Biological diagnosis of hemoglobin disorders is a complex process relying on the combination of several analytical techniques to identify Hb variants in a particular sample. Currently, hematology laboratories usually use high-performance liquid chromatography (HPLC), capillary electrophoresis and gel-based methods to characterize Hb variants. Co-elution and co-migration may represent major issues for precise identification of Hb variants, even for the most common ones such as Hb S and C. We adapted a top-down selected reaction monitoring (SRM) electron transfer dissociation (ETD) mass spectrometry (MS) method to fit with a clinical laboratory environment. An automated analytical process with semi-automated data analysis compatible with a clinical practice was developed. A comparative study between a reference HPLC method and the MS assay was performed on 152 patient samples. The developed workflow allowed to identify with high specificity and selectivity the most common Hb variants (Hb S and Hb C). Concordance of the MS-based approach with HPLC was 71/71 (100%) for Hb S and 11/11 (100%) for Hb C. This top-down SRM ETD method can be used in a clinical environment to detect Hb S and Hb C.

中文翻译：

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自上而下选择反应监测和电子转移解离对血红蛋白S和C鉴定的临床方法评价

血红蛋白疾病的生物学诊断是一个复杂的过程，它依赖于多种分析技术的组合来识别特定样本中的 Hb 变异体。目前，血液学实验室通常使用高效液相色谱 (HPLC)、毛细管电泳和基于凝胶的方法来表征 Hb 变异体。共洗脱和共迁移可能是精确识别 Hb 变体的主要问题，即使对于最常见的 Hb S 和 C 也是如此。我们采用了自上而下的选择反应监测 (SRM) 电子转移解离 (ETD) 质量光谱法 (MS) 方法以适应临床实验室环境。开发了一种与临床实践兼容的具有半自动化数据分析的自动化分析过程。对 152 个患者样本进行了参考 HPLC 方法和 MS 分析之间的比较研究。开发的工作流程允许以高特异性和选择性识别最常见的 Hb 变体（Hb S 和 Hb C）。基于 MS 的方法与 HPLC 的一致性为 Hb S 的 71/71 (100%) 和 Hb C 的 11/11 (100%)。这种自上而下的 SRM ETD 方法可用于临床环境中检测 Hb S和 Hb C。