RESORUFIN BUTYRATE + INDOXYL ACETATE AS FLUOROGENIC SUBSTRATES FOR CHOLINESTERASE
RESORUFIN BUTYRATE + INDOXYL ACETATE AS FLUOROGENIC SUBSTRATES FOR CHOLINESTERASE
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DOI：
10.1021/ac60220a031
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发表时间：
1965-01-01
影响因子：
7.4
通讯作者：
KRAMER, DN
中科院分区：
文献类型：
--
作者：
GUILBAULT, GG;KRAMER, DN
本文报道了两种新的测定胆碱酯酶的荧光底物试卤灵丁酸酯和吲哚酚乙酸酯。两者都是非荧光化合物，它们被胆碱酯酶水解成高荧光物质。测量由于试卤灵（[λ]ex = 540-570 m[mu]，[λ]em = 580 m[mu]）和吲哚酚（[λ]ex = 395 m[mu]，[λ]em = 470 m[mu]）的产生而引起的溶液荧光的变化速率，[Δ]F/[Δ]t，并将其与酶活性相关联。底物溶液的制备相对简单，完全分析仅需2至5分钟。采用所述方法，可以测定0.00030至0.060单位/毫升的马血清胆碱酯酶，相对标准偏差约为[±] 1%。除了胆碱酯酶，酰化酶，酸性磷酸酶，胰凝乳蛋白酶在不同程度上水解底物。
Two new fluorogenic substrates are described for the determination of cholinesterase resorufin butyrate and indoxyl acetate. Both are nonfluorescent compounds which are hydrolyzed by cholinesterase to highly fluorescent materials. The rate of change in the fluorescence of the solution due to production of resorufin ([lambda]ex = 540-570 m[mu], [lambda]em = 580 m[mu]) and indoxyl ([lambda]ex = 395 m[mu], [lambda]em = 470 m[mu]), [DELTA]F/[DELTA]t, is measured and correlated with enzyme activity. The preparation of the substrate solutions is relatively simple, and complete analysis requires only 2 to 5 minutes. Employing the methods described, 0.00030 to 0.060 unit per milliliter of horse serum cholinesterase may be determined with a relative standard deviation of about [plus or minus]1%. In addition to cholinesterase, acylase, acid phosphatase, and chymotrypsin hydrolyzed the substrates in varying degrees.