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建立了水产品中3种游离芳香族氨基酸含量的HPLC测定方法。试验确定了流动相的最佳pH、流速及浓度,并评价了方法的准确度、精密度和检测限。采用5%高氯酸溶液提取,用KOH溶液中和后进行HPLC分析。采用WatersAtlantis T3 C18(4.6×250 mm,5μm)色谱柱分离,以15%甲醇-85%磷酸盐溶液为流动相,用磷酸调pH为6.0;色谱条件流速:0.9 mL/min;柱温:27℃;检测波长:256.7 nm;12 min内完成测定。结果表明:Tyr、Phe、Trp分别在6～500μg/mL、30～1 000μg/mL和3～250μg/mL范围内有良好的线性关系,其线性相关系数(R2)在0.998 6～0.999 5之间。三者的检测限分别为2μg/mL、10μg/mL、1μg/mL,加标回收率在88.0%～105.2%之间。方法操作简便且定性可靠、定量准确。
A HPLC method for the determination of three free aromatic amino acids in aquatic products was established. The experiment determined the optimum pH, flow rate and concentration of the mobile phase, and evaluated the accuracy, precision and detection limit of the method. Extraction with 5% perchloric acid solution, neutralization with KOH solution and HPLC analysis. The sample was separated on a Waters Atlantis T3 C18 (4.6 × 250 mm, 5 μm) column using 15% methanol-85% phosphate as the mobile phase and adjusted to pH 6.0 with phosphoric acid. The chromatographic conditions were as follows: flow rate: 0.9 mL / ℃; detection wavelength: 256.7 nm; determination within 12 min. The results showed that there was a good linear relationship between Tyr, Phe and Trp in the range of 6 ~ 500μg / mL, 30 ~ 1 000μg / mL and 3 ~ 250μg / mL respectively, and the linear correlation coefficient was 0.998 6 ~ 0.999 5 between. The detection limits of the three were 2μg / mL, 10μg / mL and 1μg / mL respectively. The recoveries of spiked samples ranged from 88.0% to 105.2%. The method is simple, reliable and quantitative.