脂肪胺荧光衍生物的高效液相色谱分离及质谱鉴定

采用新型荧光衍生试剂2-(9-吖啶酮)-乙酸(AAA)进行柱前衍生并经荧光检测对脂肪胺进行了高效液相色谱(HPLC)分离和在线质谱定性.衍生物荧光激发和发射波长为λex=404nm,λem=440nm.30℃下在乙腈溶剂中用N-乙基-N′-(3-二甲氨基)丙基]碳二亚胺盐酸盐(EDC)做催化剂,衍生反应20min后获得稳定的荧光产物.在HypersilBDSC18(4.6mm×100mm,5μm)色谱柱上,采用梯度洗脱对12种脂肪胺衍生物进行了优化分离.采用大气压化学电离源(APCISource)正离子模式进行在线柱后质谱定性,实现了各种脂肪胺衍生物的快速、准确测定.该方法具有良好的重现性,多数脂肪胺的线性回归系数大于0.9996,检测限为12.09~25.52fmol.

HPLC Determination of Aliphatic Amines with Fluorescence Detection and Mass Spectrum Identification

A simple and high sensitive method based on the derivatization of aliphatic amines with 2(9acridone)acetic acid (AAA)followed by high performance liquid chromatography(HPLC)with fluorescence detection and postcolumn mass spectrometric identification has been developed. The fluorescence excitation and emission wavelengthesof derivatives were set at λ_ ex =404and λ_ em=440 nm, respectively. Optimum derivatization,giving the corresponding stablefluorescent derivatives, was obtained by reacting of aliphatic amines with AAA at30 ℃ for 20 min in the presence of NethylN′(3dimethylamino propyl)carbodiimide hydrochloride (EDC) catalyst in acetonitrile. Separation of the derivatized amines with a good baseline resolution in conjunction with a gradient elution was achieved on a reversedphase Hypersil BDS C_ 18(4.6 mm×100 mm, 5μm)column. The identification of aliphatic aminederivatives was carriedout by postcolumn online mass spectrum with atmospheric pressure chemical ionization (APCI) source under positive ion modedetection.The establishedmethod exhibits excellentreproducibility and recovery. Excellent linear responsesareobserved with coefficients >0.999 6,detection limits (at signaltonoise of 3∶1) are12.09~ 25.52 fmol.