Pectolinarigenin, an aglycone of pectolinarin, has more potent inhibitory activities on melanogenesis than pectolinarin.

Pectolinarin and pectolinarigenin have been reported to be major compounds in Cirsium setidens. In the present study, we demonstrated inhibitory effects of pectolinarin and pectolinarigenin from C. setidens on melanogenesis. Melanin synthesis was decreased in both pectolinarin- and pectolinarigenin-treated melan-a cells and in a reconstructed human skin model. However, pectolinarigenin treatment showed more potent inhibitory activity of melanin synthesis than did pectolinarin treatment. The concentrations of pectolinarin and pectolinarigenin in C. setidens water extracts were determined by HPLC. Unfortunately, the amount of pectolinarigenin of C. setidens water extract was lower than that of pectolinarin. To increase the pectolinarigenin content in C. setidens water extract, several component conversion methods were studied. Consequently, we identified that microwave irradiation under 1% acetic acid was an optimum sugar elimination method.

Composition and Bioactivity of Lipophilic Metabolites from Needles and Twigs of Korean and Siberian Pines (Pinus koraiensis Siebold & Zucc. and Pinus sibirica Du Tour).

Lipophilic extractive metabolites in different parts of the shoot system (needles and defoliated twigs) of Korean pine, Pinus koraiensis, and Siberian pine, Pinus sibirica, were studied by GC/MS. Korean pine needles comprised mainly bornyl p-coumarate, heterocyclic 15-O-functionalized labdane type acids (lambertianic acid), 10-nonacosanol, sterols and their esters. While Siberian pine needles contained less bornyl p-coumarate, lambertianic acid, sterols and their esters, but were richer in other 15-O-functionalized labdane type acids. The major components of the twig extract of P. koraiensis were lambertianic acid, abietane and isopimarane type acids, cembrane type alcohols, 8-O-functionalized labdanoids, sterols, sterol esters, and acylglycerols. The same extract of P. sibirica differed in larger amounts of other 15-O-functionalized labdane type acids and pinolenic acid glycerides, but in less quantities of cembranoids and 8-O-functionalized labdanoids. The labdane type pinusolic acid was detected for the first time in Korean pine. P. koraiensis was found to be unique in the genus for an ability to synthesize phyllocladane diterpenoids. The content of bound Δ5 -unsaturated polymethylene-interrupted fatty acids in the twig extracts of the both pines was similar or superior to that in their seed oil. Among the pines' metabolites tested isocembrol was strongest in inhibition of both α-glucosidase (IC50 2.9 µg/ml) and NO production in activated macrophages (IC50 3.6 µg/ml).

Preventive and therapeutic effects of blueberry (Vaccinium corymbosum) extract against DSS-induced ulcerative colitis by regulation of antioxidant and inflammatory mediators.

Inflammatory bowel disease (IBD) is an inflammatory disorder caused by hyperactivation of effector immune cells that produce high levels of proinflammatory cytokines. The aims of our study were to determine whether orally administered blueberry extract (BE) could attenuate or prevent the development of experimental colitis in mice and to elucidate the mechanism of action. Female Balb/C mice (n=7) were randomized into groups differing in treatment conditions (prevention and treatment) and dose of BE (50 mg/kg body weight). Acute ulcerative colitis was induced by oral administration of 3% dextran sodium sulfate for 7 days in drinking water. Colonic mucosal injury was assessed by clinical, macroscopic, biochemical and histopathological examinations. BE significantly decreased disease activity index and improved the macroscopic and histological score of colons when compared to the colitis group (P<.05). BE markedly attenuated myeloperoxidase accumulation (colitis group 54.97±2.78 nmol/mg, treatment group 30.78±1.33 nmol/mg) and malondialdehyde in colon and prostaglandin E2 level in serum while increasing the levels of superoxide dismutase and catalase (colitis group 11.94±1.16 U/ml, BE treatment group 16.49±0.39 U/ml) compared with the colitis group (P<.05). mRNA levels of the cyclooxygenase (COX)-2, interferon-γ, interleukin (IL)-1ß and inducible nitric oxide synthase cytokines were determined by reverse transcriptase polymerase chain reaction. Immunohistochemical analysis showed that BE attenuates the expression of COX-2 and IL-1ß in colonic tissue. Moreover, BE reduced the nuclear translocation of nuclear transcription factor kappa B (NF-κB) by immunofluorescence analysis. Thus, the anti-inflammatory effect of BE at colorectal sites is a result of a number of mechanisms: antioxidation, down-regulation of the expression of inflammatory mediators and inhibition of the nuclear translocation of NF-κB.

Bioavailability of ginsenosides from white and red ginsengs in the simulated digestion model.

This study aims to investigate the bioavailability of ginsenosides during simulated digestion of white (WG) and red (RG) ginseng powders. Stability, bioaccessibility, and permeability of ginsenosides present in WG and RG were studied in a Caco-2 cell culture model coupled with oral, gastric, and small intestinal simulated digestion. Most ginsenosides in WG and RG were stable (>90%) during the simulated digestion. Bioaccessibilities of total ginsenosides during in vitro digestion of WG and RG were similar at approximately 85%. However, the bioaccessibility of protopanaxatriol type ginsenosides in the early food phase was greater than that of the protopanaxadiol type. The less polar RG ginsenosides were released later following the jejunum phase. Ginsenosides had low permeability (<1 × 10(-6) cm/s) through Caco-2 cell monolayers. These findings suggest that the WG and RG ginsenoside compositions affect bioaccessibility during digestion and that ginsenosides are poorly absorbed in humans.

Aceriphyllum rossii extract and its active compounds, quercetin and kaempferol inhibit IgE-mediated mast cell activation and passive cutaneous anaphylaxis.

Aceriphyllum rossii contains an abundant source of natural flavonoids with potential antioxidant, anticancer and anti-inflammatory properties. However, the effect of A. rossii extract (ARE) on immunoglobulin E(IgE)-mediated allergic responses remains unknown. In the present study, the effects of ARE and its active compounds, quercetin and kaempferol, on IgE-mediated rat basophilic leukemia mast cell activation and passive cutaneous anaphylaxis (PCA) were investigated. ARE, quercetin, and kaempferol inhibited secretion of ß-hexosaminidase and histamine, and reduced the production and mRNA expression of interleukin-4 and tumor necrosis factor-α. ARE also decreased the production of prostaglandin E2 and leukotriene B4 and expression of cyclooxygenase 2 and 5-lipoxygenase. Furthermore, ARE, quercetin, and kaempferol inhibited IgE-mediated phosphorylation of Syk, phospholipase Cγ, protein kinase C (PKC)µ, and the mitogen-activated protein kinases, extracellular signal-regulated kinase, p38, and c-Jun N-terminal kinase. ARE, quercetin, and kaempferol markedly suppressed mast cell-dependent PCA in IgE-sensitized mice. These results indicate that ARE and its active constituents, quercetin and kaempferol, may be a useful therapy for immediate-type hypersensitivity.

Optimisation of pressurised liquid extraction of antioxidants from black bamboo leaves.

To develop an efficient green extraction approach for recovering bioactive compounds from natural plants, the potential of using pressurised liquid extraction (PLE) was examined on black bamboo (Phyllostachys nigra) leaves, with ethanol/water as solvents. The superheated PLE process showed a higher recovery of most constituents and antioxidative activity, compared to reflux extraction, with a significantly improved recovery of the total phenolic (TP) and flavonoid (TF) content and DPPH radical scavenging ability. For a broad range of ethanol aqueous solutions and temperatures, 50% EtOH and 200°C (static time: 25min) gave the best performance, in terms of the TP and TF (75% EtOH) content yield and DPPH scavenging ability (25% EtOH). Under the optimised extraction conditions, eight main antioxidative compounds were isolated and identified with HPLC-ABTS(+) assay guidance and assessed for radical scavenging activity. The superheated extraction process for black bamboo leaves enhanced the antioxidant properties by increasing the extraction of the phenolic components.

Antioxidant activity of phenolics in leaves of three red pepper (Capsicum annuum) cultivars.

The antioxidant properties and phenolic profiles were first investigated in this paper on the leaves of three red pepper cultivars, Blackcuban (BCPL), Hongjinju (HPL), and Yeokgang-hongjanggun (YHPL). Of the ethanol extract of the three cultivars, BCPL showed potent antioxidant activities against the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and the 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical. Nine antioxidative compounds from the red pepper leaves were isolated and identified as one polyamine phenolic conjugate, N-caffeoylputrescine (1); three chlorogenic acid derivatives, 5-O-caffeoylquinic acid (2), 5-O-caffeoylquinic acid methyl ester (4), and 5-O-caffeoylquinic acid butyl ester (9); one anthocyanin, delphinidin-3-[4-trans-coumaroyl-l-rhamnosyl(1→6)glucopyranoside]-5-O-glucopyranoside (3); and four flavone glycosides, luteolin-7-O-apiofuranosyl(1→2)glucopyranoside (5), luteolin-7-O-glucopyranoside (6), apigenin 7-O-apiofuranosyl(1→2)glucopyranoside (7), apigenin-7-O-glucopyranoside (8). 1 and 3 had the greatest potential for radical-scavenging activity and HepG2 cells protecting effect against oxidative stress. BCPL exhibited the highest content of 1 and 3. Of the three cultivars BCPL may be considered a good source of antioxidants.

Structure, enzymatic transformation and anticancer activity of branched high molecular weight laminaran from brown alga Eisenia bicyclis.

The structure of high molecular weight laminaran from brown alga Eisenia bicyclis was investigated by chemical and enzymatic methods, NMR spectroscopy and mass spectrometry. The laminaran from E. bicyclis was characterized as 1,3;1,6-ß-D-glucan with the high content of 1,6-linked glucose residues (ratio of bonds 1,3:1,6=1.5:1), which are both in the branches and in the main chain of the laminaran. The degree of polymerization of fragments, building from 1,3-linked glucose residues with single glucose branches at C-6 or without it, was no more than four glucose residues. The main part of 1,3-linked glucose blocks was builded from disaccharide fragments. 1,6-Linked glucose residues were localized basically on non-reduced ends of molecules. The degree of polymerization of 1,6-linked blocks was not greater than three glucose residues. Laminaran contained laminarioligosaccharides, gentiobiose, gentiotriose and single glucose residues in the branches at the C-6. Laminaran and its products of enzymatic hydrolysis inhibited a colony formation of human melanoma SK-MEL-28 and colon cancer DLD-1 cells. It was shown that decreasing the molecular weight of native laminaran to a determined limit (degree of polymerization 9-23) and increasing the content of 1,6-linked glucose residues increased the anticancer effect. Therefore, they may be perspective antitumor agents.

Functional implication of ß-carotene hydroxylases in soybean nodulation.

Legume-Rhizobium spp. symbiosis requires signaling between the symbiotic partners and differential expression of plant genes during nodule development. Previously, we cloned a gene encoding a putative ß-carotene hydroxylase (GmBCH1) from soybean (Glycine max) whose expression increased during nodulation with Bradyrhizobium japonicum. In this work, we extended our study to three GmBCHs to examine their possible role(s) in nodule development, as they were additionally identified as nodule specific, along with the completion of the soybean genome. In situ hybridization revealed the expression of three GmBCHs (GmBCH1, GmBCH2, and GmBCH3) in the infected cells of root nodules, and their enzymatic activities were confirmed by functional assays in Escherichia coli. Localization of GmBCHs by transfecting Arabidopsis (Arabidopsis thaliana) protoplasts with green fluorescent protein fusions and by electron microscopic immunogold detection in soybean nodules indicated that GmBCH2 and GmBCH3 were present in plastids, while GmBCH1 appeared to be cytosolic. RNA interference of the GmBCHs severely impaired nitrogen fixation as well as nodule development. Surprisingly, we failed to detect zeaxanthin, a product of GmBCH, or any other carotenoids in nodules. Therefore, we examined the possibility that most of the carotenoids in nodules are converted or cleaved to other compounds. We detected the expression of some carotenoid cleavage dioxygenases (GmCCDs) in wild-type nodules and also a reduced amount of zeaxanthin in GmCCD8-expressing E. coli, suggesting cleavage of the carotenoid. In view of these findings, we propose that carotenoids such as zeaxanthin synthesized in root nodules are cleaved by GmCCDs, and we discuss the possible roles of the carotenoid cleavage products in nodulation.

Determination of major phlorotannins in Eisenia bicyclis using hydrophilic interaction chromatography: seasonal variation and extraction characteristics.

In this study, a hydrophilic interaction chromatography (HILIC) condition was developed for the simultaneous determination of five major phlorotannins from an extract of Eisenia bicyclis (Kjellman) Setchell with good linearity (r(2)>0.999). Based on this method, the seasonal variations and extraction characteristics, in terms of total extraction yield and the content of the phlorotannins, were investigated under various extraction conditions. In results, the yields and phlorotannins were increased two-to-four times in summer (June-October) and then, were decreased to normal levels in winter (November-March). In the extraction of E. bicyclis, ethanol percentage in water, extraction time and washing time significantly affected the yield of the extract and the phlorotannins, whereas the temperature and the sample/solvent ratio impacted the extraction to a lesser degree. These results will be useful information in the application of this macroalga in the commercial areas related to nutraceuticals, pharmaceuticals, and cosmeceuticals.

Chalcones from the flowers of Coreopsis lanceolata and their in vitro antioxidative activity.

Three new chalcones, 3,2'-dihydroxy-4,3'-dimethoxychalcone-4'-glucoside (1), 4'-O-(2'''-O-caffeoyl)2',3',3,4-tetrahydroxychalcone (2), and 2',4',3-trihydroxy-3',4-dimethoxychalcone (3), along with five known phenolics, were isolated from Coreopsis lanceolata flowers. The structures of the new compounds were elucidated by extensive spectroscopic methods including NMR and MS. The three new chalcones showed a good in vitro HepG2 cell protecting effect against tert-butylhydroperoxide-induced oxidative stress.

Lipophilic extracts from needles and defoliated twigs of Pinus pumila from two different populations.

Hexane extracts of needles and defoliated twigs of Pinus pumila (Pall.) Regel from two distant populations, located in the southwest and the east (i.e., Lake Baikal region and Sakhalin Island) of the species distribution range were studied by GC/MS analysis. Composition and retention indices of major components were determined. A drastic composition divergence for the extracts of P. pumila needles and defoliated twigs, depending on growth location, was established. Needle extracts from the eastern population sample contained mainly labdane-type acids (anticopalic acid derivatives), whereas the predominant components of needle extracts from the other population sample were abietane-type acids (abietic, neoabietic acids) and isopimarane-type diterpenoids (sandaracopimaric acid, sandaracopimaradien-3ß-ol). The main components of defoliated twig extracts from Sakhalin Island population sample were abietane-type acids and cembrane-type diterpenoids, while content of these compounds in the extracts of the southwestern marginal population sample was remarkably lower.

Low-volatile lipophilic compounds in needles, defoliated twigs, and outer bark of Pinus thunbergii.

Despite a long history of the use of Pinus thunbergii for technical, medicinal, agricultural, and other purposes, the composition of low-volatile metabolites in the used parts of the plant has been poorly investigated. We report here on the distribution of lipophilic extractive compounds in different parts of the shoot system (needles, defoliated twigs, outer bark) of P. thunbergii studied by GC/MS. The highest and lowest contents of lipophilic substances were found in defoliated twigs and in outer bark correspondingly. Acid compounds in the extract of needles comprised mainly labdane type diterpenoids (trans-communic acid), while in the extracts of defoliated twigs and outer bark the acids were represented predominantly by abietane type compounds (neoabietic, dehydroabietic, abietic, levopimaric and palustric acids). The major neutral components of the extract of needles were 10-nonacosanol, labdanoids (18-hydroxy-1 3-epi-manoyl oxide, trans-communol), and beta-sitosterol. In the case of the extract of defoliated twigs, labdanoids (18-hydroxy-13-epi-manoyl oxide, trans-communol, 13-epi-torulosol), serratane triterpenoids (3beta-methoxyserrat-14-en-21-one), and beta-sitosterol were the main neutral constituents, whereas serratanoids (3beta-methoxyserrat-14-en-21-one) alone dominated among the neutral compounds of the outer bark extract. Most of the neutral components and the labdane type acids were detected for the first time in organs and tissues of P. thunbergii. The distribution of lipophilic metabolites in the studied parts of P. thunbergii shoot system may be applied for chemotaxonomy purposes. Diversified accumulation of extractive substances in different organs of the plant should be taken into account for isolation of specific components from the pine raw material.

Study of skin anti-ageing and anti-inflammatory effects of dihydroquercetin, natural triterpenoids, and their synthetic derivatives.

Accessible triterpenoids of ursane and lupane series, the flavonoid dihydroquercetin and their synthetic derivatives with polar substituentss were tested in vitro for inhibition of collagenase 1 (MMP-1) in UVB irradiation assay. Ursolic acid and uvaol disuccinate were the most active inhibitors in the ursane series. In the lupane series, the best inhibition was manifested by carboxymethyl ester of betulonic acid and betulin succinates. Down- regulation of MMP-1 by dihydroquercetin and its synthetic derivatives surpassed the activity of a standard (retinoic acid).

Content of antioxidative caffeoylquinic acid derivatives in field-grown Ligularia fischeri (Ledeb.) Turcz and responses to sunlight.

Ligularia fischeri (Ledeb.) Turcz, a commercial leafy vegetable, contains caffeoylquinic acid derivatives (CQAs) as major phenolic constituents. The HPLC chromatograms of leaf extracts collected from different areas in Korea showed a significant variation in CQA amount, and two tri-O-caffeoylquinic acids (triCQAs) were purified and structurally identified by NMR and MS from this plant. Radical scavenging activities among CQAs were found to be increased in proportion to the number of caffeoyl groups. Since this plant prefers damp and shady growth conditions, the effects of sunlight were investigated by growing plantlets in sunlight and shade for four weeks. Greater leaf thickness and higher phenolic contents were found for leaves grown in sunlight than in shade. Four major CQAs-5-mono-O-caffeoylquinic acid (5-monoCQA), and 3,4-, 3,5-, and 4,5-di-O-caffeoylquinic acid (diCQA)-were induced by solar irradiation, whereas the content of these compounds decreased steadily in shade leaves. The leaves of L. fischeri clearly showed adaptation responses to sunlight, and these characteristics can be exploited for cultivation of this plant for potential use as a nutraceutical and functional food.

Edible seaweed, Eisenia bicyclis, protects retinal ganglion cells death caused by oxidative stress.

The purpose of the present study was to determine whether edible seaweed, Eisenia bicyclis, is effective in blunting the negative influence of N-methyl-D-aspartate (NMDA) on rat retinas and of oxidative stress-induced transformed retinal ganglion cell (RGC-5 cell line) death. The ethanol extract of E. bicyclis (EEEB) significantly attenuated the negative insult of L: -buthionine-(S,R)-sulfoximine plus glutamate on RGC-5 cells. Treatment of the RGC-5 cells with EEEB reduced the reactive oxygen species and recovered the reduced glutathione level caused by various radical species such as H(2)O(2), OH·, or O(2)·(-). Moreover, EEEB inhibited lipid peroxidation on rat brain homogenates caused by sodium nitroprusside. Applying NMDA to the retina affected the thickness of the inner plexiform layer (IPL) and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) produced a positive effect on ganglion cells. Importantly, EEEB protected the thinning of IPL and increased TUNEL positive cells in the ganglion cell layer (GCL). Five phlorotannin derivatives were isolated using chromatographic methods and liquid chromatography-mass spectroscopy analysis which has been known as an antioxidant. In conclusion, EEEB has a neuroprotective effect in vitro and in vivo. Furthermore, the major constituents of this extract, phlorotannins, could possibly be active compounds due to their antioxidative potency.

Anti-inflammatory activity of hydroxycinnamic acid derivatives isolated from corn bran in lipopolysaccharide-stimulated Raw 264.7 macrophages.

In this study, the effect of the 80% ethanolic extract of corn bran (EECB) on inhibition of nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells was investigated. The EECB inhibited LPS-induced NO production and iNOS expression in a dose-dependent manner. Four hydroxycinnamic acid derivatives (HADs), including two free cinnamic acids, p-coumaric acid (CA) and ferulic acid (FA), and their conjugate phenolic amides, p-dicoumaroyl-putrescine (DCP) and diferuloylputrescine (DFP), were found to be present in the EECB by LC-MS analysis, and DFP (378.66 µg/g) was the predominant phenolic compound, followed by DCP (7.83 µg/g)>CA (5.58 µg/g)>FA (1.84 µg/g). The four HADs significantly inhibited NO production and iNOS expression in a dose-dependent manner. Among the four HADs tested, DFP showed the most potent inhibition on NO production and iNOS mRNA and protein expression, followed by DCP>FA ≥ CA. DFP also exhibited the strongest inhibition on LPS-induced iNOS and NF-κB luciferase activity, which was followed by DCP ≥ FA (CA)>CA (FA). Thus, these results suggest that phenolic amides in the corn bran may be a potential source of natural anti-inflammatory agents.

A potential commercial source of fucoxanthin extracted from the microalga Phaeodactylum tricornutum.

Fucoxanthin, one of the main marine carotenoids, is abundant in macro- and microalgae. Here, fucoxanthin was isolated and structurally identified as the major carotenoid in the diatom Phaeodactylum tricornutum through chromatographic and spectroscopic methods, such as liquid chromatography-positive-ion atmospheric pressure chemical ionization/mass spectroscopy and nuclear magnetic resonance. This pigment was quantified by reverse-phase high-performance liquid chromatography, and a number of extraction procedures were assessed to investigate the effect of solvent type, extraction time, temperature, and extraction method (maceration, ultrasound-assisted extraction, Soxhlet extraction, and pressurized liquid extraction). Among the investigated solvents, ethanol provided the best fucoxanthin extraction yield (15.71 mg/g freeze-dried sample weight). Fucoxanthin content in the extracts produced by the different methods was quite constant (15.42-16.51 mg/g freeze-dried sample weight) but increased steeply based on the percentage of ethanol in water, emphasizing the importance of ethanol in the extraction. The results indicate that P. tricornutum is a rich source of fucoxanthin (at least ten times more abundant than that in macroalgae) that is easily extracted with ethanol, suggesting potential applications in human and animal food, health, and cosmetics.

Rapid identification and evaluation of antioxidant compounds from extracts of Petasites japonicus by hyphenated-HPLC techniques.

Gradient HPLC coupled to Diode Array Detector (DAD), MS/MS and NMR was applied to the rapid structure determination of major compounds of methanol extracts from leaves and roots of Petasites japonicus. The relative antioxidant capacities of the compounds were evaluated by an HPLC system with post-column on-line antioxidant detection based on 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid radical scavenging. Six compounds were successfully separated on a reverse-phase C(18) column and were identified as 5-caffeoylquinic acid (5-CQA), fukinolic acid (FA), 3,5-di-O-caffeoylquinic acid (3,5-DCQA), quercetin-3-O-(6″-acetyl)-ß-glucopyranoside (QAG), 4,5-di-O-caffeoylquinic acid (4,5-DCQA) and kaempferol-3-O-(6″-acetyl)-ß-glucopyranoside (KAG) by MS/MS and (1)H NMR data. Among these compounds, those containing a caffeoyl moiety (5-CQA, FA, 3,5- and 4,5-DCQA) showed relatively strong radical scavenging capacity, with 3,5-DCQA having the greatest radical scavenging capacity in leaf (23.09% of total antioxidant capacity) and root (26.47%) extracts. The relative radical scavenging portion of QAG was only 3.41% in the leaves and KAG did not show any radical scavenging activity. These results demonstrate that the hyphenated HPLC techniques can be successfully applied to rapidly identify structures and evaluate antioxidant activities without prior purification of compounds from plant tissues of P. japonicus.

Isolation of phlorotannins from Eisenia bicyclis and their hepatoprotective effect against oxidative stress induced by tert-butyl hyperoxide.

Eisenia bicyclis (Kjellman) Setchell is a common brown alga that inhabits the middle Pacific coast around Korea and Japan. In this study, the ethanol extract and its serial solvent fractions were prepared from fresh E. bicyclis, and their hepatoprotective effects were investigated against hepatotoxicity in tert-butyl hyperoxide(t-BHP)-injured HepG2 cells. When these samples were used at a dose of 10-40 µg/mL⁻¹, they significantly protected the t-BHP-induced cell death in HepG2 cells. Among fractions, ethyl acetate fraction (EF) and n-butanol extract (BF) exhibited potent hepatoprotective activities (62.60% for EF and 64.86% for BF) in t-BHP-injured HepG2 cells at a concentration of 10 µg/mL⁻¹. To find the potential factors for this activity, the samples were characterized on total phenolics, chlorophylls, carotenoids, and radical scavenging activity. Among them, EF showed the highest content of total phenolics and the strongest antioxidant activity both in on- and offline assays. Five phlorotannin compounds, oligomers of phloroglucinol, were isolated chromatographically from this fraction and structurally identified by (1)H-NMR and liquid chromatography-electrospray ionization-mass spectrometry analyses as eckol(1), 6,6'-bieckol(2), 8,8'-bieckol(3), dieckol(4), and phlorofucofuroeckol A(5). Compound 5 among five purified compounds showed the strongest protective activity (45.54%) at a concentration of 10 µM. At the high dose (40 µM), the protective activities of three compounds (compound 2, 4, and 5) were higher than that of quercetin treated with 10 µM concentration. Therefore, we can speculate that they can be developed as potential candidates for natural hepatoprotective agents.