维多利亚蓝4R褪色分光光度法测定辅酶Ⅰ  

Fading Spectrophotometric Determination of Two Coenzyme I with Victoria Blue 4R

作　　者：刘林丰[1] 刘忠芳[1] 刘绍璞[1] 段慧[1] 

机构地区：[1]西南大学化学化工学院、发光与实时分析教育部重点实验室,重庆400715

出　　处：《西南大学学报（自然科学版）》2009年第1期60-63,共4页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(20475045)

摘　　要：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 interaction between basic diphenylmethane dyes, i.e. Night Blue （NB）, Victoria Blue B （VBB） and Victoria Blue 4R （VB4R） with two types of Coenzyme Ⅰ （NAD＋ and NADH） was studied by a spectrophotometry. The results showed that the absorption spectra changed only when CoI reacted with Victoria Blue 4R, resulting in an obvious decrease in absorptivity for VB4R at the maximum wavelength 611 nm. The fading degree and the concentration of CoI were followed by the law of Lambert-Beer＇s in the quantitative determination ranges of 1.07 - 4.00 μg/mL （NAD^＋ ） and 0.45 - 4.00μg/mL （NADH）. Their maximum molar absorption coefficients （ε） were 3.92×10^4 L/（mol·cm） for NAD＋ and 6.62×10^5 L/（mol·cm） for NADH. In this work, the characteristics of the absorption spectra, the optimum reaction conditions and the effects of coexisting substances were investigated, and the results indicated that the proposed method has a good selectivity and is recommended for application in the determination of NADH in synthetic water samples.

关 键 词：辅酶Ⅰ 二苯萘基甲烷类染料 分光光度法 

分 类 号：O657.3[理学—分析化学]