Pre-column derivatization-high performance liquid chromatography for the determination of aliphatic amines with fluorescence detection and mass spectrometric identification

A simple and highly sensitive method based on the derivatization of aliphatic amines with 2-(2-phenyl-1 H-phenanthro-[9, 10-d] imidazole-1-yl)-acetic acid (PPIA) followed by high performance liquid chromatography with fluorescence detection and on-line post-column mass spectrometric identification has been developed. Optimum derivatization, giving the corresponding stable fluorescent derivatives, was obtained by reacting of aliphatic amines with PPIA at 60 degrees C for 15 min in the presence of N-ethyl-N'-(3-dimethylamino propyl) carbodiimide hydrochloride (EDC) catalyst in acetonitrile solvent. The fluorescence detection Wavelength of PPIA derivatives was 380 nm (excitation wavelength 260 nm). Separation of the derivatized amines with a good baseline resolution in conjunction with a gradient elution was carried out on a reversed phase Eclipse XDB-C-8 column (4.6 min x 150 min, 5 mu m), in this way, the content of free fatty amines in waste water of paper mill, telencephalon tissue of rat and acidophilus milk was determined. The identification of aliphatic amine derivatives was carried out by post-column online mass spectrometry with atmospheric pressure chemical ionization source under positive-ion mode detection. The established method exhibits excellent reproducibility and recovery. Excellent linear responses are observed with regression coefficients > 0.9996, detection limits ( at signal-to-noise of 3: 1) are 3.1-18 fmol.