辛酸TSEPIP荧光衍生物制备及荧光光谱研究与分析应用

Preparation of caprylic TSEPIP derivative and its fluorescence spectra study and application for HPLC analysis

合成了荧光衍生试剂1-[2-(对甲苯磺酸酯)乙基]-2-苯基咪唑[4,5-f]9,10-菲(TSEPIP),经衍生化和固相萃取制得了TSEPIP的辛酸衍生物(TSEPIP-C8)。研究了TSEPIP-C8在包括溶剂极性、温度、卤素离子、重金属离子等不同的溶液环境因素影响下的荧光光谱性质,为高效液相色谱荧光分析检测提供了依据。在EclipseXDB-C8反相色谱柱上,在380nm检测波长下采用梯度洗脱,实现了26种脂肪酸(FFA)TSEPIP衍生物的基线分离,应用该法完成了丹参中脂肪酸的含量分析。借助柱后串联质谱APCI大气压化学电离源正离子模式,能够实现各组分的快速定性。26种脂肪酸的线性回归系数均>0.9996,检出限为3.824～47.13fmol(进样量10μL,信噪比为3:1)。

TSEPIP-C8 derivative was prepared based on the derivatization of caprylic（C8） acid using 1-[2-（p-toluenesulfonate）ethyl]-2-phenylimidazole[4,5-f]9,10-phenanthrene（TSEPIP） as the derivatization reagent by pre-column derivatization and solid phase extraction（SPE）.Fluorescence spectra properties of TSEPIP-C8,at different solvent polarity,temperature and in the presence of different halide salts and heavy metal atoms,were studied on fluorescence spectrophotometer,which provided dependence for HPLC analysis with fluorescence detection.Separation of 26 types of derivatized fatty acids（C1-C26） was carried out at the detection wavelength 380 nm with a good baseline resolution in conjunction with a gradient elution on a reversed-phase Eclipse XDB-C8 column.By this procedure,the contents of free fatty acids in Salvia miltiorrhiza B.were determined.The simple and convenient identification of fatty acid derivatives was carried out by post-column tandem mass spectrometry with atmospheric-pressure chemical ionization（APCI） source in a positive ion mode.Excellent linear responses were observed with correlation coefficients 〉0.999 6,and the detection limits were 3.824～47.13 fmol（injection volume 10 μL,at signal-to-noise of 3：1）.