Slaughter M R, O'Brien P J
Safety Assessment, SmithKline Beecham Pharmaceuticals, The Frythe, Welwyn, Herts AL6 9AR, UK.
Clin Biochem. 2000 Oct;33(7):525-34. doi: 10.1016/s0009-9120(00)00158-2.
Develop fully automated assay of antioxidant catalatic activity of catalase.
The assay is based on standard, clinical chemistry automated analyzer methods for measuring hydrogen peroxide by using the Trinder reagent. Catalase competes with 324 U/L horseradish peroxidase (type XII) and Trinder reagent for hydrogen peroxide produced by 46 U/L uricase action on urate. Unit activity is defined as 50% inhibition of maximal color development.
Within-run coefficients of variation (cv) were 2% for standards and samples, whereas between-run cv was 3.1% for standards and 7.3% for samples. Dilutional parallelism and linearity were demonstrated for 8-fold dilutions of samples over the range 0.1 to 1.1 U/mL. Recovery of added catalase was complete. Samples are stable to freezing and storage for 1 week at -80 degrees C. Activities (units/mL) ranged from 0.29 to 0.41 in human and canine plasma, and for erythrocytes from 48 to 70 in man, 17 to 19 in dogs, and 60 to 89 in rats. Rat liver activity (units/g wet weight) was age-dependent and ranged from 17 to 24 at 2 months, and from 19 to 37 at 6 months.
The first, fully automated assay for the measurement of catalatic activity of catalase in plasma, erythrocytes, and liver is demonstrated for multiple species. The assay is simple, precise, relatively inexpensive, and rapid.
开发过氧化氢酶抗氧化催化活性的全自动检测方法。
该检测基于临床化学自动分析仪的标准方法，使用Trinder试剂测定过氧化氢。过氧化氢酶与324 U/L辣根过氧化物酶（XII型）和Trinder试剂竞争46 U/L尿酸酶作用于尿酸产生的过氧化氢。单位活性定义为最大显色抑制50%。
标准品和样品的批内变异系数（cv）为2%，而标准品的批间cv为3.1%，样品的批间cv为7.3%。在0.1至1.1 U/mL范围内，样品8倍稀释显示出稀释平行性和线性。添加的过氧化氢酶回收完全。样品在-80℃冷冻和储存1周稳定。人及犬血浆中的活性（单位/mL）范围为0.29至0.41，人红细胞中的活性为48至70，犬红细胞中的活性为17至19，大鼠红细胞中的活性为60至89。大鼠肝脏活性（单位/克湿重）与年龄有关，2个月时为17至24，6个月时为19至37。
首次展示了一种用于多种物种血浆、红细胞和肝脏中过氧化氢酶催化活性测量的全自动检测方法。该检测方法简单、精确、相对便宜且快速。
