of Steroids from Rat Feces by High Performance Liquid Chromatography with Fluorescence Derivatization and Identification by Online Mass Spectrometry

    Chen Wandong, Zhang Haifeng, Guo Qingjun, Wang Xianfeng, Yourui Suo,Chen Guichen, Zhang Honghai, You Jinmao

    1College of Chemical Science, Qufu Normal University, Qufu 273165

    2College of Life Science, Qufu Normal University, Qufu 273165

    3Northwest plateau Institute of Biology, Chinese Academy of Sciences, Xining 810001)

    4Jining teachers College, Jining 272025

    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. ......

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