Kim H R, Rho H W, Park J W, Park B H, Kim J S, Lee M W
Department of Biochemistry, Chonbuk National University Medical School, Republic of Korea.
Anal Biochem. 1994 Dec;223(2):205-7. doi: 10.1006/abio.1994.1574.
We developed an assay system for ornithine aminotransferase (EC 2.6.1.13) using ninhydrin. Pyrroline 5-carboxylate, a product of enzymatic transamination, reacts with ninhydrin under hot acidic conditions to form a reddish pigment soluble in ethanol. The millimolar extinction coefficient of reaction product dissolved in ethanol was 16.5 at 510 nm. Acidification with perchloric acid effectively abolished the interfering color development by L-ornithine and L-glutamate. The paired activity measurement in mouse tissues by ninhydrin and o-aminobenzaldehyde methods showed a good correlation (gamma = 0.985). In our ninhydrin method, stable ninhydrin replaced unstable o-aminobenzaldehyde, and sensitivity was much higher than that with the conventional o-aminobenzaldehyde method.
我们开发了一种使用茚三酮的鸟氨酸转氨酶（EC 2.6.1.13）检测系统。酶促转氨作用的产物吡咯啉5-羧酸在热酸性条件下与茚三酮反应，形成可溶于乙醇的红色色素。溶解在乙醇中的反应产物在510 nm处的毫摩尔消光系数为16.5。用高氯酸酸化有效地消除了L-鸟氨酸和L-谷氨酸的干扰显色。通过茚三酮和邻氨基苯甲醛方法对小鼠组织进行的配对活性测量显示出良好的相关性（γ = 0.985）。在我们的茚三酮方法中，稳定的茚三酮取代了不稳定的邻氨基苯甲醛，并且灵敏度远高于传统的邻氨基苯甲醛方法。
