Development of a new chromogenic method for putrescine quantification using coupling reactions involving putrescine oxidase
Development of a new chromogenic method for putrescine quantification using coupling reactions involving putrescine oxidase
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DOI：
10.1016/j.ab.2020.113607
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发表时间：
2020-03-15
影响因子：
2.9
通讯作者：
Kurihara, Shin
中科院分区：
文献类型：
--
作者：
Sugiyama, Yuta;Ohta, Hirokazu;Kurihara, Shin
包括腐胺在内的多胺的定量通常使用高效液相色谱或气相色谱进行。然而，由于样品的衍生化和分析试剂的制备，这些方法非常耗时。在这项研究中，我们建立了一种简单、高通量的腐胺定量方法，在96孔板上使用来自红球红球菌NCIMB 11540的腐胺氧化酶、过氧化物酶、4-氨基安替比林和N-乙基-N-(3-磺丙基)-3-甲基苯胺钠盐。该方法(命名为PuO-POD-4AA-TOPS法)适用于细菌培养上清。此外，用改进的方法测定的腐胺浓度与用高效液相色谱法测定的浓度大致一致。
Quantification of polyamines, including putrescine, is generally performed using high-performance liquid chromatography (HPLC) or gas chromatography. However, these methods are time-consuming because of sample derivatization and analytical reagent preparation. In this study, we developed a simple and high-throughput putrescine quantification method on a 96-well microtiter plate using putrescine oxidase from Rhodococcus erythropolis NCIMB 11540, peroxidase, 4-aminoantipyrine, and N-ethyl-N-(3 sulfopropyl)-3-methylaniline sodium salt. The developed method (named as PuO-POD-4AA-TOPS method) was applicable to bacterial culture supernatants. Furthermore, putrescine concentrations determined by the developed method roughly corresponded to the concentrations determined by HPLC.