McComb R B, Bond L W, Burnett R W, Keech R C, Bowers G N
Clin Chem. 1976 Feb;22(2):141-50.
The molar absorptivity of NADH at 340 nm has been determined by an indirect procedure in which high-purity glucose is phosphorylated by ATP in the presence of hexokinase, coupled to oxidation of the glucose-6-phosphate by NAD+ in the presence of glucose-6-phosphate dehydrogenase. The average value from 85 independent determinations is 6317 liter mol-1 cm-1 at 25 degrees C and pH 7.8. The overall uncertainty is -4.0 to +5.5 ppt (6292 to 6352 liter mol-1 cm-1), based on a standard error of the mean of 0.48 ppt and an estimate of systematic error of -2.6 to +4.1 ppt. Effects of pH, buffer, and temperature on the molar absorptivity are also reported.
NADH在340nm处的摩尔吸光系数已通过间接方法测定。在该方法中，高纯度葡萄糖在己糖激酶存在下被ATP磷酸化，并在葡萄糖-6-磷酸脱氢酶存在下与NAD⁺将葡萄糖-6-磷酸氧化相偶联。在25℃和pH 7.8条件下，85次独立测定的平均值为6317升·摩尔⁻¹·厘米⁻¹。基于平均标准误差0.48ppt和系统误差估计值-2.6至+4.1ppt，总体不确定度为-4.0至+5.5ppt（6292至6352升·摩尔⁻¹·厘米⁻¹）。还报告了pH、缓冲液和温度对摩尔吸光系数的影响。
