[关键词]
[摘要]
目的 分析淡豆豉Sojae Semen Preparatum和大豆Glycine max中游离及水解氨基酸。方法 用0.1 mol/L HCl作溶剂超声提取得到淡豆豉和大豆中的游离氨基酸;用6 mol/L HCl水解提取得到水解氨基酸,以异硫氰酸苯酯(PITC)为柱前衍生化试剂,采用柱前衍生RP-HPLC法进行分析。结果 淡豆豉和大豆中均含有16种氨基酸,这16种氨基酸浓度在0.031~1.750 mmol/L内呈良好的线性关系,r均大于0.997 9,平均回收率91.02%~102.04%,RSD1.01%~4.81%。结论 本方法灵敏、准确,具有良好的重复性和稳定性,16种氨基酸能在样品杂质量较多的情况下得到分离,且采用普通C18色谱柱成本低,可广泛应用于富含氨基酸样品的分析。
[Key word]
[Abstract]
Objective To analyze the free and hydrolyzed amino acids in Sojae Semen Preparatum (SSP) and Glycine max (GM) by pre-column derivatization RP-HPLC. Methods Free amino acids extracted in HCl (0.1 mol/L) by ultrasonic method and hydrolyzed amino acids extracted in HCl (6 mol/L) by hydrolysis method were both derivated by the agent phenylisothiocyanate so as to analyze them by gradient elution. Results There were 16 kinds of amino acids in both SSP and GM. The 16 kinds of amino acids had good linearity in the range of 0.031—1.750 μmol/mL with the coefficients of correlation all over 0.9979. The average recoveries were 91.02%—102.04% and the RSD values were between 1.01% and 4.81%. Conclusion The method is not only sensitive and accurate, but also has the high repeatability and stability, and the 16 kinds of amino acids could be separated in the case of the samples with more impurities. The common C18 column is widely used for the determination of many other samples riched in amino acids at lower cost.
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[基金项目]
国家自然科学基金资助项目(81173469);教育部新世纪优秀人才资助项目(NCET-09-0899)