邻苯二甲醛柱前衍生化法快速高效测定液体生物样品L-蛋氨酸含量  

Fast and Convenient Method to Determine The L-methionine Content in Liquid Biological Samples with O-Phthaldialdehyde（OPA）Pre-column Derivation

作　　者：霍文艳[1] 张钰文 徐天勇[2] 吴怡[2] 田长富[1] 张华堂[1] 

机构地区：[1]昆明医科大学生物医学工程研究中心 [2]昆明医科大学科研实验中心,云南昆明650500

出　　处：《昆明医科大学学报》2014年第6期12-15,共4页Journal of Kunming Medical University

基　　金：国家自然科学基金资助项目(30760287);云南省科技计划基金资助项目(2009CC022);云南省孔祥复院士工作站资助项目

摘　　要：目的 拟建立一种快速、简单、灵敏的血浆等生物样品中L-蛋氨酸的含量测定方法.方法 用邻苯二甲醛（OPA）柱前衍生化法分离并定量C57小鼠血浆中L-蛋氨酸,采用反向色谱柱ODS （4.6 mm×250 mm）;pH 7.2 10 mmol/L NaH2PO4-Na2HPO4（PB）含0.2％ THF和PB-甲醇-乙腈（50：35：15）为流动相梯度洗脱;紫外检测器340 nm检测.结果 邻苯二甲醛柱前衍生化法的最低检出限浓度为0.5 μmol/L,8 min内分离测定了血浆L-蛋氨酸的含量,12 min内血浆所有氨基酸全部洗脱;在0.5-100 μmol/L进样浓度范围内,L-蛋氨酸的峰面积与浓度呈现良好的线性关系,相关系数0.999 8,对于L-蛋氨酸的标准加入回收率在94.63％和97.99％之间.结论 邻苯二甲醛柱前衍生化法是一种快速简便的紫外检测液体生物样品中L-蛋氨酸含量的方法.Objective To establish a rapid,simple,sensitive method to determine L-methionine content in biological samples,such as blood plasma.Method The O-Phthaldialdehyde （OPA） Pre-column Derivation method was used to separate and quantify L-methionine in plasma of C57 mice,with reverse chromatographic column ODS （4.6 × 250 mm）.The mobile phase of PH 7.2 10 mmol/L NaH2PO4-Na2HPO4 （PB） containing 0.2％ THF and PB-methanol-acetonitrile （50：35：15） was used to gradiently elute; the results were tested under 340 nm with UV detector.Results The concentration of the minimum detection limit was 0.5 μmol/L.Separation and determine of the content of plasma L-methionine was finished within 8 minutes,all others amino acids in plasma were finished within 12 minutes.Between the range of 0.5-100 μmol/L sample concentration,peak area and concentration of L-methionine showed perfect linear relationship,the correlation coefficient was 0.9998.The recovery rate of the L-methionine was between 94.63％ and 97.99％.Conclusion This study established a quick and easy ultraviolet detection method for liquid L-methionine content in biological samples.

关 键 词：蛋氨酸 邻苯二甲醛 OPA柱前衍生 紫外检测 

分 类 号：Q782[生物学—分子生物学]