反相高效液相色谱法测定白屈菜种子中挥发油 成分分析及氨基酸含量

Determination of amino acids and analysis of the volatile oil in seeds of Che-lidonium majus by reversed phase high-performance liquid chromatography

中文关键词:  白屈菜种子  挥发油  气质联用  氨基酸  反相高效液相色谱法  异硫氰酸苯酯

英文关键词:the seeds of Chelidonium majus  volatile oil  gas chromatography-mass spectrometry (GC-MS)  Amino acids  phenyl isothiocyanate  pre-column derivation  RP-HPLC

中文摘要:

      目的 研究白屈菜种子中挥发油的化学成分并建立一种反相高效液相色谱法对白屈菜种子中游离氨基酸和水解氨基酸含量进行测定。方法 采用气质联用的方法对白屈菜种子中的挥发油成分进行分析, 采用反相高效液相色谱法, 以异硫氰酸苯酯为衍生试剂进行柱前衍生, 采用梯度洗脱的方法, 检测白屈菜种子中游离氨基酸和水解氨基酸含量。结果 挥发油中共鉴定出化学成分27个, 其中亚油酸的含量最高可达到65%左右。氨基酸的检测浓度线性范围为0.0020~0.2700 mmol/L, r均≥0.9991。21种游离氨基酸的平均加样回收率在87.88%~100.10%之间, RSD在0.62%~2.58%之间(n=6); 水解氨基酸的平均加样回收率在91.79%~100.03% 之间, RSD在0.48%~1.97% 之间(n=6)。结论 白屈菜种子种中挥发油的主要成分为不饱和脂肪酸, 本文建立的氨基酸测定方法经方法学验证适合白屈菜种子中游离氨基酸和水解氨基酸的含量测定。

英文摘要:

      Objective To investigate the volatile oil in seeds of Chelidonium majus and determinate the content of 21 free and hydrolyzed amino acids in the seeds of Chelidonium majus by pre-column derivatization reversed phase high-performance liquid chromatography (RP-HPLC). Methods The volatile oils in samples were identified and determined by gas chromatography-mass spectrometry. The samples were derived with phenyl isothiocyanate and detected by RP-HPLC. Free and hydrolyzed amino acids content in seeds of Chelidonium majus was determined by gradient elution chromatography. Results Twenty-seven compounds were separated in volatile oil, and the content of linoleic acid could reach about 65%. There was a good linear relationship of the 21 amino acids in the range of 0.0020~0.2700 mmol/L (the coefficients of correlation were all over 0.9991). The average recoveries of free amino acids were 87.88%~100.10% (RSD=0.62%~2.58%, n=6); the average recoveries of hydrolyzed amino acids were 91.79%~100.03% (RSD=0.48%~1.97%, n=6). Conclusion The main component in the volatile oil in seed of Chelidonium majus is unsaturated fatty acid. The methodology validation shows that the method is suitable for determination of free and hydrolyzed amino acids in Chelidonium majus.