Qualification of a surrogate matrix-based absolute quantification method for amyloid-β₄₂ in human cerebrospinal fluid using 2D UPLC-tandem mass spectrometry.
Qualification of a surrogate matrix-based absolute quantification method for amyloid-β₄₂ in human cerebrospinal fluid using 2D UPLC-tandem mass spectrometry.
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DOI：
10.3233/jad-132489
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Shaw LM
中科院分区：
文献类型：
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作者：
Korecka M;Waligorska T;Figurski M;Toledo JB;Arnold SE;Grossman M;Trojanowski JQ;Shaw LM
这项工作的主要目的是：1)建立一个校准替代基质，用于定量测定人脑脊液（CSF）中的淀粉样蛋白-β (a β)42，并为LC-MS-MS方法制备质量控制样品；2)验证该方法的分析性能；3)评估其诊断效用，并将其与AlzBio3免疫分析法进行比较。分析方法基于2D-UPLC-MS-MS平台。样品前处理采用5 M盐酸胍，采用μ洗脱固相萃取柱进行萃取。柱清洗过程包括乙腈逐步去除水溶液，确保长期色谱性能一致。接受者-操作者特征（ROC）曲线和相关性分析评估了UPLC-MS-MS与AlzBio3免疫分析法在检测阿尔茨海默病（AD）方面的诊断效用。替代基质，含4mg /mL牛血清白蛋白的人工脑脊液，可提供线性和可重复性校准，可与作为校准基质的人合并脑脊液相比较。每天对捕获柱和分析柱进行适当的清洁，无故障运行。脑脊液Aβ42的分析表明，UPLC-MS-MS将神经病理诊断的AD受试者与健康对照区分开来，其诊断效用至少与AlzBio3相当。两种方法的ROC曲线比较无统计学差异（p = 0.2229）。线性回归分析表明，与AlzBio3免疫分析法相比，该质谱法测定的Aβ42浓度显著升高，但结果高度相关。综上所述，新建立的2D-UPLC-MS-MS测定Aβ42的替代基质具有选择性、重复性和准确性。记录的AD与对照的分析性能和诊断性能支持考虑作为候选参考方法。
The primary aims of this work were to: 1) establish a calibrator surrogate matrix for quantification of amyloid-β (Aβ)42 in human cerebrospinal fluid (CSF) and preparation of quality control samples for LC-MS-MS methodology, 2) validate analytical performance of the assay, and 3) evaluate its diagnostic utility and compare it with the AlzBio3 immunoassay. The analytical methodology was based on a 2D-UPLC-MS-MS platform. Sample pretreatment used 5 M guanidine hydrochloride and extraction on μElution SPE columns as previously described. A column cleaning procedure involved gradual removal of aqueous solvents by acetonitrile assured consistent long-term chromatography performance. Receiver-operator characteristic (ROC) curve and correlation analyses evaluated the diagnostic utility of UPLC-MS-MS compared to AlzBio3 immunoassay for detection of Alzheimer’s disease (AD). The surrogate matrix, artificial CSF containing 4 mg/mL of BSA, provides linear and reproducible calibration comparable to human pooled CSF as calibration matrix. Appropriate cleaning of the trapping and analytical columns provided every-day, trouble-free runs. Analyses of CSF Aβ42 showed that UPLC-MS-MS distinguished neuropathologically-diagnosed AD subjects from healthy controls with at least equivalent diagnostic utility to AlzBio3. Comparison of ROC curves for these two assays showed no statistically significant difference (p = 0.2229). Linear regression analysis of Aβ42 concentrations measured by this mass spectrometry-based method compared to the AlzBio3 immunoassay showed significantly higher but highly correlated results. In conclusion, the newly established surrogate matrix for 2D-UPLC-MS-MS measurement of Aβ42 provides selective, reproducible, and accurate results. The documented analytical performance and diagnostic performance for AD versus controls supports consideration as a candidate reference method.