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用分光光度法研究了夜蓝(NB)、维多利亚蓝B(VBB)和维多利亚蓝4R(VB4R)3种碱性二苯基萘基染料与氧化型辅酶Ⅰ(NAD+)和还原型辅酶Ⅰ(NADH)的相互作用,结果表明仅有维多利亚蓝4R与两种辅酶Ⅰ反应引起吸收光谱的变化,并在611nm处产生明显的褪色反应.褪色反应分别在1.07～4.00μg/mL(NAD+),0.45～4.00μg/mL(NADH)浓度范围内遵循比尔定律,其摩尔吸光系数(ε)分别为3.92×104L/(mol·cm)(NAD+)和6.62×104L/(mol·cm)(NADH).研究了褪色反应的吸收光谱特征、适宜的反应条件、影响因素,考察了共存物质的影响,表明方法有较好的选择性.方法可用于合成水样中还原型辅酶Ⅰ的测定.
The effects of three basic diphenylnaphthyl dyes, such as NB, VBB and VB4R, with oxidized coenzyme Ⅰ (NAD +) and reduced coenzyme Ⅰ (NADH) were investigated by spectrophotometry. ) Results showed that only the reaction between Victoria Blue 4R and the two coenzyme Ⅰ caused the change of absorption spectrum and the obvious discoloration reaction at 611nm.The fading reaction was in the range of 1.07 ~ 4.00μg / mL (NAD +), 0.45 ~ The molar absorptivity (ε) of 4.00 μg / mL (NADH) was 3.92 × 104 L / (mol · cm) (NAD +) and 6.62 × 104 L / (mol · cm) The absorption spectrum of fading reaction, the suitable reaction conditions and influential factors were investigated, and the influence of coexisting substances was also investigated, indicating that the method has good selectivity. The method can be used for the determination of reduced coenzyme Ⅰ in synthetic water samples.