褐藻萃取物中間苯三酚衍生物之抗氧化物質能力分析及結構鑑定

Antioxidant activity analysis and structure identification of phloroglucinol derivatives from brown seaweed extracts

簡振育(CHEN-YU CHIEN)

指導教授 ： 黃郁慈

中原大學/工學院/化學工程學系/碩士(2023年)

https://doi.org/10.6840/cycu202301123

摘要

近年來因天然抗氧化劑可以降低因自由基所引起的發炎與器官退化、心血管疾病與減緩肌膚老化以及黑色素的生成，故受到大部分使用者的喜愛及重視。褐藻萃取物中含有間苯三酚衍生物，具備大量苯環與羥基，可藉由雙鍵共振特性吸收自由基達成抗氧化效果，屬於優良的天然抗氧化劑之一。
本研究建立一套溶劑萃取法萃取中國半葉馬尾藻 (Sargassum Hemiphyllum var. Chinense, S.H.v.c)，以及一系列的檢測方法並評估其抗氧化能力與抗黑色素沉澱能力，萃取結果顯示，S.H.v.c使用丙酮最佳萃取濃度為47.5%，萃取物含有總酚含量 (Total Phenolic Content, TPC) 24.86 mg 間苯三酚當量濃度 (Phloroglucinol Equivalents, PGE) / g、總黃酮含量 (Total Flavonoid Content, TFC) 14.92 mg 槲皮素當量濃度 (Quercetin Equivalents, QE) / g，抗氧化檢測中，鐵降低抗氧化能力檢測 (Ferric reducing antioxidant power, FRAP) 5.40 mMol Fe(Ⅱ) eq / g、總抗氧化檢測 (Total antioxidant capacity, TAC) 4.6 Trolox mM / g 和清除DPPH (1,1-diphenyl-2-picrylhydrazyl) 自由基檢測17.61% inhibition of Trolox，抗黑色素沉澱檢測中，酪胺酸酶抑制劑活性0.19 mU / g；使用乙醇最佳萃取濃度為38%，萃取物含有TPC：25.62 mg PGE / g、TFC：18.77 mg QE / g、FRAP：9.39 mMol Fe (Ⅱ) eq / g、TAC：9.7 Trolox mM / g、清除DPPH自由基檢測27.11% inhibition of Trolox與胺酸酶抑制劑活性0.129 mU / g。結果顯示S.H.v.c乙醇萃取物具有較佳抗氧化能力，而丙酮萃取物則具有較佳抗黑色素沉澱能力。
S.H.v.c萃取物藉由LC-QTOF質譜儀檢測，發現至少含有6種聚合度 (Degree of Polymerization, DP) 2 ~ 4之間苯三酚衍生物，如下所示：Phlorothols (DP=2)、Fucols (DP=2)、Eckol (DP=3)、Fuhalols (DP=3)、Fucophlorothols A (DP=3)，以及Tetrafucol A (DP=4)；另外藉由MALDI-TOF質譜儀檢測，鑑定出Dibenzo-p-dioxin-2,4,7,9-tetraol (DP=2)、Eckol (DP=3)、Fuhalos (DP=3)、Fucofuroeckol A (DP=3)與Diphlorethohydroxycarmalol (DP=4)，本研究使用上述LC-QTOF質譜儀與MALDI-TOF質譜儀檢測S.H.v.c萃取物中均鑑定出Eckol (C18H12O9，DP= 3，M.W.= 372 g/mol) 和 Fuhalols (C18H14O10，DP= 3，M.W.= 390 g/mol)，說明中國半葉馬尾藻富含聚合度較低的間苯三酚衍生物。

關鍵字

中國半葉馬尾藻 ； 間苯三酚 ； 總酚含量 ； 抗氧化能力檢測 ； 質譜儀檢測

並列摘要

In recent years, natural antioxidants can reduce inflammation and organ degradation caused by free radicals, cardiovascular diseases can also slow down skin aging and melanin production, so they are loved and valued by most users. The extract of brown algae contains phloroglucinol derivatives, which have a large number of benzene rings and hydroxyl groups. It can absorb free radicals by double bond resonance characteristics to achieve antioxidant effect. It is one of the excellent natural antioxidants.
In this study, a set of solvent extraction methods was established to extract Sargassum Hemiphyllum var. Chinese, S.H.v.c., and a series of detection methods were used to evaluate its antioxidant capacity and anti-melanin precipitation capacity. The extraction results showed that the best concentration of S.H.v.c using acetone was 47.5%, and the extract contained 24.86 mg of phloroglucinol equivalent (PGE)/g of total phenol content (TPC) Total flavone content (TFC) 14.92 mg quercetin equivalents (QE)/g, in antioxidant test, iron reduced antioxidant capacity test (FRAP) 5.40 mMol Fe (Ⅱ) eq/g, total antioxidant capacity test (TAC) 4.6 Trolox mM/g, DPPH free radical scavenging test 17.61% inhibition of Trolox, in anti melanin precipitation test, tyrosinase inhibitor activity 0.19 mU/g; The optimum extraction concentration of ethanol is 38%,and the extract contains TPC: 25.62 mg PGE/g, TFC: 18.77 mg QE/g, FRAP: 9.39 mMol Fe (Ⅱ) eq/g, TAC: 9.7 Trolox mM/g, DPPH free radical scavenging detection 27.11% inhibition of Trolox, and tyrosinase inhibitor activity 0.129 mU/g. The results showed that the ethanol extract of S.H.v.c had better antioxidant capacity, while the acetone extract had better anti melanin precipitation capacity.
The S.H.v.c extract was detected by LC-QTOF-MS and found to contain at least 6 degree of polymerization (DP) 2~4 phloroglucinol derivatives, as follows: Chlorothols (DP=2), Fucols (DP=2), Eckol (DP=3), Fuhalols (DP=3), Fucophothols A (DP=3), and Tetrafucol A (DP=4); In addition, by MALDI-TOF-MS detection, Dibenzo-p-dioxin-2,4,7,9-tetraol (DP=2), Eckol (DP=3), Fuhalos (DP=3), Fucofuroeckol A (DP=3) were identified and Diphlorethohydroxycarmalol (DP=4). Both analytical methods identified Eckol (C18H12O9，DP= 3，M.W.= 372 g/mol) and Fuhalols (C18H14O10，DP= 3，M.W.= 390 g/mol), indicating that Sargassum Hemiphyllum var. Chinese is rich in phloroglucinol derivatives with a low degree of polymerization.

並列關鍵字

Sargassum Hemiphyllum Var. Chinese ； Phloroglucinol ； Total Phenolic Content ； Antioxidant Test ； Mass Spectrometry Detect

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