Method of measuring phenol and alpha-naphthol, and method of measuring the activity of enzymes
申请人:SANWA KAGAKU KENKYUSHO CO., LTD.
公开号:EP0243125A2
公开(公告)日:1987-10-28
To measure the amount of phenol or α-naphthol in an aqueous sample, the sample is (a) mixed in aqueous solution with a diamine (preferably an aniline e.g. having a para- amino group, e.g. p-diethylaminoaniline), in the presence of an oxidant (e.g. potassium ferricyanide, chloramine T, Na metaperiodate or H202 and a peroxidase) to form a quinone dye having a maximum absorption near 670 nm, and (b) measuring the depth of color (absorbency) formed compared to that of a control. The concentration of phenol or naphthol can be calculated from the absorbancy of a standard solution.
The test sample can be one obtained from the action of an enzyme (e.g. an alkaline or acidic phosphatase, kallikrein or antithrombin III on a substrate (e.g. disodium phenyl phosphate) from which phenol or α-naphthol is liberated enzymatically; the enzyme activity can thus be determined. A reagent test paper can be dipped in a liquid sample (e.g. of urine) and developed with a drop of oxidant solution.
The method is rapid, sensitive and highly reproducible.
为了测量水溶液样品中的苯酚或α-萘酚含量,样品(a)在氧化剂(如铁氰化钾、氯胺 T、偏碘酸纳或 H202 和过氧化物酶)的存在下,与二胺(最好是苯胺,如具有对氨基的苯胺,如对二乙氨基苯胺)在水溶液中混合,形成具有 670 纳米附近最大吸收的醌染料,(b)测量酚或α-萘酚的含量。例如铁氰化钾、氯胺 T、Na metaperiodate 或 H202 和过氧化物酶)的存在下,形成最大吸收波长接近 670 纳米的醌染料,以及 (b) 与对照组相比,测量形成的颜色深度(吸光度)。苯酚或萘酚的浓度可以通过标准溶液的吸光度计算出来。
测试样品可以是一种酶(如碱性或酸性磷酸酶、卡利克林或抗凝血酶 III)作用于底物(如苯基磷酸二钠)后得到的样品,苯酚或 α-萘酚就是从这种底物中酶解出来的;因此可以测定酶的活性。将试纸浸入液体样本(如尿液)中,滴入一滴氧化剂溶液即可显影。
该方法快速、灵敏、重复性高。