MacGregor L C, Matschinsky F M
Anal Biochem. 1984 Sep;141(2):382-9. doi: 10.1016/0003-2697(84)90058-7.
An enzymatic assay for myo-inositol (MI) is described. The method is based on the oxidation of MI by NAD+-dependent myo-inositol dehydrogenase, coupled to reoxidation of NADH with oxalacetate and malate dehydrogenase. The resultant malate is measured fluorimetrically. Several variations of the assay are described for measuring MI in serum and in tissues in amounts ranging from 0.2 pmol to 8 nmol. Highest sensitivity is achieved by applying an oil-well technique for handling small droplets, and by using the principle of enzymatic cycling. The potential of the technique is illustrated by MI measurements in several tissues of normal and diabetic rats and Chinese hamsters.
本文描述了一种用于测定肌醇(MI)的酶促测定法。该方法基于NAD⁺依赖性肌醇脱氢酶对MI的氧化作用,并与草酰乙酸和苹果酸脱氢酶将NADH再氧化相偶联。通过荧光法测定生成的苹果酸。文中描述了该测定法的几种变体,用于测定血清和组织中含量范围为0.2皮摩尔至8纳摩尔的MI。通过应用油井技术处理小液滴以及使用酶循环原理可实现最高灵敏度。通过对正常和糖尿病大鼠及中国仓鼠的多个组织进行MI测量,展示了该技术的潜力。