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化学学报
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化学学报 ›› 2010, Vol. 68 ›› Issue (07): 653-660. 上一篇    下一篇

研究论文

胆汁酸的双敏感探针苯并[b]吖啶酮-5-乙基对甲苯磺酸酯: 荧光检测和APCI-MS鉴定

付艳艳1，宋翠华1，秦雪芹1，李肖燕1，丁明州1，孙志伟2,3，夏莲1,2,3，索有瑞2，李玉林2，尤进茂*,1,2   

  1. (1生命有机分析重点实验室 曲阜师范大学化学科学学院 曲阜 273165)
    (2中国科学院西北高原生物研究所 西宁 810001)
    (3中国科学院研究生院 北京 100049)
  • 投稿日期:2009-09-05 修回日期:2009-10-25 发布日期:2010-04-14
  • 通讯作者: 尤进茂
  • 基金资助:

    基金项目：国家自然科学基金(批准号: 20075016)；中国科学院“百人计划”项目(N o. 328)资助

Dual-sensitive Probe 2-(12-Oxobenzo[b]acridin-5(12H)-yl)ethyl 4-Toluenesulfonate for Bile Acids: Fluorescence Detection and APCI-MS Identification

Fu Yanyan1 Song Cuihua1 Qin Xueqin1 Li Xiaoyan1 Ding Mingzhou1 Sun Zhiwei2,3 Xia Lian1,2,3 Suo Yourui2 Li Yulin2 You Jinmao*,1,2   

  1. (1 Key Laboratory of Life-organic Analysis, Qufu Normal University, Qufu 273165)
    (2 Northwest Plateau Institute of Biology, Chinese Academy of Sciences, Xining 810001)
    (3 Graduate University of Chinese Academy of Sciences, Beijing 100049)
  • Received:2009-09-05 Revised:2009-10-25 Published:2010-04-14

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在Hypersil C18色谱柱上, 利用新型荧光试剂苯并[b]吖啶酮-5-乙基对甲苯磺酸酯(BAETS)作柱前衍生化试剂, 采用梯度洗脱对10种胆汁酸衍生物进行了优化分离. 在二甲亚砜溶剂中, 以碳酸钾作催化剂, 95 ℃, 45 min后获得稳定的荧光产物. 衍生物在稳态荧光条件下, 在乙腈和甲醇水溶液中的百分离子化δ值在0%～88.83%和0%～89.15%范围内. 最大激发和发射波长为λex/λem＝272/505 nm. 采用大气压化学电离源(APCI)正离子模式实现了胆汁中胆汁酸的定性定量测定. 线性回归系数均在0.9995以上, 线性范围宽, 检出限为0.76～1.62 ng/mL.

关键词: 液相色谱-质谱, 百分离子化值, 苯并[b]吖啶酮-5-乙基对甲苯磺酸酯(BAETS), 胆汁酸

On a reversed-phased Hypersil C18 column, 10 bile acid derivatives were separated using 2-(12-oxobenzo[b]acridin-5(12H)-yl)ethyl 4-toluenesulfonate (BAETS) as pre-column derivatization reagent by high-performance liquid chromatography with fluorescence detection and mass spectrum identification in conjunction with a gradient elution. Optimum derivatization, giving the corresponding sensitively fluorescent derivatives, was obtained by the reacting of bile acids with BAETS at 95 ℃ for 30 min in the presence of basic catalyst potassium carbonate in dimethylsulfoxide solvent. The optimum excitation and emission wavelengths were at λex/λem＝272 nm/505 nm. The percent ionization δ values in aqueous acetonitrile and aqueous methanol were in the range of 0%～88.83% and 0%～89.15% respectively. The identification of bile acid derivatives was obtained on the basis of an atmosphere-pressure chemical ionization source at positive model detection. Correlation coefficients are ＞0.9995, and detection limits at signal-to-noise of 3 are 0.76～1.62 ng/mL for the labeled bile acids.

Key words: high performance liquid chromatography/ion-trap mass spectrometry, percent ionization, 2-(12-oxobenzo[b]acridin-5(12H)-yl)ethyl 4-toluenesulfonate (BAETS), bile acid