{"title":"快速和成本效益的微孔板测定法准确定量的总酚在海藻","authors":"Rosalía Martínez-López, Maria G. Tuohy","doi":"10.1016/j.fochms.2023.100166","DOIUrl":null,"url":null,"abstract":"<div><p>Brown seaweeds (<em>Phaeophyceae</em>) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R<sup>2</sup>) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from <em>A. nodosum</em> demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2–3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format.</p></div>","PeriodicalId":34477,"journal":{"name":"Food Chemistry Molecular Sciences","volume":"6 ","pages":"Article 100166"},"PeriodicalIF":4.1000,"publicationDate":"2023-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/11/51/main.PMC9982613.pdf","citationCount":"1","resultStr":"{\"title\":\"Rapid and cost-efficient microplate assay for the accurate quantification of total phenolics in seaweeds\",\"authors\":\"Rosalía Martínez-López, Maria G. Tuohy\",\"doi\":\"10.1016/j.fochms.2023.100166\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Brown seaweeds (<em>Phaeophyceae</em>) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R<sup>2</sup>) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from <em>A. nodosum</em> demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2–3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format.</p></div>\",\"PeriodicalId\":34477,\"journal\":{\"name\":\"Food Chemistry Molecular Sciences\",\"volume\":\"6 \",\"pages\":\"Article 100166\"},\"PeriodicalIF\":4.1000,\"publicationDate\":\"2023-07-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/11/51/main.PMC9982613.pdf\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Food Chemistry Molecular Sciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2666566223000060\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"FOOD SCIENCE & TECHNOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Food Chemistry Molecular Sciences","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2666566223000060","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"FOOD SCIENCE & TECHNOLOGY","Score":null,"Total":0}
引用次数: 1
摘要
褐海藻(Phaeophyceae)是多酚的丰富来源(高达20%的干重)，其结构基于间苯三酚(1,3,5-三羟基苯)。迄今为止，总酚含量(TPC)的测定涉及与Folin-Ciocalteu (FC)试剂的氧化还原反应。然而，与其他还原性物质的副反应妨碍了TPC的准确、直接测量。本研究报告了一种新的微孔板检测方法，涉及间苯三酚与Fast Blue BB (FBBB)重氮盐在碱性pH下的偶联反应，形成稳定的三偶氮络合物，最大吸光度在450 nm处。以间苯三酚为标准，线性回归相关值(R2)≥0.99。对紫草粗水提取物和乙醇提取物中间苯三酚当量(PGEs)的直接定量分析表明，新的FBBB分析不受侧氧化还原干扰，并且在相对快速(30分钟)，成本效益(0.24欧元/次)的微孔板格式下提供了更准确的TPC估计(比FC分析低1.2 - 3.9倍)。
Rapid and cost-efficient microplate assay for the accurate quantification of total phenolics in seaweeds
Brown seaweeds (Phaeophyceae) are a rich source of polyphenols (up to 20% dry weight) with a structure based on phloroglucinol (1,3,5-trihydroxybenzene). To-date the determination of total phenolics content (TPC) involves a redox reaction with the Folin-Ciocalteu (FC) reagent. However, side reactions with other reducing substances preclude accurate, direct measurement of TPC. This research reports a novel microplate assay involving a coupling reaction between phloroglucinol with Fast Blue BB (FBBB) diazonium salt, at basic pH, to form a stable tri-azo complex with maximum absorbance at 450 nm. Linear regression correlation values (R2) were ≥0.99 with phloroglucinol as standard. Direct quantification of TPCs (phloroglucinol equivalents, PGEs) in crude aqueous and ethanolic extracts from A. nodosum demonstrated that the new FBBB assay is not subject to side-redox interference and provides a more accurate estimate of TPC (1.2–3.9-fold lower than with the FC assay) in a relatively rapid (30 min), cost-effective (0.24€/test) microplate format.
期刊介绍：
Food Chemistry: Molecular Sciences is one of three companion journals to the highly respected Food Chemistry.
Food Chemistry: Molecular Sciences is an open access journal publishing research advancing the theory and practice of molecular sciences of foods.
The types of articles considered are original research articles, analytical methods, comprehensive reviews and commentaries.
Topics include:
Molecular sciences relating to major and minor components of food (nutrients and bioactives) and their physiological, sensory, flavour, and microbiological aspects; data must be sufficient to demonstrate relevance to foods and as consumed by humans
Changes in molecular composition or structure in foods occurring or induced during growth, distribution and processing (industrial or domestic) or as a result of human metabolism
Quality, safety, authenticity and traceability of foods and packaging materials
Valorisation of food waste arising from processing and exploitation of by-products
Molecular sciences of additives, contaminants including agro-chemicals, together with their metabolism, food fate and benefit: risk to human health
Novel analytical and computational (bioinformatics) methods related to foods as consumed, nutrients and bioactives, sensory, metabolic fate, and origins of foods. Articles must be concerned with new or novel methods or novel uses and must be applied to real-world samples to demonstrate robustness. Those dealing with significant improvements to existing methods or foods and commodities from different regions, and re-use of existing data will be considered, provided authors can establish sufficient originality.
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