Contaminant Spot Check and Removal Assay (ContamSPOT) for Mass Spectrometry Analysis
Contaminant Spot Check and Removal Assay (ContamSPOT) for Mass Spectrometry Analysis
复制标题
DOI：
10.1021/acs.analchem.3c05020
复制
发表时间：
2024-01-31
影响因子：
7.4
通讯作者：
Hao，Ling
中科院分区：
文献类型：
--
作者：
Smeriglio，Noah;Li，Haorong;Hao，Ling
质谱（MS）分析经常受到来自洗涤剂、盐和聚合物的污染物的挑战，这些污染物会损害数据质量，并可能损坏色谱和MS仪器。然而，研究人员往往在获得数据后才发现污染问题。目前还没有灵敏度足以在MS分析之前从几微升样品中检测痕量污染物的污染物测定法。为了满足该领域的这一关键需求，我们开发了一种灵敏、快速且具有成本效益的污染物抽查和去除分析（ContamSPOT），用于检测和定量痕量污染物，如清洁剂、盐和MS样品制备工作流程中常用的其他化学品。在MS进样前仅使用1 μL样品，通过ContamSPOT比色法或荧光法在薄层色谱（TLC）板上定量污染物。我们还优化了污染物去除方法，以便在ContamSPOT显示阳性结果时以最小的损失挽救样品。然后，ContamSPOT被成功地应用于评估常用的自下而上的蛋白质组学方法的有效性，去除去污剂，肽回收，再现性和蛋白质组覆盖。我们预计ContamSPOT将被MS实验室广泛采用，作为MS注射前的最后一步质量检查点。我们提供了一个实用的决策树和一个逐步的协议与故障排除指南，以方便其他研究人员使用ContamSPOT。ContamSPOT还可以提供独特的样品清洁度读数，用于未来开发新的基于MS的样品制备方法。
Mass spectrometry (MS) analysis is often challenged by contaminations from detergents, salts, and polymers that compromise data quality and can damage the chromatography and MS instruments. However, researchers often discover contamination issues only after they acquire the data. There is no existing contaminant assay that is sensitive enough to detect trace amounts of contaminants from a few microliters of samples prior to MS analysis. To address this crucial need in the field, we developed a sensitive, rapid, and cost-effective contaminant spot check and removal assay (ContamSPOT) to detect and quantify trace amounts of contaminants, such as detergents, salts, and other chemicals commonly used in the MS sample preparation workflow. Only 1 μL of the sample was used prior to MS injection to quantify contaminants by ContamSPOT colorimetric or fluorometric assay on a thin layer chromatography (TLC) plate. We also optimized contaminant removal methods to salvage samples with minimal loss when ContamSPOT showed a positive result. ContamSPOT was then successfully applied to evaluate commonly used bottom-up proteomic methods regarding the effectiveness of removing detergent, peptide recovery, reproducibility, and proteome coverage. We expect ContamSPOT to be widely adopted by MS laboratories as a last-step quality checkpoint prior to MS injection. We provided a practical decision tree and a step-by-step protocol with a troubleshooting guide to facilitate the use of ContamSPOT by other researchers. ContamSPOT can also provide a unique readout of sample cleanliness for developing new MS-based sample preparation methods in the future.