Rapid analysis of free aromatic amino acids in aquatic product by high performance liquid chromatography

To establish a HPLC method for the determination of free aromatic amino acids in aquatic product.Optimal conditions such as pH,flow rate and concentration of mobile phase were confirmed.Accuracy,precision and limit of quantitation were surveyed and evaluated.The method is based on an extraction of amino acids from aquatic product with a solution of 5% perchloric acid followed by neutralization by the addition of KOH just prior to HPLC analysis.HPLC system consisted of a Waters Atlantis T3 C18(4.6 × 250 mm,5 μm) column.The mobile phase consisted of 85% 0.05 mol / L sodium phosphate buffer and 15% methanol,adjusted to pH 6.0 with phosphoric acid.The flow rate was 0.9 mL / min,the column temperature was 27 ℃,and the detection wavelength was 256.7 nm.The three amino acids were separated in 12 min.The standard curves of Tyr,Phe,Trp were linear within a range of 10-500,50-1 000 and 5-250 μg / mL,and the correlation coefficient deviations range from 0.999 86 to 0.995.The limits of detection were 2,10 and 1 μg / mL.The average recovery of spiked sample deviations ranges from 88.0% to 105.2%,In conclusion,this method proved to be simple,rapid,reliable and with high accuracy and precision.It has practical value for detecting the three amino acids in aquatic product.