Lazzarino Giuseppe, Amorini Angela Maria, Di Pietro Valentina, Tavazzi Barbara
Division of Biochemistry and Molecular Biology, Department of Chemical Sciences, University of Catania, Catania, Italy.
Methods Mol Biol. 2011;708:99-117. doi: 10.1007/978-1-61737-985-7_5.
The determination of purines and pyrimidines in biofluids is useful for the clinical-biochemical characterization of acute and chronic pathological states that induce transient or permanent alterations of metabolism. In particular, the diagnosis of several inborn errors of metabolism (IEMs) is accomplished by the analysis of circulating and excreted purines and pyrimidines. It is certainly advantageous to simultaneously determine the full purine and pyrimidine profile, as well as to quantify other compounds of relevance (e.g., organic acids, amino acids, sugars) in various metabolic hereditary diseases, in order to screen for a large number of IEMs using a reliable and sensitive analytical method characterized by mild to moderate costs. Toward this end, we have developed an ion-pairing HPLC method with diode array detection for the synchronous separation of several purines and pyrimidines. This method also allows the quantification of additional compounds such as N-acetylated amino acids and dicarboxylic acids, the concentrations of which are profoundly altered in different IEMs. The application of the method in the analysis of biological samples from patients with suspected purine and pyrimidine disorders is presented to illustrate its applicability for the clinical-biochemical diagnosis of IEM.
生物流体中嘌呤和嘧啶的测定，对于诱导代谢出现短暂或永久性改变的急慢性病理状态的临床生化特征分析很有用。特别是，几种先天性代谢缺陷（IEMs）的诊断是通过分析循环和排泄的嘌呤和嘧啶来完成的。在各种代谢遗传性疾病中，同时测定完整的嘌呤和嘧啶谱以及定量其他相关化合物（如有机酸、氨基酸、糖类），以便使用一种可靠且灵敏、成本适中的分析方法筛查大量IEMs，这无疑是有利的。为此，我们开发了一种采用二极管阵列检测的离子对高效液相色谱法，用于同步分离多种嘌呤和嘧啶。该方法还能够对其他化合物进行定量，如N-乙酰化氨基酸和二羧酸，它们的浓度在不同的IEMs中会发生显著变化。本文介绍了该方法在疑似嘌呤和嘧啶紊乱患者生物样品分析中的应用，以说明其在IEM临床生化诊断中的适用性。
