| 金属离子增稳的酶法终点法测定1,5-脱水葡糖醇 |
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| 引用本文： | 金秀儿,陈婉君,李建珍,陈桠雯,连国军,曹建明.金属离子增稳的酶法终点法测定1,5-脱水葡糖醇[J].广东微量元素科学,2012,19(8):14-17. |
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| 作者姓名： | 金秀儿 陈婉君 李建珍 陈桠雯 连国军 曹建明 |
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| 作者单位： | 温州医学院环境与公共卫生学院,浙江温州325000 |
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| 基金项目： | 浙江省大学生科技创新活动计划（新苗人才计划,编号2011R413014） |
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| 摘 要： | 为建立金属离子增稳的酶法终点法测定1,5-脱水葡糖醇,通过葡萄糖激酶(GK)和葡萄糖-6-磷酸脱氢酶(G6PD)将样品中葡萄糖(Glu)转化为6-磷酸葡萄糖内酯(G6PL),1,5-AG经吡喃糖氧化酶(PROD)作用生成1,5-脱水果糖(1,5-AF)和H2O2,后者借Trinder’s系统显色定量。结果表明,该法线性范围0～250μmol/L,平均回收率为99.3%,批内变异系数(CV)和批间变异系数分别为0.009 8～0.027 7、0.016 2～0.036 8,与气相色谱-质谱法(GC-MS)比较具有良好的相关性,线性回归方程和相关系数分别为y=0.982 x+2.68,r=0.998 8。以金属离子为增稳剂的酶法终点法测定1,5-AG含量具有简便、灵敏、可靠的特点,适合实际推广应用。
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| 关 键 词： | 1 5-脱水葡糖醇 光度分析法 酶法终点法 |
A New Enzymatic End Point Assay of 1,5 -AG Using Metal Ions as Stabilizer |
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| Institution: | JIN Xiuer, CHEN Wanjun, LI Jianzhen, et al. ( School of Environmental Science and Public Health, Wenzhou Medical College, Wenzhou 325000, China) |
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| Abstract: | To establish a new enzymatic end point assay of 1,5 -AG using metal ions as stabilizer. By using glucokinase （GK） and glucose -6 -phosphate dehydrogenase （G6PD）, glucose （Glu） was converted to Glucose 6 - phosphate lactone （G6PL）. The 1,5 - AG was oxidized by the pyranose oxidase （PROD） and converted into 1,5 -dehydration of fructose （1,5 -AF） and I-I202, the latter was quantitatived by Trinder＇s color quantification system. The method was liner in the range of 0 250 μmol/L, the average rate of recovery was 99. 3% , the withinrun and between run CV were 0. 009 8 -0. 027 7 and 0. 016 2 -0. 036 8, respectively. Comparing this method （y） with GC -MS （x）, as the regression equation was obtained ： y = 0. 982 x ＋ 2.68, r = 0. 998 8. This method was simple, sensitive and suitable for application. |
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| Keywords: | 1 5 - AG spectrophotometry enzymatic end - point method |
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