Plasma and dried blood spot lysosphingolipids for the diagnosis of different sphingolipidoses: a comparative study
Plasma and dried blood spot lysosphingolipids for the diagnosis of different sphingolipidoses: a comparative study
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DOI：
10.1515/cclm-2018-1301
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发表时间：
2019-12-01
影响因子：
6.8
通讯作者：
Burlina, Alberto B.
中科院分区：
文献类型：
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作者：
Polo, Giulia;Burlina, Alessandro P.;Burlina, Alberto B.
背景：溶血鞘脂是鞘脂的 N-脱酰基形式，已被确定为多种鞘脂沉积症的潜在生物标志物，例如戈谢病、法布里病、克拉布病和尼曼-皮克病以及 GM1 和 GM2 神经节苷脂病。迄今为止，已经开发出不同的方法来测量血浆中的各种溶血鞘脂 (LysoSL)。在这里，我们提出了一种新型液相色谱串联质谱 (LC-MS/MS) 测定法，用于同时定量干血斑 (DBS) 中的 LysoSL（HexSph、LysoGb3、LysoGM1、LysoGM2、LysoSM 和 LysoSM509）。该 LC-MS/MS 方法用于比较受影响患者和健康对照者 DBS 和血浆中 LysoSL 的水平。方法：从直径 3.2 mm 的 DBS 中提取溶血鞘脂，其中含有内部稳定同位素标准品的甲醇：乙腈：水（80:15:5，v/v）混合物。使用 C18 柱以水和乙腈的梯度进行色谱分离，两者均含有 0.1% 甲酸，总运行时间为 4 分钟。通过多反应监测 (MRM) 在正离子模式电喷雾电离 (ESI)-MS/MS 中检测到该化合物。结果：该方法在 DBS 上进行了验证，以证明特异性、线性、定量下限、准确度和精密度。参考范围是在儿童和成人人群中确定的。在戈谢病 (HexSph)、法布里病 (LysoGb3)、prosaposin 缺乏症（HexSph 和 LysoGb3）以及尼曼匹克病 A/B 和 C 型（LysoSM 和 LysoSM509）中发现了 LysoSL 水平升高。 LysoGb3 和 HexSph 的 DBS 与血浆水平之间的相关性非常好，但 LysoSM 和 LysoSM509 的相关性较差。结论：尽管血浆 LysoSL 测定仍然是金标准，但我们的 LC-MS/MS 方法可以快速可靠地定量 DBS 中的溶血鞘脂。该方法是诊断除-Niemann-Pick C 型以外的不同鞘脂沉积症的有用工具。
Background: Lysosphingolipids, the N-deacylated forms of sphingolipids, have been identified as potential biomarkers of several sphingolipidoses, such as Gaucher, Fabry, Krabbe and Niemann-Pick diseases and in GM1 and GM2 gangliosidoses. To date, different methods have been developed to measure various lysosphingolipids (LysoSLs) in plasma. Here, we present a novel liquid chromatography tandem mass spectrometry (LC-MS/MS) assay for a simultaneous quantification of LysoSLs (HexSph, LysoGb3, LysoGM1, LysoGM2, LysoSM and LysoSM509) in dried blood spot (DBS). This LC-MS/MS method was used to compare the levels of LysoSLs in DBS and plasma in both affected patients and healthy controls.Methods: Lysosphingolipids were extracted from a 3.2 mm diameter DBS with amixture of methanol:acetonitrile:water (80:15:5, v/v) containing internal stable isotope standards. Chromatographic separation was performed using a C18 column with a gradient of water and acetonitrile both with 0.1% formic acid in a total run time of 4 min. The-compounds were detected in the positive ion mode electrospray ionization (ESI)-MS/MS by multiple reaction monitoring (MRM).Results: The method was validated on DBS to demonstrate specificity, linearity, lowest limit of quantification, accuracy and precision. The reference ranges were determined in pediatric and adult populations. The elevated levels of LysoSLs were identified in Gaucher disease (HexSph), Fabry disease (LysoGb3), prosaposin deficiency (HexSph and LysoGb3) and Niemann-Pick disease types A/B and C (LysoSM and LysoSM509). The correlation in the levels between DBS and plasma was excellent for LysoGb3 and HexSph but poor for LysoSM and LysoSM509.Conclusions: Despite the fact that plasma LysoSLs determination remains the gold standard, our LC-MS/MS method allows a rapid and reliable quantification of lysosphingolipids in DBS. The method is a useful tool for the diagnosis of different sphingolipidoses except for -Niemann-Pick type C.