Abstract: A rapid quantitative method of ultra performance liquid chromatography (UPLC) has been developed for the analysis of 17 amino acids in fish eggs from Acipenser schrenckii, Huso dauricus and Acipenser ruthenus. The analytes were hydrolyzed with 6.0 mol/L HCl. The extraction solution was concentrated under low pressure and neutralized with NaOH solution before derivatization with 6-aminoquinolyl-N-hydroxy-succinimidyl carbamate (AQC) as derivatization agent in borate buffer solution (pH 8.8). Gradient UPLC separation was performed on a C18 column (BEH C18, 100 mm×2.1 mm, 1.7 μm) with 30 mmol/L ammonium acetate (pH 3.5 adjusted with formic acid) and acetonitrile (containing 0.15%(v/v) formic acid and 30 mmol/L ammonium acetate) as the mobile phases at a flow rate of 0.7 mL/min. The detection was carried out with a photo-diode array (PDA) detector and the wavelength was set at 260 nm. The linear ranges were from 5.0 to 1000 μmol/L with the correlation coefficients (r2) ≥ 0.9950. The method was validated by the analysis of seven replicates. The 17 amino acid standards were spiked in fish eggs at three levels of 100, 500 and 750 μmol/L, and the average recoveries were 75.4%-107.3% with the relative standard deviations of 2.19%-12.3%. The limits of detection (LOD) for the analytes were from 0.94 μmol/L to 4.04 μmol/L. The method was successfully applied to the analysis of the 17 amino acids in fish eggs from Acipenser schrenckii, Huso dauricus and Acipenser ruthenus. The method is simple, rapid, precise and reliable.

Key words: amino acids, fish eggs, ultra performance liquid chromatography (UPLC), precolumn derivatization