柱前衍生-HPLC法测定五指毛桃中16种游离氨基酸含量  

Determination of 16 Free Amino Acids of Ficus hirta by Pre-column Derivatization-High Performance Liquid Chromatography

作　　者：陈荣珠 王小平 高伟城[1] 陈育青[1] 丁其春 CHEN Rongzhu;WANG Xiaoping;GAO Weicheng;CHEN Yuqing;DING Qichun(Department of Pharmacy,Zhangzhou Health Vocational College,Zhangzhou 363000,China)

机构地区：[1]漳州卫生职业学院药学系,福建漳州363000

出　　处：《食品工业科技》2022年第17期306-315,共10页Science and Technology of Food Industry

基　　金：漳州市自然科学基金(ZZ2021J16);福建省中青年教师教育科研项目(科技类)(JAT210865);漳州市科技特派员项目(20210187);漳州卫生职业学院高学历人才专项(ZZWYGXL01);漳州卫生职业学院科技创新团队(Kjcx-5)。

摘　　要：目的:建立柱前衍生-高效液相色谱法同时测定不同产地、不同叶形五指毛桃Ficus hirta Vahl根中16种氨基酸含量,评价其质量差异。方法:采用高效液相色谱法,以异硫氰酸苯酯进行柱前衍生化,采用Ultimate Amino Acid色谱柱(4.6 mm×250 mm,5μm),以0.1 mol/L乙酸钠-乙腈为流动相,进行梯度洗脱,流速1.0 mL/min,柱温为40℃,检测波长为254 nm。结果:9个产地12份五指毛桃根中16种氨基酸线性关系良好,相关系数均大于0.9992,平均加样回收率介于98.1%~105.9%之间,RSD为0.67%~2.11%,L-天门冬氨酸、L-谷氨酸、L-丝氨酸、甘氨酸、L-组氨酸、L-精氨酸、L-苏氨酸、L-丙氨酸、L-脯氨酸、L-酪氨酸、L-缬氨酸、L-甲硫氨酸、L-异亮氨酸、L-亮氨酸、L-苯丙氨酸和L-赖氨酸的平均含量分别为3.308、3.424、2.750、2.978、1.302、4.549、2.778、3.989、3.818、1.327、3.732、0.395、2.925、4.549、2.873、3.305 mg/g。产地为漳州卫生职业学院的四种叶形五指毛桃根中氨基酸总含量由高到低为,三指浅裂>七指深裂>五指深裂>全缘;其余8个产地的五指毛桃根中16种氨基酸总含量也存在差异,含量最高的为广西钦州灵山县,其次为广东河源源城区源南镇,含量最低的为福建漳州龙海区程溪镇。结论:该方法能使16种氨基酸很好地进行分离,可靠准确,且加样回收率较高,可作为五指毛桃根中氨基酸含量的测定方法,并可以用氨基酸含量来评价五指毛桃质量。Objects:To evaluate the quality of the root of Ficus hirta Vahl,a pre-column derivation-high performance liquid chromatography(HPLC) method was established for the determination of 16 kinds of amino acids in the root of Ficus hirta from different origins and leaf shapes.Methods:The contents of 16 kinds of amino acids in Ficus hirta were determined by HPLC with phenyl isothiocyanate pre-column derivatization method.The separation was achieved using a ultimate amino acid column(4.6 mm×250 mm,5 μm) under the gradient elution with the mobile phases composed of 0.1 mol/L sodium acetate buffer and acetonitrile at the flow rate of 1.0 mL/min,column temperature of 40 ℃ and UV detector wavelength at 254 nm.Results:The contents of 16 kinds of amino acids had a good linear relationship,and the correlation coefficients was more than 0.9992,while the average recovery rates(n=6) were 98.1%~105.9% and RSD were 0.67%~2.11%.The average contents of L-asparaginic acid,L-glutamic acid,L-serine,glycine,L-histidine,L-arginine,L-threonine,L-alanine,L-proline,L-tyrosine,L-valine,L-methionine,L-isoleucine,L-leucine,L-phenylalanine,L-lysine were respectively 3.308,3.424,2.750,2.978,1.302,4.549,2.778,3.989,3.818,1.327,3.732,0.395,2.925,4.549,2.873,3.305 mg/g.There were differences in the content of total amino acid in Ficus hirta from different leaf shapes.Compared with leaf shapes,the three-finger shapet had the highest total amino acid,followed by seven-finger shpape,and then five-finger shape,and the last was undivided.The content of total amino acid in Ficus hirta from different producing areas were also different.The highest content was in Lingshan county of Qinzhou city of Guangxi province,followed by Yuannan town of Heyuan city of Guangdong province,and the lowest content was in Chengxi town of Longhai district of Zhangzhou city of Fujian province.Conclusion:16 kinds of amino acids were separated by this method,which was reliable,accurate and had high recovery rate.It could be used for the determination of amino acids in Ficus h

关 键 词：五指毛桃 氨基酸 柱前衍生化 异硫氰酸苯酯 高效液相色谱法 

分 类 号：R282.5[医药卫生—中药学] R282.6[医药卫生—中医学]