用于检测来自硫酸角质素的相关二糖作为黏多糖贮积症IVA型综合征生物标志物的液相色谱-串联质谱法检测的验证。
Validation of an LC-MS/MS assay for detecting relevant disaccharides from keratan sulfate as a biomarker for Morquio A syndrome.
作者信息
Martell Lisa Argento, Cunico Robert L, Ohh Jayoung, Fulkerson Wendy, Furneaux Richard, Foehr Erik D
机构信息
BioMarin Pharmaceutical Inc., Novato, CA 94949, USA.
出版信息
Bioanalysis. 2011 Aug;3(16):1855-66. doi: 10.4155/bio.11.172.
BACKGROUND
Mucopolysaccharidosis IVA (MPS IVA, Morquio A syndrome) is an inherited lysosomal storage disease caused by deficiency of N-acetylgalactosamine-6-sulfatase (GALNS), an enzyme required for stepwise degradation of keratan sulfate (KS). We have developed a selective, sensitive, accurate and precise LC-MS/MS assay for the KS-derived disaccharides Galβ1-4GlcNAc(6S) and Gal(6S)β1-4GlcNAc(6S) in human urine and plasma using keratanase II digestion.
RESULTS
Mean accuracy was 96-106% in urine and 97-108% in plasma. Precision was high, with relative standard deviations of 1-2% (intra-day) and 2-5% (inter-day) in urine and 1-2% (intra-day) and 4-7% (inter-day) in plasma. The lower limit of quantitation was 0.026 µg/ml (plasma) and 0.104 µg/ml (urine), with a quantitation range of 0.026-5 µg/ml (plasma) and 0.104-20 µg/ml (urine).
CONCLUSION
Clinical sample analysis in 168 MPS IVA patients and 225 healthy controls demonstrates the clinical utility of this method.
背景
黏多糖贮积症IVA型（MPS IVA，莫尔基奥A综合征）是一种遗传性溶酶体贮积病，由N - 乙酰半乳糖胺 - 6 - 硫酸酯酶（GALNS）缺乏引起，GALNS是逐步降解硫酸角质素（KS）所需的一种酶。我们利用角质酶II消化法开发了一种用于检测人尿液和血浆中KS衍生二糖Galβ1 - 4GlcNAc(6S)和Gal(6S)β1 - 4GlcNAc(6S)的选择性、灵敏、准确且精密的液相色谱 - 串联质谱（LC - MS/MS）检测方法。
结果
尿液中的平均准确度为96 - 106%，血浆中的平均准确度为97 - 108%。精密度较高，尿液中日内相对标准偏差为1 - 2%，日间相对标准偏差为2 - 5%；血浆中日内相对标准偏差为1 - 2%，日间相对标准偏差为4 - 7%。定量下限为0.026 µg/ml（血浆）和0.104 µg/ml（尿液），定量范围为0.026 - 5 µg/ml（血浆）和0.104 - 20 µg/ml（尿液）。
结论
对168例MPS IVA患者和225例健康对照的临床样本分析证明了该方法的临床实用性。
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