Department of Medical Sciences, Clinical Chemistry, Uppsala University, Uppsala University Hospital, Entrance 61, 3rd Floor, Dag Hammarskjölds Väg 18, 751 85, Uppsala, Sweden.
Department of Clinical Sciences, Danderyd Hospital, Karolinska Institutet, Stockholm, Sweden.
Metabolomics. 2020 Feb 12;16(2):26. doi: 10.1007/s11306-020-1648-5.
Standardized commercial kits enable targeted metabolomics analysis and may thus provide an attractive complement to the more explorative approaches. The kits are typically developed for triple quadrupole mass spectrometers using serum and plasma.
Here we measure the concentrations of preselected metabolites in cerebrospinal fluid (CSF) using a kit developed for high-resolution mass spectrometry (HRMS). Secondarily, the study aimed to investigate metabolite alterations in patients with secondary progressive multiple sclerosis (SPMS) compared to controls.
We performed targeted metabolomics in human CSF on twelve SPMS patients and twelve age and sex-matched healthy controls using the Absolute IDQ-p400 kit (Biocrates Life Sciences AG) developed for HRMS. The extracts were analysed using two methods; liquid chromatography-mass spectrometry (LC-HRMS) and flow injection analysis-MS (FIA-HRMS).
Out of 408 targeted metabolites, 196 (48%) were detected above limit of detection and 35 were absolutely quantified. Metabolites analyzed using LC-HRMS had a median coefficient of variation (CV) of 3% and 2.5% between reinjections the same day and after prolonged storage, respectively. The corresponding results for the FIA-HRMS were a median CV of 27% and 21%, respectively. We found significantly (p < 0.05) elevated levels of glycine, asymmetric dimethylarginine (ADMA), glycerophospholipid PC-O (34:0) and sum of hexoses in SPMS patients compared to controls.
The Absolute IDQ-p400 kit could successfully be used for quantifying targeted metabolites in the CSF. Metabolites quantified using LC-HRMS showed superior reproducibility compared to FIA-HRMS.
标准化的商业试剂盒可实现靶向代谢组学分析，因此可能成为更具探索性方法的补充。这些试剂盒通常是针对使用血清和血浆的三重四极杆质谱仪开发的。
本研究旨在使用专为高分辨率质谱（HRMS）开发的试剂盒测量脑脊液（CSF）中预选代谢物的浓度。其次，研究旨在调查继发进展型多发性硬化症（SPMS）患者与对照组之间代谢物的改变。
我们使用专为 HRMS 开发的 Absolute IDQ-p400 试剂盒（Biocrates Life Sciences AG），对 12 名 SPMS 患者和 12 名年龄和性别匹配的健康对照者的人 CSF 进行靶向代谢组学分析。使用两种方法（液相色谱-高分辨质谱（LC-HRMS）和流动注射分析-MS（FIA-HRMS））分析提取物。
在 408 个靶向代谢物中，有 196 个（48%）检测到的浓度高于检测限，有 35 个代谢物绝对定量。使用 LC-HRMS 分析的代谢物在同一天重复注射和延长储存后的中值变异系数（CV）分别为 3%和 2.5%。FIA-HRMS 的相应结果分别为中值 CV 为 27%和 21%。与对照组相比，SPMS 患者的甘氨酸、非对称二甲基精氨酸（ADMA）、甘油磷脂 PC-O（34：0）和己糖总和的水平显著升高（p<0.05）。
Absolute IDQ-p400 试剂盒可成功用于 CSF 中靶向代谢物的定量。与 FIA-HRMS 相比，使用 LC-HRMS 定量的代谢物显示出更好的重现性。
