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首页> 外文期刊>Bioanalysis >Development of an UPLC/MS–MS method for quantification of intact IGF-I from human serum
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Development of an UPLC/MS–MS method for quantification of intact IGF-I from human serum

机译：用于从人血清定量完整IGF-1的UPLC / MS-MS方法的开发

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Aim: Developing LC–MS methods for biomolecules is often challenging due to issues with molecular size and complexity, nonspecific binding, protein binding, solubility and sensitivity. As a result, complex sample preparation workflows, including immune-affinity and/or protein digestion and lengthy analysis potentially using nano-flow LC, may be needed to achieve the required sensitivity. This work aims to provide a simple, sensitive, fast and robust method for quantification of intact IGF-I from human serum using UPLC–MS/MS. Methods: IGF-I serum samples were denatured with sodium dodecyl sulfate, followed by organic protein precipitation to effectively disrupt protein binding and subsequent SPE of the resulting supernatant for sample cleanup and enrichment prior to LC–MS/MS analysis. Separation was performed on an analytical scale LC using a reversed-phase column containing 0.99) and mean accuracy of 101.76%. Accuracies for quality control samples were between 93.9–107.7% with RSD <7%. Conclusion: The analytical sensitivity, linear dynamic range and excellent reproducibility of this method reliably measures endogenous and elevated serum IGF-I levels, demonstrating its utility in discovery, bioanalysis and clinical research.
机译：目的：由于分子尺寸和复杂性，非特异性结合，蛋白质结合，溶解度和敏感性，开发用于生物分子的LC-MS方法通常是具有挑战性的。结果，可能需要复合的样品制备工作流程，包括使用纳米流动LC可能使用纳米流动LC的免疫亲和力和/或蛋白质消化和冗长的分析，以达到所需的灵敏度。这项工作旨在提供使用UPLC-MS / MS的来自人血清的完整IGF-I的简单，灵敏，快速和坚固的方法。方法：IGF-1血清样品用十二烷基硫酸钠变性，然后用有机蛋白质沉淀，以有效地破坏所得上清液的蛋白质结合和随后的样品清洁剂，并在LC-MS / MS分析之前进行富集。使用含有0.99）的反相色谱柱在分析尺度LC上进行分离，平均精度为101.76％。质量控制样品的精度在93.9-107.7％之间，RSD <7％。结论：分析敏感性，线性动态范围和该方法的优异再现性可靠地测量内源性和升高的血清IGF-I水平，展示其在发现，生物分析和临床研究中的效用。

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