血红蛋白存在下胆碱酯酶活性的5,5′-二硫双-2-硝基苯甲酸分光光度测定法

5,5′-dithiobis-2-nitrobenzoic acid spectrophotometric method for determination of Acetylcholinesterase activity in presence of hemoglobin

5 ,5′二硫双 - 2 -硝基苯甲酸 (DTNB)分光光度测定法是检测乙酰胆碱酯酶酶活性的最灵敏的方法。但血液样品如兔网织红细胞裂解物样品中血红蛋白的存在会极大地干扰呈色反应 ,因此我们采用蛋白质变性剂三氯乙酸沉淀血红蛋白 ,再用NaOH将上清液校回中性的方法 ,对原DTNB法进行改良。改进后的DTNB法较原DTNB法大大降低了背景干扰 ,提高了检测灵敏度。

dithiobis-2-nitrobenzoic acid (DTNB) spectrophotome tric assay is the most sensitive method for determination of Acetylcholinesteras e (AChE) activity. However, the presence of hemoglobin (Hb) in blood samples, pr ofoundly influence the color development, thus we promoted the DTNB spectrophoto metric method by precipitated Hb using the denaturant trichoroacetic acid (TCA), and reneutralized the supernatant with NaOH. The modified DTNB method intensive ly decreased the background and increased the sensitivity.