Abstract
A highly sensitive method for the determination of free fatty acids (FFAs) using high-performance liquid chromatography (HPLC) with fluorescence detection has been developed. Twenty-six FFAs (C5–C30) were derivatized by a novel pre-column derivatization reagent 2-(11H-benzo[a]carbazol-11-yl)-ethyl-4-methyl- benzenesulphonate (BCETS). Derivatives were sufficiently stable to be efficiently analyzed by HPLC. All fatty acid derivatives were separated on a hypersil BDS-C8 column in conjunction with a gradient elution with a good baseline resolution. The identification was carried out by post-column online mass spectrometry with atmospheric pressure chemical ionization (APCI) in positive-ion detection mode. The established method exhibited good selectivity and excellent repeatability. The RSD (n = 6) for each fatty acid derivative was <2.8%, and detection limit (at signal-to-noise of 3:1) was 15–64 fmol. The method was used to quantify FFAs in Cortex Lycii sample with satisfactory results.
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