Efficient analysis of urinary glycosaminoglycans by LC-MS/MS in mucopolysaccharidoses type I, II and VI
Efficient analysis of urinary glycosaminoglycans by LC-MS/MS in mucopolysaccharidoses type I, II and VI
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DOI：
10.1016/j.ymgme.2010.09.003
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发表时间：
2011-01-01
影响因子：
3.8
通讯作者：
Millington, David S.
中科院分区：
文献类型：
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作者：
Auray-Blais, Christiane;Bherer, Patrick;Millington, David S.
粘多糖贮积症（MPS）是由特异性溶酶体酶缺乏引起的复杂的储存障碍，导致糖胺聚糖（GAGS）在尿液、血浆以及各种组织中积聚。我们设计并验证了一种简单，但准确和精确的串联质谱方法，结合高效液相色谱法（LC-MS/MS），用于定量尿液中的GAG。该方法适用于MPS I、II和VI患者的研究，通过定量尿液中的硫酸皮肤素（DS）和硫酸乙酰肝素（HS）。我们分析了28例MPS患者（年龄1至42岁）和55例对照受试者（41天至18岁）的尿液样本。所有年龄健康对照的尿液中DS和HS水平均低于定量限。6例接受治疗的MPS I患者的DS（0.7-45 vs 9.3-177 mg/mmol肌酐）和HS（0-123 mg/mmol肌酐vs 38-418 mg/mmol肌酐）尿液中DS和HS水平低于6例未接受治疗的MPS I患者; 9例MPS II患者和7例MPS VI患者获得了相似的结果。对少至250 μ L的尿液进行分析。甲醇分解每个样品需要75分钟;每次LC-MS/MS进样的总分析运行时间为8分钟。结果表明，该方法适用于各种各样的情况下，需要高准确度和精密度，包括现有和新兴的治疗的有效性评价。(C)2010年爱思唯尔公司All rights reserved.
Mucopolysaccharidoses (MPSs) are complex storage disorders caused by specific lysosomal enzyme deficiencies, resulting in the accumulation of glycosaminoglycans (GAGS) in urine, plasma, as well as in various tissues. We devised and validated a straightforward, but accurate and precise tandem mass spectrometry methodology coupled to high performance liquid chromatography (LC-MS/MS) for the quantification of GAGs in urine. The method is applicable to the investigation of patients with MPS I, II, and VI, by quantifying dermatan sulfate (DS) and heparan sulfate (HS) in urine. We analyzed urine samples from 28 MPS patients, aged 1 to 42 years, and 55 control subjects (41 days to 18 years old). Levels of DS and HS in urine from healthy controls of all ages were below the limit of quantification. The levels of DS and HS in urine from 6 treated patients with MPS I were lower than in 6 untreated patients in DS (0.7-45 vs 9.3-177 mg/mmol creat) and HS (0-123 mg/mmol creatinine vs 38-418 mg/mmol creatinine); similar results were obtained for 9 patients with MPS II and 7 patients with MPS VI. Analyses were performed on as little as 250 mu L of urine. Methanolysis took 75 min per sample; the total analysis run time for each LC-MS/MS injection was 8 min. Results indicate that the method is applicable to a wide variety of situations in which high accuracy and precision are required, including the evaluation of the effectiveness of existing and emerging treatments. (C) 2010 Elsevier Inc. All rights reserved.