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首页> 外文期刊>Analytica chimica acta >Measurement of uncertainty in peptide molecular weight determination using size-exclusion chromatography with multi-angle laser light-scattering detection and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
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Measurement of uncertainty in peptide molecular weight determination using size-exclusion chromatography with multi-angle laser light-scattering detection and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

机译：使用尺寸排阻色谱法，多角度激光散射检测和基质辅助激光解吸/电离飞行时间质谱法测定肽分子量中的不确定性

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摘要

The molecular weight of biopolymers such as peptides or proteins is vital information for understanding their physical/chemical properties.The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS)and size-exclusion chromotography-multi-angle laser light scattering(SEC-MALLS)techniques each with its advantages and limitations were used for molecular weight determination of the SPf66 peptide.The precision of each method was studied using a two-factor fully-nested design with all the analyses performed by the same operator on a single instrument.The overall precision corresponded to the time-different intermediate precision(i.e.time and repeatability conditions).In the case of SEC-MALLS,all potential uncertainty components were carefully evaluated in an Ishikawa diagram,then included and mathematically combined with the uncertainty arising from the accuracy assessment to provide the overall uncertainty.In this case,the refractive-index increment with solute concentration value(dn/dc)provided the most significant contribution to the combined uncertainty.A method for its quantitation is proposed.The accuracy of method B(SEC-MALLS)against reference method A(MALDI-TOF-MS)was assessed using the interval testing hypothesis to limit the risk of unacceptable bias.The results indicate that the bias of B is higher than the limit established at 5%,and is therefore not traceable to A under the studied conditions.
机译：诸如肽或蛋白质之类的生物聚合物的分子量对于了解其物理/化学性质至关重要。基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）和体积排阻色谱-多角散射光技术（SEC-MALLS）各自具有优势和局限性，可用于测定SPf66肽的分子量。采用两因素全嵌套设计对每种方法的精度进行了研究，所有分析均通过整体精度对应于时差中间精度（即时间和重复性条件）。对于SEC-MALLS，在Ishikawa图中仔细评估了所有潜在不确定性分量，然后包括在数学上结合精度评估带来的不确定性来提供整体不确定性。在这种情况下，折射率增量随s溶质浓度值（dn / dc）对组合不确定度的影响最大。提出了一种定量方法。采用参考方法A（MALDI-TOF-MS）评估了方法B（SEC-MALLS）的准确性。结果表明，B的偏差高于5％的上限，因此在研究条件下无法追溯到A。

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