Uno S, Takehiro O, Tabata R, Ozawa K
Developmental Research Department, Sanwa Kagaku Kenkyusho Do., Mie, Japan.
Clin Chem. 1995 Dec;41(12 Pt 1):1745-50.
We have developed a new, sensitive, and rapid method for measuring the ketone body concentration in arterial blood and determining the arterial blood ketone body ratio. The procedure involves the sequential use of the enzymes 3-hydroxybutyrate dehydrogenase (3-HBDH; EC 1.1.1.30) and NADH oxidase, followed by a color-generating reaction with the hydrogen peroxide produced by the oxidase reaction. The amount of oxidized chromogen produced is proportional to the 3-hydroxybutyrate (3-HBA) concentration. The acetoacetate (AcAc) concentration is obtained after complete conversion of the AcAc to 3-HBA, in the presence of 3-HBDH. The total 3-HBA concentration is measured and then subtracted from the total ketone body concentration to give the AcAc concentration. This procedure may be applied to plasma samples and the absorbance change measured with an automated chemistry analyzer. Ketone body concentration may be determined over the range 0 to 400 mumol/L. The analysis takes approximately 12 min and requires only 30 microL of plasma.
我们已经开发出一种新的、灵敏且快速的方法，用于测量动脉血中酮体浓度并测定动脉血酮体比率。该过程包括依次使用3-羟基丁酸脱氢酶（3-HBDH；EC 1.1.1.30）和NADH氧化酶，随后与氧化酶反应产生的过氧化氢进行显色反应。产生的氧化色原的量与3-羟基丁酸（3-HBA）浓度成正比。在3-HBDH存在的情况下，将乙酰乙酸（AcAc）完全转化为3-HBA后，可得到AcAc浓度。测量总3-HBA浓度，然后从总酮体浓度中减去该值，即可得到AcAc浓度。此方法可应用于血浆样本，并使用自动化学分析仪测量吸光度变化。酮体浓度的测定范围为0至400μmol/L。分析过程大约需要12分钟，仅需30μL血浆。
