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DNFB柱前衍生化RP-HPLC法测定氨肽素的氨基酸含量
引用本文:王棘,战祥友,滕艳坤,张德志. DNFB柱前衍生化RP-HPLC法测定氨肽素的氨基酸含量[J]. 沈阳药科大学学报, 2003, 20(6): 428-430
作者姓名:王棘  战祥友  滕艳坤  张德志
作者单位:1. 辽宁省药品检验所生化室 辽宁 沈阳 110023
2. 商丘绿源药业有限公司 河南 商丘 476100
摘    要:目的用二硝基氟苯 (DNFB)柱前衍生化RP HPLC法测定氨肽素中氨基酸的含量。方法采用二硝基氟苯为衍生试剂进行衍生 ,ODS(KromasilC18,4 6mm× 2 5 0mm ,5 μm)色谱柱分离 ,以乙腈 0 0 5mol·L-1乙酸钠水溶液为流动相 ,梯度洗脱 ,3 60nm检测 ,在 3 0min内 ,测定氨肽素的氨基酸组成与含量。结果线性范围 (总氨基酸 1 4种 )为 0 1 5~ 0 75 g·L-1;回收率为 94 8%~ 1 0 3 5 % ;RSD为0 2 6%~ 1 47% (n =5 )。结论DNFB柱前衍生化RP HPLC法可有效控制产品质量

关 键 词:高效液相色谱法  氨基酸分析  柱前衍生化法
文章编号:1006-2858(2003)06-0428-03
修稿时间:2003-02-24

Determination of the content of amino acid by pre-column derivatization and RP-HPLC
WANG Ji ,ZHAN Xiang-you ,TENG Yan-kun ,ZHANG De-zhi. Determination of the content of amino acid by pre-column derivatization and RP-HPLC[J]. Journal of Shenyang Pharmaceutical University, 2003, 20(6): 428-430
Authors:WANG Ji   ZHAN Xiang-you   TENG Yan-kun   ZHANG De-zhi
Affiliation:WANG Ji 1,ZHAN Xiang-you 1,TENG Yan-kun 1,ZHANG De-zhi 2
Abstract:Objective To develop a pre-column derivatization procedure for the determination of the content of amino acid in the sample by reversed-phase high performance liquid chromatography.Method DNFB was used as a derivatization reagent and separation of DNP-amino acid derivatives were performed on an ODS column. The signals were detected at 360 nm with the UV detector. Result Under the obove conditions all the 14 amino acids in the standard solutions were separated to baseline in 30 min.Conclusion The method is applied to analyzing amino acid composition of ampeptide and good results are obtained.
Keywords:HPLC  amino acid analysis  pre-column derivatization  
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