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使用气相色谱/质谱联用技术同时测量尿中多元醇。

Simultaneous measurement of urinary polyols using gas chromatography/mass spectrometry.

作者信息

Lee Jeongae, Chung Bong Chul

机构信息

Bioanalysis and Biotransformation Research Center, Korea Institute of Science and Technology, Haweolgok-Dong 39-1, Sungbuk-Ku, Seoul 136-791, Republic of Korea.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Feb 2;831(1-2):126-31. doi: 10.1016/j.jchromb.2005.11.043. Epub 2005 Dec 13.

Abstract

In the present study, we simultaneously measured several polyols, such as adonitol, arabitol, dulcitol, glucose, myo-inositol, mannitol, sorbitol, and xylitol, in urine by gas chromatography/mass spectrometry-positive chemical ionization. We also examined possible relationship between the levels of these metabolites and age in normal individuals. In order to proceed to its quantification by GC/MS, 200 microL of a urine sample were diluted with 3 ml of distilled water, lyophilized, acetylated, and then analyzed them. Using this method, we were able to quantify as little as 0.5-1.0 ng/microL, and we made the calibration curves to be linear from 0.25 to 250 ng/microL (r(2)>0.991). Analytical recoveries were over 89.4%, and the inter-day and intra-day variability for accuracy and reproducibility was less than 20%. In the normal urine sample, the levels of polyols were gender-differentiated and age-related. This simple GC/MS method is sensitive and allows the measurement of wide ranges of polyols using small amounts of urine. We conclude that the quantitation of urinary polyols using GC/MS appears to be a clinically useful method for assessing polyol-pathway activity.

摘要

在本研究中,我们通过气相色谱/质谱-正化学电离法同时测定了尿液中的几种多元醇,如阿东糖醇、阿拉伯糖醇、卫矛醇、葡萄糖、肌醇、甘露醇、山梨醇和木糖醇。我们还研究了这些代谢物水平与正常个体年龄之间的可能关系。为了通过气相色谱/质谱进行定量分析,将200微升尿液样本用3毫升蒸馏水稀释,冻干,乙酰化,然后进行分析。使用这种方法,我们能够低至0.5-1.0纳克/微升进行定量,并且我们绘制了校准曲线,使其在0.25至250纳克/微升范围内呈线性(r²>0.991)。分析回收率超过89.4%,日间和日内准确度及重现性的变异性小于20%。在正常尿液样本中,多元醇水平存在性别差异且与年龄相关。这种简单的气相色谱/质谱方法灵敏,能够使用少量尿液测量多种多元醇。我们得出结论,使用气相色谱/质谱对尿中多元醇进行定量似乎是评估多元醇途径活性的一种临床有用方法。

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