Detection of novel visible-light region absorbance peaks in the urine after alkalization in patients with alkaptonuria.
Detection of novel visible-light region absorbance peaks in the urine after alkalization in patients with alkaptonuria.
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碱性藻尿症患者的浸泡后，尿液中新型明显区域吸光度峰的检测。
DOI：
10.1371/journal.pone.0086606
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Shimosawa T
中科院分区：
文献类型：
--
作者：
Tokuhara Y;Shukuya K;Tanaka M;Mouri M;Ohkawa R;Fujishiro M;Takahashi T;Okubo S;Yokota H;Kurano M;Ikeda H;Yamaguchi S;Inagaki S;Ishige-Wada M;Usui H;Yatomi Y;Shimosawa T
尿黑酸尿症由尿黑酸1，2-双加氧酶缺乏引起，导致尿黑酸（2，5-二羟基苯乙酸，HGA）在尿中积累。当尿液在室温下静置数小时至数天后发生颜色变化时，怀疑是尿黑酸尿症;尿黑酸氧化为苯醌乙酸是这种颜色变化的基础，加入碱会加速这种颜色变化。为了开发一种简便的黑酸尿症筛查试验，我们将碱加入到从黑酸尿症患者获得的尿液样品中，并测量可见光区域的吸收光谱。我们评价了尿黑酸尿症患者（n = 2）的尿液样品的吸收光谱的特征，并将其与健康志愿者（n = 5）和苯丙酮尿症患者（n = 3）的尿液样品以及碱化后的合成尿黑酸溶液的吸收光谱进行了比较。 通过加入NaOH、KOH或NH 4 OH对尿样和HGA溶液进行碱化。将样品溶液在室温下孵育1 min，然后测量吸收光谱。碱性尿中加入碱在406 nm和430 nm处产生特征吸收峰，碱化后的HGA溶液也得到相同的结果。在406 nm和430 nm处的吸光度值均以时间依赖性方式增加。此外，与弱碱性样品（添加NH 4 OH）相比，强碱性样品（添加NaOH-KOH）在这些峰处的吸光度值更大。此外，向样品中加入抗坏血酸后，峰消失。碱化后的尿黑酸和HGA溶液在406 nm和430 nm处均出现两个特征峰。这种新的快速简便的方法可能为发展一种简便的诊断黑尿症的方法铺平道路。
Alkaptonuria, caused by a deficiency of homogentisate 1,2-dioxygenase, results in the accumulation of homogentisic acid (2,5-dihydroxyphenylacetic acid, HGA) in the urine. Alkaptonuria is suspected when the urine changes color after it is left to stand at room temperature for several hours to days; oxidation of homogentisic acid to benzoquinone acetic acid underlies this color change, which is accelerated by the addition of alkali. In an attempt to develop a facile screening test for alkaptonuria, we added alkali to urine samples obtained from patients with alkaptonuria and measured the absorbance spectra in the visible light region. We evaluated the characteristics of the absorption spectra of urine samples obtained from patients with alkaptonuria (n = 2) and compared them with those of urine specimens obtained from healthy volunteers (n = 5) and patients with phenylketonuria (n = 3), and also of synthetic homogentisic acid solution after alkalization. Alkalization of the urine samples and HGA solution was carried out by the addition of NaOH, KOH or NH4OH. The sample solutions were incubated at room temperature for 1 min, followed by measurement of the absorption spectra. Addition of alkali to alkaptonuric urine yielded characteristic absorption peaks at 406 nm and 430 nm; an identical result was obtained from HGA solution after alkalization. The absorbance values at both 406 nm and 430 nm increased in a time-dependent manner. In addition, the absorbance values at these peaks were greater in strongly alkaline samples (NaOH- KOH-added) as compared with those in weakly alkaline samples (NH4OH-added). In addition, the peaks disappeared following the addition of ascorbic acid to the samples. We found two characteristic peaks at 406 nm and 430 nm in both alkaptonuric urine and HGA solution after alkalization. This new quick and easy method may pave the way for the development of an easy method for the diagnosis of alkaptonuria.
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影响因子：
56.9
作者：
SEALY, RC;HYDE, JS;PROTA, G
通讯作者：
PROTA, G
影响因子：
158.5
作者：
Phornphutkul, C;Introne, WJ;Gahl, WA
通讯作者：
Gahl, WA
影响因子：
7.4
作者：
MARTIN, JP;BATKOFF, B
通讯作者：
BATKOFF, B
影响因子：
5.9
作者：
OBRIEN, WM;LADU, BN;BUNIM, JJ
通讯作者：
BUNIM, JJ
影响因子：
4.1
作者：
CONSDEN, R;FORBES, HAW;STANIER, WM
通讯作者：
STANIER, WM