Qi X Y, Keyhani N O, Lee Y C
Biology Department, Johns Hopkins University, Baltimore, Maryland 21218.
Anal Biochem. 1988 Nov 15;175(1):139-44. doi: 10.1016/0003-2697(88)90371-5.
A method has been developed to measure hydrazine, hydrazides, and their mixtures using a modification of the trinitrobenzenesulfonic acid method [T. Okuyama and K. Satake (1960) J. Biochem. (Tokyo) 47, 654-660]. After incubation of the sample containing hydrazine and hydrazide with trinitrobenzenesulfonate at pH 8.5 at room temperature for 40 min, the reaction mixture was diluted with a Na2CO3-NaHCO3 buffer (0.1 M, pH 10.8) rather than with 0.5 M HCl. Different chromogens were produced from the reaction of hydrazine (lambda max = 570 nm) and hydrazides (lambda max = 385 and 500 nm) with trinitrobenzenesulfonic acid. The method allowed simultaneous determination of hydrazine (5 to 60 nmol) with hydrazide (10 to 120 nmol) in a mixture with a standard deviation of less than 5%. The presence of amino compounds (except for amino sugars) did not interfere with the measurement of hydrazine or hydrazides. Interference by amino sugars in the determination of hydrazine or hydrazides was eliminated by pretreatment of the sample with NaBH4 to reduce the amino sugars to 2-amino-2-deoxy-hexitols.
已开发出一种方法，通过对三硝基苯磺酸法进行改进来测定肼、酰肼及其混合物[T. Okuyama和K. Satake（1960年），《生物化学杂志》（东京）47, 654 - 660]。将含有肼和酰肼的样品与三硝基苯磺酸盐在室温下于pH 8.5孵育40分钟后，反应混合物用Na2CO3 - NaHCO3缓冲液（0.1 M，pH 10.8）稀释，而不是用0.5 M HCl稀释。肼（最大吸收波长 = 570 nm）和酰肼（最大吸收波长 = 385和500 nm）与三硝基苯磺酸反应产生不同的显色剂。该方法可同时测定混合物中的肼（5至60 nmol）和酰肼（10至120 nmol），标准偏差小于5%。氨基化合物（氨基糖除外）的存在不干扰肼或酰肼的测定。通过用NaBH4对样品进行预处理将氨基糖还原为2 - 氨基 - 2 - 脱氧 - 己糖醇，可消除氨基糖对肼或酰肼测定的干扰。
