Evaluation of a Liquid Chromatography–Tandem Mass Spectrometry Method for the Analysis of Glucosylceramide and Galactosylceramide Isoforms in Cerebrospinal Fluid of Parkinson’s Disease Patients,Analytical Chemistry

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Evaluation of a Liquid Chromatography–Tandem Mass Spectrometry Method for the Analysis of Glucosylceramide and Galactosylceramide Isoforms in Cerebrospinal Fluid of Parkinson’s Disease Patients
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-07-24 , DOI: 10.1021/acs.analchem.4c02654
Laura Castillo-Ribelles _{1,

2,

3} , Jose Antonio Arranz-Amo _{1,

2} , Jorge Hernández-Vara _{4,

5} , Daniela Samaniego-Toro ₅ , Silvia Enriquez-Calzada ₄ , Sara Lucas-Del Pozo _{4,

5,

6} , Maria Camprodon-Gomez _{4,

7} , Ariadna Laguna _{3,

4} , Mercedes Arrúe Gonzalo _{3,

4} , Roser Ferrer _{1,

2,

3} , Marta Martinez-Vicente _{3,

4} , Clara Carnicer-Caceres _{1,

2}

Affiliation

Mutations in GBA1, encoding glucocerebrosidase beta 1 (GCase), are the most common genetic risk factor for Parkinson’s disease (PD). GCase dysfunction leads to an accumulation of glucosylceramide (GluCer) substrates in different organs and fluids. Despite the challenges in quantifying GluCer isoforms in biological samples, their potential clinical interest as PD biomarkers justifies the development of robust assays. An extensively evaluated high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) method for quantifying 14 GluCer and galactosylceramide (GalCer) isoforms in human cerebrospinal fluid (CSF) samples is presented. Sample pretreatment, HPLC, and MS/MS parameters were optimized. Evaluation was performed according to the recommendations of the Clinical and Laboratory Standards Institute and European Medicines Agency guidelines. Four 7-point calibration curves were generated, with a linearity interval from 2.5 to 200 nM (R² ≥ 0.995). The limit of quantification was set at 5 nM. Between-run precision and accuracy were up to 12.5 and 9%, respectively. After method validation, we measured the levels of GluCer and GalCer isoforms in CSF human samples, including 6 healthy controls (HC), 22 idiopathic GBA1 wild-type PD (iPD) patients, and 5 GBA1-associated PD (PD-GBA) patients. GluCer/GalCer median ratios were found to be higher in the CSF of PD-GBA patients, particularly in severe GBA1 mutations, than those in iPD and HC. The observed trends in GluCer/GalCer ratios among groups provide novel information for the comprehensive analysis of sphingolipids as potential biomarkers of PD.

中文翻译：

液相色谱-串联质谱法分析帕金森病患者脑脊液中葡萄糖神经酰胺和半乳糖神经酰胺亚型的评价

编码葡萄糖脑苷脂酶 β 1 (GCase) 的 GBA1 突变是帕金森病 (PD) 最常见的遗传风险因素。 GCase 功能障碍导致葡萄糖神经酰胺 (GluCer) 底物在不同器官和体液中积聚。尽管量化生物样本中的 GluCer 同工型面临挑战，但它们作为 PD 生物标志物的潜在临床兴趣证明了开发稳健检测方法的合理性。提出了一种经过广泛评估的高效液相色谱-串联质谱 (HPLC-MS/MS) 方法，用于定量人脑脊液 (CSF) 样品中的 14 种 GluCer 和半乳糖神经酰胺 (GalCer) 亚型。优化了样品预处理、HPLC 和 MS/MS 参数。根据临床和实验室标准研究所和欧洲药品管理局指南的建议进行评估。生成了 4 条 7 点校准曲线，线性区间为 2.5 至 200 nM ( R ² ≥ 0.995)。定量限设定为 5 nM。运行间精密度和准确度分别高达 12.5% 和 9%。经过方法验证后，我们测量了 CSF 人类样本中 GluCer 和 GalCer 亚型的水平，包括 6 名健康对照 (HC)、22 名特发性 GBA1 野生型 PD (iPD) 患者和 5 名 GBA1 相关 PD (PD-GBA) 患者。研究发现，PD-GBA 患者的 CSF 中的 GluCer/GalCer 中值比率高于 iPD 和 HC 患者，尤其是严重的 GBA1 突变患者。观察到的各组间 GluCer/GalCer 比率的趋势为综合分析鞘脂作为 PD 的潜在生物标志物提供了新的信息。

更新日期：2024-07-24

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