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A simple MALDI target plate with channel design to improve detection sensitivity and reproducibility for quantitative analysis of biomolecules

机译:一种简单的MALDI目标板,具有通道设计,提高了生物分子的定量分析的检测灵敏度和再现性

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摘要

Overcoming the detrimental effects of sweet spots during crystallization is an important step to improve the quantitative abilities of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. In this study, we introduce MALDI targets, which exhibit a channel design to reduce sweet spot phenomena and improve reproducibility. The size of the channels was 3.0 mm in length, 0.35 mm in depth, and 0.40 mm in width, adjusted to the width of the implemented laser beam. For sample deposition, the matrix/sample mixture was homogenously deposited into the channels using capillary action. To demonstrate the proof-of-principle, the novel plates were used for the quantification of acetyl-L-carnitine in human blood plasma using a combined standard addition and isotope dilution method. The results showed that the reproducibility of acetyl-L-carnitine detection was highly improved over a conventional MALDI-MS assay, with RSD values of less than 5.9% in comparison with 15.6% using the regular MALDI method. The limits of quantification using the new plates were lowered approximately two-fold in comparison with a standard rastering approach on a smooth stainless-steel plate. Matrix effects were also assessed and shown to be negligible. The new assay was subsequently applied to the quantification of acetyl-L-carnitine in human plasma samples.
机译:克服结晶期间甜点的不利影响是提高基质辅助激光解吸/电离(MALDI)质谱法的定量能力的重要步骤。在这项研究中,我们介绍了MALDI目标,该目标展示了渠道设计,以减少甜蜜点现象,提高再现性。通道的尺寸长度为3.0毫米,深度为0.35毫米,宽度为0.40mm,调节到实施的激光束的宽度。对于样品沉积,使用毛细管作用均匀地沉积基质/样品混合物进入通道中。为了证明原理证据,使用组合的标准添加和同位素稀释方法使用新颖的板用于量化人血浆中的乙酰-1-肉碱。结果表明,乙酰-1-肉碱检测的再现性在常规的MALDI-MS测定上高度改善,RSD值与使用常规MALDI方法相比,RSD值小于5.6%。与光滑的不锈钢板上的标准光栅化方法相比,使用新板的定量极限约为两倍。还评估了基质效应并显示可忽略不计。随后将新测定施用于人血浆样品中乙酰-1-肉碱的定量。

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