CAJ | 학술논문

采用溶胶凝胶技术分别固定了胆固醇脂酶和胆固醇氧化酶,制成固定化酶柱;人体血清中胆固醇脂在胆固醇脂酶的催化作用下生成胆固醇,胆固醇在胆固醇氧化酶的催化作用下被氧化产生H_2O_2,将其与鲁米诺发生耦合的化学发光反应,在模拟酶血红蛋白的催化作用下产生较强的化学发光.结合流动注射技术,建立了溶胶凝胶固定化酶流动注射化学发光法测定胆固醇的新方法.实验发现,发光强度与胆固醇的浓度在一定范围内呈良好的线性关系,总胆固醇的线性范围为1.01×10~(- 6)～2.02×10~(-4) mol/L(r=0.9975);检出限为7.5×10~(- 7) mol/L;游离胆固醇的线性范围为5.0×10~(- 8)～2.18×10~(-5 )mol/L(r=0.9991);检出限为5.0×10~(-9) mol/L.用生化分析仪(东芝TBA-120FR)与本方法进行对照,两种方法无显著性差异.本方法已应用于临床血清样品中胆固醇的检测.
채용용효응효기술분별고정료담고순지매화담고순양화매,제성고정화매주;인체혈청중담고순지재담고순지매적최화작용하생성담고순,담고순재담고순양화매적최화작용하피양화산생H_2O_2,장기여로미낙발생우합적화학발광반응,재모의매혈홍단백적최화작용하산생교강적화학발광.결합류동주사기술,건립료용효응효고정화매류동주사화학발광법측정담고순적신방법.실험발현,발광강도여담고순적농도재일정범위내정량호적선성관계,총담고순적선성범위위1.01×10~(- 6)～2.02×10~(-4) mol/L(r=0.9975);검출한위7.5×10~(- 7) mol/L;유리담고순적선성범위위5.0×10~(- 8)～2.18×10~(-5 )mol/L(r=0.9991);검출한위5.0×10~(-9) mol/L.용생화분석의(동지TBA-120FR)여본방법진행대조,량충방법무현저성차이.본방법이응용우림상혈청양품중담고순적검측.
The cholesterol esterase and its oxidase were respectively immobilized onto sol-gel(tetraethyl orthosilicate)and then an enzymatic reaction column was prepared. Hydrolysis of cholesterol ester took place in the presence of cholesterol esterase to form cholesterol under catalysis of cholesterol oxidase, and Cholesterol was oxidized to produce hydrogen peroxide which reacted with luminal in the presence of simulated enzyme hemoglobin to result in the emission of light. Thereupon, a chemiluminescence system combined with flow injection technology by sol-gel immobilized enzyme was developed for the determination of cholesterol including free and total amount in human serum. The CL intensity was well linear with the concentration of cholesterol;liner range of total and free cholesterol was 1.01×10~(-6)～2.02×10~(-4) mol/L with a detection limit of 7.5×10~(-7) mol/L) and 5.0×10~(-8)～2.18×10~(-5) mol/L) with a detection limit of 5.0×10~(-9) mol/L respectively. Contrasted the method) with the biochemical analyzer (TBA-120FR), both the methods had no significant difference. This method has been successfully applied for the detection of cholesterol in clinic serum samples.