新型荧光衍生试剂用于脂肪胺的反相高效液相色谱-荧光检测分析  

Determination of aliphatic amines with a sensitive reagent by HPLC with fluorescence detection and APCI/MS identification

作　　者：刘素娟[1] 尤进茂[2,3] 

机构地区：[1]牡丹江师范学院化学化工学院院,牡丹江157012 [2]曲阜师范大学化学科学学院,曲阜273165 [3]中国科学院西北高原生物研究所所,西宁810001

出　　处：《分析试验室》2010年第11期83-86,共4页Chinese Journal of Analysis Laboratory

摘　　要：利用新型荧光试剂4-(1H-菲并[9,10-d]咪唑-2-)苯甲酸(PIBA)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。激发和发射波长分别为ex=261nm,em=443nm。80℃下在吡啶溶剂中用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10min后获得稳定的荧光产物。在EclipseXDB-C8色谱柱(4.6150mm,5mm)上,梯度洗脱对12种游离脂肪胺衍生物进行了优化分离。采用大气压化学电离源(APCI)正离子模式,实现了各种脂肪胺衍生物的测定。多数脂肪胺的线性回归系数大于0.9999,检测限为10.5～53.4fmol。A simple and sensitive method based on the derivatization of aliphatic amines with 4-(1H-phenanthro[9,10-d]imidazol-2-)benzoic acid (PIBA) followed by high-performance liquid chromatography(HPLC)with fluorescence detection and post-column mass spectrometric identification has been developed.The fluorescence excitation and emission wavelengths of derivatives were set at ex=261 and em=443nm,respectively.Optimal derivatization,giving the corresponding stable fluorescent derivatives,was obtained by the reaction of aliphatic amines with PIBA at 80℃ for 10 min in the presence of 1-ethyl-3(3-dimethylaminopropyl)-carbodiimide hydrochloride catalyst in acetonitrile.Separation of the derivatized amines in conjunction with a gradient elution was achieved on a reversed-phase Eclipse XDB C8 (4.6×150mm i.d.,5m) column with a good baseline resolution.The identification of aliphatic amine derivatives was carried out by post-column online APCI/MS (atmospheric pressure chemical ionization) in positive-ion mode.Excellent linear responses are observed with correlation coefficients larger than 0.9999,and the detection limits (at signal-to-noise of 3:1) are 10.5～53.4 fmol.

关 键 词：高效液相色谱-质谱 荧光检测 柱前衍生 脂肪胺 

分 类 号：O657.72[理学—分析化学]