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A sensitive method for the determination of 11 kinds of long-chain fatty acids (LCFAs) using a new synthesized labeling reagent 2-(12-oxobenzo[b]acridin-5(12H)-yl)ethyl-p-toluenesulfonate (BAETS) by HPLC-fluorescence detection has been developed.Rapid identification of LCFA derivatives was carried out by on-line mass spectrometry with atmospheric pressure chemical ionization (APCI) source under positive ion mode.