Evaluation of a performic acid oxidation method for quantifying amino acids in freshwater species
Evaluation of a performic acid oxidation method for quantifying amino acids in freshwater species
复制标题
评估淡水物种中氨基酸定量的过甲酸氧化方法
DOI：
10.1002/lom3.10284
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
R. Bertolo
中科院分区：
文献类型：
--
作者：
Jennifer C. Thera;K. Kidd;Matthew G. Nosworthy;R. Bertolo
蛋白质中的巯基氨基酸储存汞等金属，但淡水物种中已建立的定量方法的应用和评估有限。因此，关于水生物种氨基酸组成的文献通常缺乏巯基半胱氨酸和甲硫氨酸。在这里，我们评估了过甲酸（PFA）氧化方法，以确定其适用于测量半胱氨酸和蛋氨酸，以及其他15种氨基酸，在新的矩阵（浮游动物，底栖无脊椎动物，鱼类和藻类）。用PFA氧化蛋白结合氨基酸，在盐酸中水解，用AccQ·Tagultra（沃茨）衍生化，并通过超高效液相色谱-荧光检测器分离。PFA氧化法成功地测定了15种氨基酸的精确结果，包括含硫氨基酸，酪氨酸（TYR）完全丢失，苯丙氨酸（PHE）的精确度较差。该方法的总体变异性为11%（不包括TYR和PHE），或以相对百分比报告时为6%，与其他基质中无PFA氧化的方法相当。除了TYR和硫醇，PFA氧化没有影响这些生物群的氨基酸组成。总的来说，研究结果是可重复的，与其他方法相当，并表明这是一个严格的方法来测量含硫氨基酸和水生生物群的整体氨基酸组成，这两者都是罕见的文献。
Thiol amino acids in proteins store metals like mercury, but established methods for their quantitation in freshwater species have had limited application and evaluation. As such, literature on the amino acid composition of aquatic species often lacks the thiols cysteine and methionine. Here, we evaluated a performic acid (PFA) oxidation method to determine its suitability to measure cysteine and methionine, as well as 15 other amino acids, in novel matrices (zooplankton, benthic invertebrates, fishes, and algae). Protein‐bound amino acids were oxidized with PFA, hydrolyzed in hydrochloric acid, derivatized with AccQ·Tagultra (Waters), and separated by ultra‐high performance liquid chromatography with fluorescence detection. PFA oxidation was successful in determining precise results for 15 amino acids, including the sulfur amino acids, with the complete loss of tyrosine (TYR) and poor precision of phenylalanine (PHE). The overall variability of the method was 11% (excluding TYR and PHE), or 6% when reported as relative percentages, comparable to methods without PFA oxidation in other matrices. Except for TYR and thiols, PFA oxidation did not affect the amino acid composition of these biota. Overall, the findings were reproducible and comparable to other approaches and suggest this is a rigorous method for measuring sulfur amino acids and the overall amino acid composition for aquatic biota, both of which are rare in the literature.
影响因子：
7.4
作者：
Armenta, Jenny M.;Cortes, Diego F.;Pisciotta, John M.;Shuman, Joel L.;Blakeslee, Kenneth;Rasoloson, Dominique;Ogunbiyi, Oluwatosin;Sullivan, David J., Jr.;Shulaev, Vladimir
通讯作者：
Shulaev, Vladimir