鼠类粪便中类固醇的荧光标记及质谱鉴定  

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

作　　者：陈万东[1] 张海峰[2] 郭庆君[1] 王贤峰[3] 索有瑞[4] 陈桂琛[4] 张洪海[3] 尤进茂[2] 

机构地区：[1]济宁师范专科学校,济宁272025 [2]曲阜师范大学化学科学学院,曲阜273165 [3]曲阜师范大学生命科学学院,曲阜273165 [4]中国科学院西北高原生物研究所,西宁810001

出　　处：《分析化学》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)

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.

关 键 词：高效液相色谱-质谱 荧光检测 柱前衍生 类固醇 

分 类 号：Q548-3[生物学—生物化学]