天冬氨酸酶体系中R-3-氨基丁酸相关物质HPLC方法研究  

Studies on HPLC method of R-3-aminobutyric acid related substances in aspartase system

作　　者：穆玉敏 韩广欣 张浩月 梁海龙 杨梦茜 刘丽荣 任丽梅 Mu Yumin;Han Guangxin;Zhang Haoyue;Liang Hailong;Yang Mengxi;Liu Lirong;Ren Limei(School of Chemical Engineering,Shijiazhuang University,Shijiazhuang 050035,China;Hebei International Joint Research Center for Biopharmaceuticals,Shijiazhuang 050035,China;Hebei Provincial University Microbial Pharmaceutical Application Technology R&D Center,Shijiazhuang 050035,China;Mei bang mei he Biological Technology Corporation Ltd.,Shijiazhuang 050000,China)

机构地区：[1]石家庄学院化工学院,河北石家庄050035 [2]河北省生物制药国际联合研究中心,河北石家庄050035 [3]河北省高效微生物制药应用技术研发中心,河北石家庄050035 [4]美邦美和生物科技有限公司,河北石家庄050000

出　　处：《煤炭与化工》2023年第5期122-125,129,共5页Coal and Chemical Industry

摘　　要：为了建立一种高效被大众认可的满足酶促体系中R-3氨基丁酸及相关物质的分析方法。基于显色法、衍生法本文探究了高效液相色谱UV检测器210 nm下采用特定流动相及检测参数的HPLC外标法。结果表明在选定的色谱条件下,R-3-氨基丁酸与相关物质可以和酶促反应中其它物质得到很好的分离,反应过程中底物巴豆酸在浓度0.05~0.4 g/L范围线性良好,R2大于3个9,产物R-3-氨基丁酸在此检测条件下灵敏度好,检测限、定量限符合规定,浓度0.35~2 g/L范围线性良好,R2大于3个9。该方法同时满足原辅料、催化反应、纯化结晶、成品含量及杂质检测。In order to establish an efficient analysis method of R-3-aminobutyric acid and related substances in the enzymatic system.Based on the color development method,the derivative method,the method of HPLC external standard at 210nm with specific mobile phase and detection parameters was explored.The results show that under selected chromatographic conditions,R-3-aminobutyric acid and related substances can be well separated from other substances in the enzymatic reaction,and the substrate crotonic acid in the concentration of 0.05~0.4 g/L Range lineargood,R2More than 999,the product R-3-aminobutyric acid has good sensitivity under this detection conditions,the detection limit and quantification limit meet the requirements,and the concentration is 0.35~2 g/L Range lineargood,R2More than 999.The method also meets the raw materials,catalytic reaction purification and crystallization,finished product content and impurity detection.

关 键 词：天冬氨酸酶突变体 生物酶催化 巴豆酸 R-3-氨基丁酸 高效液相色谱法 外标法 

分 类 号：O657.72[理学—分析化学]