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基于添加五水合次氯酸钠氧化的分光光度法测定色氨酸。

A spectrophotometric method for the determination of tryptophan following oxidation by the addition of sodium hypochlorite pentahydrate.

机构信息

Department of Medical Technology, Ehime Prefectural University of Health Sciences, Ehime, Japan.

Department of Medical Technology, Kagawa Prefectural University of Health Sciences, Kagawa, Japan.

出版信息

PLoS One. 2023 Jan 26;18(1):e0279547. doi: 10.1371/journal.pone.0279547. eCollection 2023.

Abstract

Tryptophan (Trp) is an essential amino acid that functions in various biological processes and human daily health. As the significant functions of Trp become more apparent, its measurement is becoming increasingly important in various situations. Herein, we improved the Trp color reaction based on the Hopkins-Cole reaction and established a simple colorimetric method for Trp determination using several different reagents, including sodium hypochlorite pentahydrate and monosodium glutamate. The detection method can be performed using safe materials, rather than conventional toxic substances, and induces a crimson color change with an absorption peak at 525 nm, enabling the quantification of Trp by simple spectrophotometry in just 10 min. This assay exhibited a linear detection range from 10 to 100 mg/L (R2 = 0.9996). The average recoveries in the spiked cerebrospinal fluid ranged from 90.5% to 104.3%, with a relative standard deviation of 0.27% (n = 3, 29.40 mg/L Trp) to 1.19% (n = 3, 72.90 mg/L Trp). This novel spectrophotometric method may enable many researchers and laboratory technicians to detect Trp in various sample solutions without expensive analytical instruments or complicated operations.

摘要

色氨酸(Trp)是一种必需氨基酸,在各种生物过程和人类日常健康中发挥作用。随着 Trp 的重要功能变得更加明显,在各种情况下对其进行测量变得越来越重要。在此,我们基于 Hopkins-Cole 反应改进了 Trp 的显色反应,并使用几种不同的试剂(包括五水合次氯酸钠和谷氨酸单钠)建立了一种简单的比色法来测定 Trp。该检测方法可以使用安全的材料,而不是传统的有毒物质,并且会引起深红色的颜色变化,在 525nm 处有吸收峰,通过简单的分光光度法在 10 分钟内即可定量 Trp。该测定法的线性检测范围为 10 至 100mg/L(R2=0.9996)。在添加的脑脊液中的平均回收率在 90.5%至 104.3%之间,相对标准偏差为 0.27%(n=3,29.40mg/L Trp)至 1.19%(n=3,72.90mg/L Trp)。这种新颖的分光光度法可以使许多研究人员和实验室技术人员无需昂贵的分析仪器或复杂的操作即可检测各种样品溶液中的 Trp。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ef3/9879471/0a2c01f12260/pone.0279547.g001.jpg

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