Evaluation of different strategies for determination of selenomethionine (SeMet) in selenized yeast by asymmetrical flow field flow fractionation coupled to inductively coupled plasma mass spectrometry (AF4-ICP-MS).,Analytical Methods

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Evaluation of different strategies for determination of selenomethionine (SeMet) in selenized yeast by asymmetrical flow field flow fractionation coupled to inductively coupled plasma mass spectrometry (AF4-ICP-MS).
Analytical Methods ( IF 2.7 ) Pub Date : 2020-06-19 , DOI: 10.1039/d0ay00658k
Daniel B Alcântara _{1,

2,

3} , Ronaldo F Nascimento ₃ , Gisele S Lopes ₂ , Patricia Grinberg ₁

Affiliation

This manuscript exemplifies the prospective use of asymmetrical flow field flow fractionation (AF4) coupled to inductively coupled plasma mass spectrometry (ICP-MS) as a simple tool for chemical speciation of selenomethionine (SeMet) in selenized yeast. Several popular sample preparation methods were evaluated for their suitability to determine selenomethionine (SeMet) in selenized yeast by AF4-ICP-MS. These included water, methanesulfonic acid (MSA), formic acid (FA) and alkaline extractions. Alkaline extraction (using sodium dodecyl sulfate buffer) provided the best recovery/determination conditions for SeMet based on analysis of NRC certified reference material (CRM) SELM-1 since it minimized hydrolysis of the protein peptide bonds optimally required for the AF4 separation. The analytical performance of three different AF4 membranes (5, 10 and 500 kDa regenerated cellulose) was also evaluated. No significant difference in the recovery of SeMet was observed when using 5 and 10 kDa RC membranes, whereas the 500 kDa membrane resulted in a significant loss. The proposed method presents appropriate instrument and intra-assay precisions of 4.4–9.2% and 3.8% RSD, respectively, a detection limit of 0.49 μg L⁻¹ SeMet as Se and good linearity with correlation coefficients (R) between 0.996 – 0.999. This is the first report of use of AF4-ICP-MS for species specific quantitation of SeMet in selenized yeast demonstrating its efficient use as an alternative method to other traditional chromatographic techniques.

中文翻译：

通过不对称流场流动分离与电感耦合等离子体质谱法（AF4-ICP-MS）评估硒化酵母中硒代蛋氨酸（SeMet）的不同策略的评估。

该手稿举例说明了将不对称流场分馏（AF4）与电感耦合等离子体质谱（ICP-MS）耦合作为硒化酵母中硒代蛋氨酸（SeMet）化学形态分析的简单工具的预期用途。通过AF4-ICP-MS评估了几种流行的样品制备方法在确定硒化酵母中硒代蛋氨酸（SeMet）的适用性。其中包括水，甲磺酸（MSA），甲酸（FA）和碱提取液。基于NRC认证参考材料（CRM）SELM-1的分析，碱性提取（使用十二烷基硫酸钠缓冲液）为SeMet提供了最佳的回收/测定条件，因为它可以最大程度地减少AF4分离所需的蛋白质肽键的水解。三种不同的AF4膜的分析性能（5，还评估了10和500 kDa的再生纤维素。当使用5和10 kDa RC膜时，未观察到SeMet回收率的显着差异，而500 kDa RC膜导致了明显的损失。所提出的方法分别提供了适当的仪器和批内精密度，相对标准偏差为4.4–9.2％和3.8％，检测极限为0.49μgL^-1 SeMet作为Se且线性良好，相关系数（ R）在0.996-0.999之间。这是首次使用AF4-ICP-MS对硒化酵母中的SeMet进行物种特异性定量的报道，这表明它可以有效地替代其他传统色谱技术。

更新日期：2020-07-09

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