Development and validation of an ultra-performance liquid chromatography with tandem mass spectrometry method for determination of soluble repulsive guidance molecule A in human serum and cerebrospinal fluid.

IF 1.9 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS Bioanalysis Pub Date : 2024-01-01 Epub Date: 2024-10-10 DOI:10.1080/17576180.2024.2403241

Patrick Schreier, Lili Huang, Emma Fung, Jennifer Mollon, Bernhard Sielaff, Marc R Lake, Michael Schulz, Khader Awwad

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Abstract

Aim: Repulsive guidance molecule A (RGMa) is upregulated in neurodegenerative diseases. To assess RGMa levels in human serum and cerebrospinal fluid (CSF), a quantification method was developed and validated according to ICH M10 guideline.Methods & results: Sample preparation consisted of immunoprecipitation (IP, only for serum), digestion and purification followed by MS.Conclusion: An UPLC-MS/MS method was established and used to assess normal range of soluble RGMa levels in serum and CSF of healthy controls, and patients with mild cognitive impairment or Alzheimer's disease. The normal range was between 13.0-44.8 ng/ml (CSF) and 9.9-20.9 ng/ml (serum) in healthy controls. In the CSF of patients with mild cognitive impairment and Alzheimer's disease, total soluble RGMa was twofold lower while unchanged in serum.

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开发并验证测定人血清和脑脊液中可溶性斥导分子 A 的超高效液相色谱-串联质谱法。

目的：神经退行性疾病会导致斥导分子A（RGMa）上调。为了评估人血清和脑脊液（CSF）中的 RGMa 水平，我们根据 ICH M10 指南开发并验证了一种定量方法：样品制备包括免疫沉淀（IP，仅适用于血清）、消化和纯化，然后进行质谱分析：建立了一种UPLC-MS/MS方法，用于评估健康对照组、轻度认知障碍或阿尔茨海默病患者血清和脑脊液中可溶性RGMa水平的正常范围。健康对照组的正常范围为 13.0-44.8 纳克/毫升（脑脊液）和 9.9-20.9 纳克/毫升（血清）。在轻度认知障碍和阿尔茨海默病患者的脑脊液中，可溶性 RGMa 总量降低了两倍，而在血清中则保持不变。

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来源期刊

Bioanalysis BIOCHEMICAL RESEARCH METHODS-CHEMISTRY, ANALYTICAL

CiteScore

3.30

自引率

16.70%

发文量

审稿时长

2 months

期刊介绍： Reliable data obtained from selective, sensitive and reproducible analysis of xenobiotics and biotics in biological samples is a fundamental and crucial part of every successful drug development program. The same principles can also apply to many other areas of research such as forensic science, toxicology and sports doping testing. The bioanalytical field incorporates sophisticated techniques linking sample preparation and advanced separations with MS and NMR detection systems, automation and robotics. Standards set by regulatory bodies regarding method development and validation increasingly define the boundaries between speed and quality. Bioanalysis is a progressive discipline for which the future holds many exciting opportunities to further reduce sample volumes, analysis cost and environmental impact, as well as to improve sensitivity, specificity, accuracy, efficiency, assay throughput, data quality, data handling and processing. The journal Bioanalysis focuses on the techniques and methods used for the detection or quantitative study of analytes in human or animal biological samples. Bioanalysis encourages the submission of articles describing forward-looking applications, including biosensors, microfluidics, miniaturized analytical devices, and new hyphenated and multi-dimensional techniques. Bioanalysis delivers essential information in concise, at-a-glance article formats. Key advances in the field are reported and analyzed by international experts, providing an authoritative but accessible forum for the modern bioanalyst.