一种简便、准确的血清氧化及还原型辅酶II的分光光度测定法的建立

A Simple and Reliable Spectrophotometric Method for Quantification of Pyridine Nucleotides in Serum

由于辅酶 II在正常细胞稳态及许多代谢异常中均非常重要 ,寻找一个简便而可靠的方法测定氧化型及还原型辅酶 II对于了解机体的氧化还原稳态将非常有益。本法是对 Nisselbam法稍加改进 ,运用一个敏感的酶循环反应以分光光度法测定血清中的辅酶 II,结果显示标准曲线线性关系良好 ,加样回收较完全。用本法测定的 2 7例健康献血员 ,血清 NADPH、NADP+ 及其比值 NADPH/ NADP+ 分别为 8.385± 1.5 16 nm ol/ ml,3.6 2 4± 0 .985nmol/ m l,2 .36 12± 0 .80 5 ,男女性别无明显差异。本法灵敏、准确、重现性好、费用低、操作简便 ,是测定氧化型及还原型辅酶

Pyridine nucleotides plays an important role in the maintenance of cellular homeostasis and its disturbance also involves in many perturbance of metabolism, so a simple and reliable assay for determination of oxidized and reduced pyridine nucleotides in serum would be significantly necessary in understanding the redox status of body. A novel modified method, which is based on Nisselbam and making use of a cycling enzymatic reaction and spectrophotometric assay to measure the pyridine nucleotides in serum, showed good results. The standard curve has fine linear relationship; the recovery of added standard samples is almost perfect. Tewnty seven samples from 27 normal subjects were examined by the method. The results were: NADPH 8.385±1.516 nmol/ml,NADP + 3 624±0.985 nmol/ml, and NADPH/NADP + 2.3612±0.8057;no significant difference in the variables was found between the male and female. This is a sensitive, simple, reliable and easy practicing method for oxidized and reduced pyridine nucleotides assay.