HPLC检测欧李和欧李酒中的氨基酸

Detection of Amino Acids Content in Prunus humilis Bunge and Prunus humilis Bunge Wine by HPLC

建立HPLC检测欧李果和欧李酒中氨基酸含量的方法，为研究欧李果深加工和欧李酒发酵过程中含氮类物质的变化奠定基础。以2,4-二硝基氟苯（DNFB）为柱前衍生剂，C18色谱柱（250 mmx4.6 mm，5μm），乙酸钠∶乙腈-水溶液为流动相，流速1.0 mL/min进行梯度洗脱，反相高效液相色谱，二极管阵列检测器（PDA）测定22种氨基酸标品。结果表明，所建立方法的检测相关系数R2=0.9903 1、RSD值在1.054% 2.764%之间、最低检出限LOD值为0.05 2.95μg/mL，欧李果汁中加标回收率为81.7% 110.6%，欧李酒中的回收率为78.9% 101.5%。欧李果和欧李酒中均检出18种氨基酸，包括7种人体必需氨基酸。欧李果中氨基酸总量为2726 mg/100 g，以缬氨酸、异亮氨酸和亮氨酸含量较高，分别为261.0 mg/100 g、249.5 mg/100 g和242.5 mg/100 g。欧李酒中氨基酸总量为3360.36 g/L，经发酵后脯氨酸和谷氨酰胺含量升高，分别占63.59%和11.53%。所建方法操作简便易行，生成的衍生物稳定、重现性较好，衍生后剩余的衍生化试剂对氨基酸的测定无干扰，可实现对欧李果和欧李酒中多种氨基酸的同时和快速检测。

A method for the detection of amino acids content in Prunus humilis Bunge and Prunus humilis Bunge wine by HPLC was estab-lished, which laid the foundation for the research on deep-processing of Prunus humilis Bunge and the change of nitrogen-containing substanc-es in the fermentation process. The samples were derivated by 2,4-2 nitrochlorobenzene, then separated on an C18 column （250 mmx4.6 mm, 5μm） with using a mobile phase of sodium acetate and acetonitrile-water and the flowing rate was 1.0 mL/min with gradient elution. Then 22 amino acids were detected by HPLC with the diode array detector. The correlation coefficient （R2）,precision （RSD）,limit of detection （LOD） of this method were 0.9903 % 1,1.054 % 2.764 %,0.05 2.95 μg/ mL, and 81.7 % 110.6 % respectively.The recoveries were 81.7 % 110.6%and 78.9% 101.5%in Prunus humilis Bunge juice and in Prunus humilis Bunge wine respectively.Eighteen amino acids were de-tected in both Prunus humilis Bunge and Prunus humilis Bunge wine including seven essential amino acids for human body.The total content of amino acids in Prunus humilis Bunge was 2726 mg/100 g, and the content of some amino acids were higher such as valine 261.0 mg/100 g, leucine 249.5 mg/100 g and isoleucine 242.5 mg/100 g.The total content of amino acids in wine was 3360.36 g/L,however,the content of pro-line and glutamine increased to 63.59%and 11.53%after the fermentation.Such method was simple,stable and reproducible.The rest of the re-agent had no influence on the detection after amino acids were derivated.The method could be applied to detect amino acids in Prunus humilis Bunge and Prunus humilis Bunge wine simultaneously and rapidly.