高效液相色谱法测定琥珀酰亚胺及其酶解产物  

Analysis of Succinimide and Its Enzymatic Product by High Performance Liquid Chromatography

作　　者：钮利喜[1] 陈云霞[1] 钞建宾[2] 袁静明[1] 

机构地区：[1]山西大学生物技术研究所教育部化学生物学与分子工程重点实验室,太原030006 [2]山西大学生物技术研究所现代化学研究所,太原030006

出　　处：《分析化学》2006年第U09期148-150,共3页Chinese Journal of Analytical Chemistry

基　　金：山西省自然科学基金资助(No.20031042)

摘　　要：采用HPLC法,以Symmetry(C18柱(4·6mm×150mm,5μm),甲醇∶10mmol/L磷酸缓冲液(pH6·5)=5∶95(V/V)为流动相,在波长210nm检测条件下,使琥珀酰亚胺及其酶解产物琥珀酰胺酸得到了较好的分离。琥珀酰亚胺与琥珀酰胺酸的质量浓度与峰面积呈良好的线性关系,相关系数均在0·999以上。该方法既可用于定量分析琥珀酰亚胺和琥珀酰胺酸,又能用于测定环二酰胺酶的活性及产物转化率。检测精度可达μg水平;用NMR确认了酶分解物质的分子结构。Succinimide and its enzymatic product succinamic acid in the presence of a cyclic imidase were analyzed by high performance liquid chromatography（HPLC）. Succinimide and succinamic acid in the reaction mixture were efficiently separated and measured on a Symmetry C18 column with the mobile-phase of methanol and 10 mmol/L phosphoric buffer solution pH 6.5（5：95, V/V） as detected at 210 nm. The linear relation of peak areas indicated that their relative coefficients reach over 0.999. This method reported here, not only succinimide and succinamic acid can be quantitatively measured at microgram level, but also the activity of cyclic imidase can be assayed with good sensitivity. Meanwhile, the molecular structure of succinimide and succinamic acid in reaction mixture was also confirmed by nuclear magnetic resonance（NMR）.

关 键 词：琥珀酰亚胺 琥珀酰胺酸 环二酰胺酶 高效液相色谱 核磁共振谱 

分 类 号：O657.72[理学—分析化学]