雷丸中16种氨基酸的柱前衍生化RP-HPLC法含量测定

目的: 测定雷丸中16种氨基酸的含量。 方法: 样品以6 mol·L^-1盐酸于150 ℃水解1 h,以异硫氰酸苯酯(PITC)为衍生化试剂衍生后进行HPLC分析。采用Phenomenex Luna C₁₈(250 mm×4.6 mm,5 μm)色谱柱,柱温为43 ℃;流动相A为0.1 mol·L^-1醋酸钠缓冲液(以醋酸调pH 6.5)-乙腈(93∶ 7),流动相B为乙腈-水(4∶ 1),梯度洗脱,流速为1.0 mL·min^-1;检测波长为254 nm。 结果: 16种氨基酸浓度在1.16~47.32 μg·mL^-1范围内呈良好的线性关系(r=0.9998~1.000);平均回收率在93.3%~107.2%之间;RSD均小于3.0%。 结论: 本方法灵敏、准确,具有良好的重复性和稳定性,可用于雷丸中氨基酸的检测。

Objective: To determine and analyze 16 amino acids in Omphalia lapidecsens Schroet. Methods: The sample was hydrolyzed for 1 hour at 150 ℃,and derivatized with phenyl isothiocyanate(PITC).HPLC was performed on a Phenomenex C₁₈(4.6 mm×250 mm,5 μm)column with gradient elution of 0.1 mol·L^-1 sodium acetate buffer solution(adjusted to pH 6.5 with acetic acid)-acetonitrile(93∶ 7)(A)and acetonitrile-water(4∶ 1)(B)at the flow rate of 1.0 mL·min^-1,detected at 254 nm.Column temperature was 43 ℃. Results: 16 amino acids had good linearity in the ranges of 1.16-47.32 μg·mL^-1(r=0.9998-1.000).The average recoveries were 93.3%-107.2%,and RSDs were less than 3.0%. Conclusion: The method is sensitive and accurate with good stability,which is useful for the determination of amino acids in Omphalia lapidecsens Schroet.