2-(11H-苯[a]咔唑)-乙酸在脂肪胺的高效液相色谱分离及质谱鉴定中的应用

Determination of Aliphatic Amines Using Pre-column Derivatization and HPLC with Fluorescence and Mass Spectrometric Detections

采用新型荧光衍生试剂2-（11H-苯[a]咔唑）-乙酸（BCA）进行柱前衍生并经荧光检测对脂肪胺进行高效液相色谱（HPLC）分离和质谱定性。衍生物荧光激发和发射波长为λex=285nm，λem=384nm。60℃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）作催化剂，衍生反应15min后获得稳定的荧光产物。在Hypersil BDS C18（4．0mm×200mm，10μm）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（APCI Source）正离子模式进行质谱定性，实现了各种脂肪胺衍生物的快速、准确测定。脂肪胺的线性回归系数不小于0．9998，检出限（S／N=3）为5．73～31．3fmol。

A simple and highly sensitive method based on the derivatization of aliphatic amines with 2- （ 11H-benzo [ a ] carbazol-11 -yl ） -acetic acid （ BCA ） followed by high-performance liquid chromatography （HPLC） with fluorescence detection and mass spectrometric identification was developed. The fluorescence excitation and emission wavelengths of derivatives were set at λex = 285 nm and ,λem = 384 nm, respectively. A stable fluorescent derivative was obtained by reacting of with BCA at 60 ℃ for 15 min in the presence of N-ethyl-N′- （3-dimethylamino propyl） carbodiimide hydrochloride （EDC） as catalyst in acetonitrile solvent. The separation of the derivatized amines with a good baseline resolution was achieved on a reversed-phase Hypersil BDS C18（4. 0 mm ×200 mm, 10 μm） column with a gradient elution. The identifications of aliphatic amine derivatives were carried out by mass spectrometry with atmospheric pressure chemical ionization（APCI） source under positive-ion mode detection. The established method exhibited excellent reproducibility and recovery. Excellent linear responses were observed with correlation coefficients no less than 0. 999 8, detection limits（signal-to-noise ratio of 3 ： 1） of 5. 73-31.3 fmol.