Matsubara C, Yokoi Y, Nakamichi N, Takamura K
Tokyo College of Pharmacy, Japan.
Yakugaku Zasshi. 1994 Jan;114(1):48-53. doi: 10.1248/yakushi1947.114.1_48.
Aqueous solution of oxo[5,10,15,20-tetra(4-pyridyl)porphyrinato]titanium (IV) complex, a Ti-TPyP reagent, was found to be very useful for the spectrophotometric determination of hydrogen peroxide. The reagent (lambda max 432 nm) reacts with hydrogen peroxide to form a monoperoxocomplex, resulting in a significant decrease of the absorbance at 432 nm. The decrease (delta A) in absorbance was proportional to the concentration of hydrogen peroxide. The Ti-TPyP reagent was successfully applied to the assay of uric acid in the serum, using uricase to produce hydrogen peroxide through enzymatic oxidation. Using only 5 microliters serum, a linear relationship was obtained between delta A and uric acid concentration in the serum ranging from 5 x 10(-6) to 1 x 10(-3) M. The apparent molar delta A of uric acid was 2.2 x 10(5) M-1 cm-1. The relative standard deviation of repeated runs (n = 8) was 2.8% at 3.77 x 10(-4) M uric acid. The analytical recovery of uric acid (5 x 10(-4) M) added to the serum was 96.8 to 105.0%. No pre-concentration and deproteinization were required to determine uric acid in the serum by the present method because of the high sensitivity and selectivity of the Ti-TPyP reagent for hydrogen peroxide.
发现氧代[5,10,15,20-四(4-吡啶基)卟啉合]钛(IV)配合物水溶液(一种Ti-TPyP试剂)对分光光度法测定过氧化氢非常有用。该试剂(最大吸收波长432 nm)与过氧化氢反应形成单过氧配合物,导致432 nm处吸光度显著降低。吸光度的降低(ΔA)与过氧化氢浓度成正比。Ti-TPyP试剂通过尿酸酶催化氧化产生过氧化氢,成功应用于血清中尿酸的测定。仅使用5微升血清,在血清尿酸浓度为5×10⁻⁶至1×10⁻³ M范围内,ΔA与尿酸浓度之间获得了线性关系。尿酸的表观摩尔ΔA为2.2×10⁵ M⁻¹ cm⁻¹。在尿酸浓度为3.77×10⁻⁴ M时,重复测定(n = 8)的相对标准偏差为2.8%。添加到血清中的尿酸(5×10⁻⁴ M)的分析回收率为96.8%至105.0%。由于Ti-TPyP试剂对过氧化氢具有高灵敏度和选择性,因此采用本方法测定血清中的尿酸无需预浓缩和脱蛋白。