柱前衍生-HPLC法同时测定不同产地茯苓中18种氨基酸含量  

Pre-column derivation HPLC determination of 18 amino acids in Poria from different areas

作　　者：陈蓉 张超 顾倩 张华锋 顾炳仁 

机构地区：[1]苏州市食品药品检验所,苏州215104

出　　处：《药物分析杂志》2017年第2期297-303,共7页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立柱前衍生-高效液相色谱法测定茯苓中18种氨基酸的含量。方法:以6 mol·L^(-1)盐酸150℃水解1 h,以异硫氰酸苯酯为柱前衍生化试剂衍生后进样分析。采用C_(18) 110A色谱柱(250 mm×4.6 mm,5μm),柱温43℃,流动相A为0.1 mol·L^(-1)醋酸钠缓冲液(以醋酸调p H 6.5)-乙腈(93∶7),流动相B为乙腈-水(4∶1),进行梯度洗脱,流速1.0 m L·min^(-1),检测波长254 nm。结果:天冬氨酸(Asp)、谷氨酸(Glu)、丝氨酸(Ser)、甘氨酸(Gly)、组氨酸(His)、精氨酸(Arg)、苏氨酸(Thr)、丙氨酸(Ala)、脯氨酸(Pro)、酪氨酸(Tyr)、缬氨酸(Val)、蛋氨酸(Met)、异亮氨酸(Ile)、亮氨酸(Leu)、苯丙氨酸(Phe)、赖氨酸(Lys)、胱氨酸(Cys)、色氨酸(Trp)共18种氨基酸质量浓度在5.889~119.0μg·m L^(-1)范围内呈良好线性关系(r=0.999 9~1.000);平均回收率在93.3%~106.2%之间,RSD均小于3.0%。各产地均以脯氨酸、蛋氨酸和丝氨酸含量为最高;聚类分析将12个产地分为三类。结论:本方法可用于茯苓药材的质量控制。Objective：To establish a method for the determination of 18 amino acids in Poria from different areas by pre-column derivation HPLC.Methods：The sample was hydrolyzed with 6 mol·L（-1） hydrochloric acid for 1 hour at 150 ℃,and derivatized with phenyl isothiocyanate（PITC）.HPLC was performed on a Phenomenex Gemini C18 110A（250 mm×4.6 mm,5 μm）column with gradient elution of 0.1 mol·L（-1） sodium acetate buffer（adjusted to p H 6.5 with acetic acid）-acetonitrile（93∶7）（A）and acetonitrile-water（4∶1）（B）at the flow rate of 1.0 m L·min^-1,and the detection wavelength was 254 nm.The column temperature was 43 ℃.Result：The 18 amino acids had good linearity in the range of 5.889（-1）19.0 μg·m L^-1（r=0.999 9（-1）.000）,including aspartic acid（Asp）,glutamic acid（Glu）,serine（Ser）,glycine（Gly）,histidine（His）,arginine（Arg）,threonine（Thr）,alanine（Ala）,proline（Pro）,tyrosine（Tyr）,valine（Val）,methionine（Met）,isoleucine（Ile）,leucine（Leu）,phenylalanine（Phe）,lysine（Lys）,cystine（Cys）,tryptophan（Trp）.The average recoveries were 93.3%（-1）06.2%,and the RSDs were less than 3.0%.Pro,Met,and Ser showed the highest amount in samples from all areas.Samples were classified into three kinds by cluster analysis.Conclusion：The method can be used for the quality control of Poria.

关 键 词：茯苓 柱前衍生化 氨基酸分布 异硫氰酸苯酯 聚类分析 药材质量评价 高效液相色谱法 

分 类 号：R917[医药卫生—药物分析学]