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引用本文:诸敏,黄明法.茚三酮柱后衍生法测定丙氨酰-谷氨酰胺的含量[J].中国现代应用药学,2011,28(7):669-670.
ZHU Min,HUANG Mingfa.Determination of N(2)-L-alanyl-L-glutamine by Post-Column Derivatization with Ninhydrin[J].Chin J Mod Appl Pharm(中国现代应用药学),2011,28(7):669-670.
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茚三酮柱后衍生法测定丙氨酰-谷氨酰胺的含量
诸敏1, 黄明法2
1.杭州师范大学附属医院,杭州 310015;2.浙江省食品药品检验所,杭州 310015
摘要:
目的 建立茚三酮柱后衍生化氨基酸分析仪测定N(2)-L-丙氨酰-L-谷氨酰胺(L-Ala-L-Glu)含量的方法。方法 根据L-Ala-L-Glu与茚三酮反应后溶液在570 nm处有最大吸收的原理,以钾钠缓冲液为流动相,将L-Ala-L-Glu洗脱后,与茚三酮进行柱后衍生反应,生成可以被分光光度计检测的有色物质,氨基酸分析仪测定L-Ala-L-Glu的含量。结果 L-Ala-L-Glu在1~20 mg·L-1内线性关系良好(r=0.997 9),低、中、高3种浓度的回收率分别为94.5%,97.6%和98.7%;RSD分别为0.52%,0.10%和0.06%。结论 本法操作简单,分离度良好,结果准确可靠,可用于L-Ala-L-Glu的含量测定。
关键词:  N(2)-L-丙氨酰-L-谷氨酰胺  茚三酮  柱后衍生法
DOI:
分类号:
基金项目:浙江省科技厅项目(2008C30027)
Determination of N(2)-L-alanyl-L-glutamine by Post-Column Derivatization with Ninhydrin
ZHU Min1, HUANG Mingfa2
1.Affiliated Hospital of Hangzhou Normal University,Hangzhou 310015, China;2.Zhejiang Province Institute of Drug Control, Hangzhou 310015, China
Abstract:
OBJECTIVE To establish a method using post-column derivatization with ninhydrin for determination of N(2)-L-alanyl-L-glutamine(L-Ala-L-Glu). METHODS According to principle of solution having maximum absorption in 570 nm, potassium sodium buffer as mobile phase, L-Ala-L-Glu was eluted and reacted with ninhydrin, the production can be detected by spectrophotometer, amino acid analyzer determined the content of L-Ala-L-Glu. RESULTS The linear relation was good in the range of 1-20 mg·L-1 (r=0.997 9). The recoveries of low, middle and high concentration were 94.5%, 97.6% and 98.7% with the RSD of 0.52%, 0.10% and 0.06%, respectively. CONCLUSION This method is simple, separating degree is good, results are accurate and reliable, and can be used in the determination of L-Ala-L-Glu.
Key words:  N(2)-L-alanyl-L-glutamine  ninhydrin  post-column derivatization
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