胖大海中氨基酸的含量测定及多元化统计分析

Determination and Diversification of Amino Acids in Scaphium scaphigerum

目的:测定胖大海中氨基酸的含量,实现对不同产地胖大海的质量评价。方法:胖大海以6 mol/L盐酸于140℃水解2 h,以邻苯二甲醛/9-芴甲氧羰酰氯(OPA/FMOC)为衍生化试剂进行柱前衍生,用反相高效液相色谱法测定23种氨基酸的含量;对不同产地胖大海中氨基酸的含量进行综合评价。采用Agilent Advance Bio AAA色谱柱(4.6 mm×100 mm,2.7μm),以10 mmol/L磷酸氢二钠+10 mmol/L硼酸钠缓冲液(用盐酸调节pH值至8.2)为流动相A,甲醇-乙腈-水(45:45:10)为流动相B,梯度洗脱,流速1.0 mL/min,柱温40℃,检测波长338、262 nm,进样量1μL。利用聚类分析、主成分分析及偏最小二乘法对不同产地胖大海中的氨基酸进行综合评价。结果:含量测定方法学验证结果良好。通过聚类分析可将样品分为3类,结合主成分分析法、偏最小二乘法判别分析发现其中7种不同氨基酸因产地不同导致的差异。结论:研究所建立的氨基酸含量测定方法可以用于胖大海中氨基酸的定量分析,通过主成分分析法、聚类分析及偏最小二乘法可用于区分不同产地胖大海氨基酸的成分差异,为胖大海中的氨基酸评价及资源的开发提供理论基础。

The content of amino acids in Scaphium scaphigerum was determined to evaluate the quality from different habitats.Methods:6 mol/L hydrochloric acid was hydrolyzed at 140℃for 2 h in Scaphium scaphigerum,OPA/FMOC was used as the derivativator for column derivativation.The quantities of 23 amino acids were determined by reversed-phase high performance liquid chromatography.The contents of amino acids in the Scaphium scaphigerum from different habitats were evaluated synthetically.Agilent Advance Bio AAA chromatographic column(4.6 mm×100 mm,2.7μm),10 mmol/L disodium hydrogen phosphate+10 mmol/L sodium borate buffer(pH adjusted by hydrochloric acid to 8.2)as mobile phase A,methanol-acetonitrile-water(45∶45∶10)as mobile phase B,gradient elution,flow rate 1.0 mL/min,column temperature 40℃,detection wavelength 338 nm,262 nm,injection volume 1μL.Principal component analysis,cluster analysis,and partial least square method were used to evaluate the amino acids from different habitats.Results:The method of content determination proved to be good.The samples can be divided into 3 categories by cluster analysis,and the discriminant analysis of principal component analysis and partial least square method shows that 7 amino acids are different due to the difference in origin.Conclusion:The amino acid content determination method established in this study can be used for the quantitative analysis of Scaphium scaphigerum,and the principal component analysis,clustering analysis and partial least square method can be used to distinguish the amino acid composition differences of Scaphium scaphigerum from different regions,providing a theoretical basis for the amino acid evaluation and resource development in Scaphium scaphigerum.