Molybdenum Cofactor Deficiency: A New HPLC Method for Fast Quantification of S-Sulfocysteine in Urine and Serum
Molybdenum Cofactor Deficiency: A New HPLC Method for Fast Quantification of S-Sulfocysteine in Urine and Serum
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DOI：
10.1007/8904_2011_89
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Schwarz, Guenter
中科院分区：
文献类型：
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作者：
Belaidi, Abdel Ali;Arjune, Sita;Schwarz, Guenter
钼辅因子缺乏症（MoCD）是一种罕见的遗传性代谢性疾病，其特征是严重的进行性神经损伤，主要由亚硫酸盐氧化酶活性丧失引起。尿液中亚硫酸盐、硫代硫酸盐和 S-磺基半胱氨酸 (SSC) 水平升高是诊断 MoCD 和亚硫酸盐氧化酶缺乏症 (SOD) 的标志。最近，据报道，基于钼辅因子前体 cPMP 的替代疗法，首次成功治疗人类 MoCD A 型患者，导致 MoCD 生物标志物几乎完全正常化。了解未经治疗的患者疾病症状的快速进展、MoCD 的早期诊断以及监测 SSC 水平（亚硫酸盐毒性的关键标志物）日常变化的敏感方法对于治疗结果至关重要。在这里，我们描述了一种使用高效液相色谱 (HPLC) 分析人类尿液样本中 SSC 的快速、灵敏的方法。该分析基于邻苯二醛 (OPA) 的柱前衍生化和 C18 反相柱上的分离以及 UV 检测。该方法扩展到人血清分析，未发现内源氨基酸的干扰。最后，报告了来自对照个体以及 MoCD 患者的 45 份儿童尿液、75 份成人尿液和 24 份血清样本的 SSC 值。我们的方法代表了一种用于 MoCD 和 SOD 常规诊断的经济有效的技术，并且还可以用于每天监测这些亚硫酸盐毒性疾病的治疗效率。
Molybdenum cofactor deficiency (MoCD) is a rare inherited metabolic disorder characterized by severe and progressive neurological damage mainly caused by the loss of sulfite oxidase activity. Elevated urinary levels of sulfite, thiosulfate, and S-sulfocysteine (SSC) are hallmarks in the diagnosis of MoCD and sulfite oxidase deficiency (SOD). Recently, a first successful treatment of a human MoCD type A patient based on a substitution therapy with the molybdenum cofactor precursor cPMP has been reported, resulting in nearly complete normalization of MoCD biomarkers. Knowing the rapid progression of the disease symptoms in nontreated patients, an early diagnosis of MoCD as well as a sensitive method to monitor daily changes in SSC levels, a key marker of sulfite toxicity, is crucial for treatment outcome. Here, we describe a fast and sensitive method for the analysis of SSC in human urine samples using high performance liquid chromatography (HPLC). The analysis is based on precolumn derivatization with O-phthaldialdehyde (OPA) and separation on a C18 reverse phase column coupled to UV detection. The method was extended to human serum analysis and no interference with endogenous amino acids was found. Finally, SSC values from 45 pediatric urine, 75 adult urine, and 24 serum samples from control individuals as well as MoCD patients are reported. Our method represents a cost-effective technique for routine diagnosis of MoCD and SOD, and can be used also to monitor treatment efficiency in those sulfite toxicity disorders on a daily basis.