脂肪胺化合物的HPLC荧光法测定及质谱鉴定  

Pre-column Derivatization High Performance Liquid Chromatography with Fluorescence Detection-based Quantitation and Mass Spectrometric Identification of Aliphatic Amines

作　　者：刘素娟[1] 尤进茂[2] 

机构地区：[1]牡丹江师范学院化学系,黑龙江牡丹江157012 [2]曲阜师范大学化学科学学院,山东曲阜273165

出　　处：《食品科学》2009年第20期406-410,共5页Food Science

基　　金：国家自然科学基金项目(20075016);山东省自然科学基金项目(02BS046)

摘　　要：采用合成的荧光试剂4-(1-甲基-1H-菲并[9,10-d]咪唑-2-)苯甲酸(MPIBA)对脂肪胺进行柱前衍生后,采用高效液相色谱(HPLC)分离和荧光检测。衍生物的荧光激发和发射波长分别为λex=260nm、λem=446nm。80℃条件下在四氢呋喃(THF)溶剂中用N-乙基-N'-[(3-二甲氨基)丙基]碳二亚胺盐酸盐(EDC)作催化剂,衍生反应10min后获得稳定的荧光产物。在EclipseXDB-C8色谱柱(4.6×150mm,5μm)上,采用梯度洗脱对12种游离脂肪胺衍生物进行优化分离。采用大气压化学电离源(APCI)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,检测限在10.5～53.4fmol范围内,对实际样品臭豆腐中脂肪胺的测定结果满意。In order to determine aliphatic amines, a fluorescent material, 4-（1 -methy- 1H-phenanthro [9,10-d]imidazol-2-） benzoic acid （MPIBA）, was synthesized through the derivatization of aliphatic amines at 80℃for 10 rain in the presence of 1- ethyl-3（3-dimethylaminopropyl）-carbodiimide hydrochloride （EDC · HC1） catalyst in tetrahydrofuran solution. The maximum excitation and emission wavelengths of this stable derivative were 260 and 446 nm, respectively. The separation of these derivatized amines was achieved on a reversed-phase Eclipse XDB Cs （4.6 × 150 mm i.d., 5 μ m） column using a gradient elution and exhibited a good baseline resolution. The identification of aliphatic amine derivatives was carried out by post-column online APCI/MS （atmospheric pressure chemical ionization） in a positive-ion mode. Excellent linear responses were observed with a coefficient of larger than 0.9996 and a detection limit （at signal-to-noise of 3：1） of 10.5 - 53.4 fmol. This established method provided satisfactory results for determining amines from bean curd with odor.

关 键 词：高效液相色谱 荧光检测 柱前衍生 脂肪胺 

分 类 号：TS207.3[轻工技术与工程—食品科学]