Critical Evaluation and Validation of a High-Throughput Microplate-Based Cupric Reducing Antioxidant Capacity Method for the Analysis of Fish Feed Ingredients.

IF 6 2区 医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Aleksander Arnø, Viviana Sarmiento, Odd Elvebø, Pedro Araujo
{"title":"Critical Evaluation and Validation of a High-Throughput Microplate-Based Cupric Reducing Antioxidant Capacity Method for the Analysis of Fish Feed Ingredients.","authors":"Aleksander Arnø, Viviana Sarmiento, Odd Elvebø, Pedro Araujo","doi":"10.3390/antiox14060728","DOIUrl":null,"url":null,"abstract":"<p><p>The cupric ion reducing antioxidant capacity (CUPRAC) assay, originally developed to measure the antioxidant capacity of nutritional products spectrophotometrically, utilized water as the solvent for Trolox. Due to the limited solubility of Trolox in aqueous solutions, the optimization of the solvent system was investigated to enhance analytical performance. Solvent combinations consisting of methanol, ethanol, and water were evaluated to identify the mixture that ensures complete dissolution and maximum absorbance signal, using a ternary plot diagram and mathematical modeling. A methanol/water ratio of 0.64:0.36 was identified as the optimal solvent composition. Under these conditions, the CUPRAC assay demonstrated a linear range of 0-50 μM, a limit of detection of 0.91 μM, and a limit of quantification of 2.75 μM. Precision, expressed as the coefficient of variation, was below 5%, and accuracy-defined as the deviation between nominal and back-calculated concentrations-remained within ±7.0%, in accordance with the variation range recommended by the International Committee on Harmonization. The estimated molar absorption coefficient at the optimized solvent ratio (ε<sub>Trolox</sub> = 2.62 × 10<sup>4</sup> L mol<sup>-1</sup> cm<sup>-1</sup>) was applied to determine the antioxidant capacity of fish commercial feed ingredients containing a mixture of rosemary and olive extracts.</p>","PeriodicalId":7984,"journal":{"name":"Antioxidants","volume":"14 6","pages":""},"PeriodicalIF":6.0000,"publicationDate":"2025-06-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12190020/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Antioxidants","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3390/antiox14060728","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

The cupric ion reducing antioxidant capacity (CUPRAC) assay, originally developed to measure the antioxidant capacity of nutritional products spectrophotometrically, utilized water as the solvent for Trolox. Due to the limited solubility of Trolox in aqueous solutions, the optimization of the solvent system was investigated to enhance analytical performance. Solvent combinations consisting of methanol, ethanol, and water were evaluated to identify the mixture that ensures complete dissolution and maximum absorbance signal, using a ternary plot diagram and mathematical modeling. A methanol/water ratio of 0.64:0.36 was identified as the optimal solvent composition. Under these conditions, the CUPRAC assay demonstrated a linear range of 0-50 μM, a limit of detection of 0.91 μM, and a limit of quantification of 2.75 μM. Precision, expressed as the coefficient of variation, was below 5%, and accuracy-defined as the deviation between nominal and back-calculated concentrations-remained within ±7.0%, in accordance with the variation range recommended by the International Committee on Harmonization. The estimated molar absorption coefficient at the optimized solvent ratio (εTrolox = 2.62 × 104 L mol-1 cm-1) was applied to determine the antioxidant capacity of fish commercial feed ingredients containing a mixture of rosemary and olive extracts.

基于微孔板的高通量铜还原抗氧化能力分析方法的关键评价和验证
铜离子还原抗氧化能力(CUPRAC)测定法最初是为了分光光度法测定营养品的抗氧化能力而开发的,它利用水作为Trolox的溶剂。由于Trolox在水溶液中的溶解度有限,为了提高分析性能,对溶剂体系进行了优化研究。使用三元图和数学建模,评估了由甲醇、乙醇和水组成的溶剂组合,以确定确保完全溶解和最大吸光度信号的混合物。优选甲醇/水比为0.64:0.36。在此条件下,CUPRAC的线性范围为0 ~ 50 μM,检测限为0.91 μM,定量限为2.75 μM。精密度(以变异系数表示)低于5%,准确度(定义为标称浓度与背算浓度之间的偏差)保持在±7.0%以内,符合国际协调委员会推荐的变异范围。利用最佳溶剂比(εTrolox = 2.62 × 104 L mol-1 cm-1)下的摩尔吸收系数测定了迷迭香和橄榄提取物混合饲料的抗氧化能力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Antioxidants
Antioxidants Biochemistry, Genetics and Molecular Biology-Physiology
CiteScore
10.60
自引率
11.40%
发文量
2123
审稿时长
16.3 days
期刊介绍: Antioxidants (ISSN 2076-3921), provides an advanced forum for studies related to the science and technology of antioxidants. It publishes research papers, reviews and communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Electronic files and software regarding the full details of the calculation or experimental procedure, if unable to be published in a normal way, can be deposited as supplementary electronic material.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信