• Title/Summary/Keyword: LC-MS/MS analysis

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Effect of Graviola Leaves Extracts on Antioxidant Activity and Melanin Production (멜라닌 생성 및 항산화 활성에 미치는 그라비올라 잎 추출물의 영향)

  • Lee, Kyuwon;Jang, Jiwon;Park, Sumin;Kang, Kihun;Yoon, Hyesoo;Ha, Yejin;Jeon, Sojeong;Ko, Hye Ju;Kim, Moon-Moo;Oh, Yunghee
    • Journal of Life Science
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    • v.29 no.6
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    • pp.662-670
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    • 2019
  • The purpose of this study was to investigate the effect of methanolic extracts of graviola, Annona muricate leaves (AMME) on antioxidant activity and melanin production. First of all, DPPH radical and reducing power were performed to determine the antioxidant effect of AMME and organic solvent fractions. AMME and organic solvent fractions showed antioxidative activity in a concentration dependent manner. The ethyl acetate fraction of AMME among organic solvent fractions showed the highest antioxidant activity. Moreover, tyrosinase activity was performed to confirm the effect of organic fractions on melanin production. AMME, ethyl acetate, and hexane fractions increased tyrosinase activity a dose dependent manner. Next, the hexane fraction with the best effect on melanin synthesis in AMME organic solvent fraction was divided into 12 fractions by silica column chromatography. Among them, the fraction 7 and 8 showed the highest DPPH radical scavenging activity and reducing power. In addition, the fraction 7 and 8 at $64{\mu}g/ml$ showed melanin synthesis by 260% and 184%, respectively. Finally, the fraction 8 at $4{\mu}g/ml$ showed melanin synthesis by 34% in B16F1 cells. LC-MS analysis showed that fraction 7 and fraction 8 have a molecular weight of 617 and 619, respectively. FT-IR analysis showed that fractions 7 and 8 is similar to bis(2-hydroxyethly)dimerate. Above results suggest that graviola leaves extracts could be applicable to the development of natural antioxidants or hair cosmetics which are related to the promoting effect of melanin production.

Cellular Protective Effect and Active Component Analysis of Lavender (Lavandula angustifolia) Extracts and Fractions (라벤더(Lavandula angustifolia) 추출물 및 분획물의 세포보호효과와 활성 성분 분석)

  • Kim, A Young;Ha, Ji Hoon;Kim, A Rang;Jeong, Hyo Jin;Kim, Kyoung Mi;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.28 no.4
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    • pp.479-484
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    • 2017
  • In this study, antioxidative activities and cellular protective effects of 70% ethanol extracts and fractions from lavender were evaluated. The scavenging activity ($FSC_{50}$) of free radical (1,1-phenyl-2-picrylhydrazyl, DPPH) was 46.6, 45.5 and $477.5{\mu}g/mL$ in the 70% ethanol extract, ethyl acetate fraction and aglycone fraction, respectively. The reactive oxygen species scavenging activities (${OSC_{50}$) of 70% ethanol extract, ethyl acetate fraction and aglycone fraction were 8.1, 3.3 and $17.6{\mu}g/mL$, respectively, and they showed lower antioxidative activity than that of using L-ascorbic acid ($1.5{\mu}g/mL$). However, the aglycone fraction showed higher photohemolysis protective effect than that of using the 70% ethanol extract and ethyl acetate fraction. At $50{\mu}M$ concentration, the cellular protective effect (${\tau}_{50}$) of 70% ethanol extract, ethyl acetate fraction and aglycone fraction from lavender was 70.6, 87.2 and 165.2 min, respectively. In particular, the lavender aglycone fraction showed 3.8 times higher cellular protective effect than that of (+)-${\alpha}$-tocopherol. The lavender fractional components including luteolin 7-O-glucuronide, vitextin, rosmarinic acid, luteolin, and apigenin were identified using TLC and LC-MS. However, the lavender aglycone fraction did not show any significant increase in flavonoids (luteolin and apigenin) compared to that of the ethyl acetate fraction. In conclusion, it is suggested that lavender may be applied as an antioxidant material in cosmetic industries.

Antioxidative and Cellular Protective Effects of Lysimachia christinae Hance Extract and Fractions (금전초 추출물 및 분획물의 항산화 활성 및 세포 보호 효과)

  • Kim, A Rang;Jung, Min Chul;Jeong, Hye In;Song, Dong Gi;Seo, Young Bin;Jeon, Young Hee;Park, So Hyun;Shin, Hyuk Soo;Lee, Sang Lae;Park, Soo Nam
    • Applied Chemistry for Engineering
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    • v.29 no.2
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    • pp.176-184
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    • 2018
  • In the present study, we investigated the antioxidative properties, cellular protective effects and component analyses of 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from Lysimachia christinae Hance (L. christinae Hance). In the evaluation of antioxidative properties, the free radical scavenging activities ($FSC_{50}$) of 50% ethanol extract, ethyl acetate fraction and aglycone fraction were 146.8, 22.2 and $27.2{\mu}g/mL$, respectively and total antioxidant capacities ($OSC_{50}$) were 29.3, 2.9 and $4.5{\mu}g/mL$, respectively. The ethyl acetate fraction showed the highest free radical scavenging activity and total antioxidant capacity. Also, the cellular protective effects (${\tau}_{50}$) of 50% ethanol extract, ethyl acetate fraction and aglycone fraction on $^1O_2$ induced photohemolysis of human erythrocytes were 26.9, 57.5 and 103.9 min at $5{\mu}g/mL$, respectively. In particular, ${\tau}_{50}$ of the aglycone fraction exhibited a higher cellular protective effect than that of (+)-${\alpha}$-tocopherol (37.7 min). The cell viability of the ethyl acetate fraction on the UVB-induced cell damage increased up to 90.1%. In addition, the ethyl acetate fraction ($5-25{\mu}g/mL$) showed cellular protective effects on the $H_2O_2-induced$ cell damages in a dose-dependent manner. TLC, HPLC, UV-vis spectroscopy and LC-MS were used to analyse components of the ethyl acetate fraction and the main components were quercetin, kaempferol and their glycosides. In conclusion, L. christinae Hance extract/fraction can function as antioxidants to protect the skin exposed to UV radiation and may also be used as a novel functional cosmetic material, for example, an antioxidant against skin photoaging.

Characteristics of Ginseng Extract and Its Effects on Sensory Properties of Pork Sausage (인삼 추출물의 사포닌 조성 특성과 추출물 첨가가 포크소시지 관능 특성에 미치는 영향)

  • Park, Kyoung-Mi;Hwang, In-Ho
    • Food Science of Animal Resources
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    • v.26 no.4
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    • pp.418-425
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    • 2006
  • This study was conducted to evaluate extraction properties of crude saponin and ginsenosides, and their effects on sensory properties of emulsified pork sausage. Non-dried ginseng root was boiled in 0 (e.g., 100% distilled water), 20, 40, 60, 80 or 100% ethanol, and powdered by a freezing dry method. Weight of dried powder for the 0% ethanol extraction was 20% of initial non-dried ginseng weight, while $20{\sim}80%$ and 100% ethanol extractions resulted in approximately 15 and 10% of their initial weights, respectively. On the other hand, crude saponin content in the dried powder was linearly increased for a higher ethanol content where 100% ethanol extraction resulted in 123.52 mg/g. LC/MS analysis of crude saponin for quantifying ginsenosides showed that Rb1, Rb2 and Rc were significantly (p<0.05) higher levels for both 80 and 100% ethanol extractions. In the case of Rg1 ginsenoside, 60, 80 and 100% ethanol extractions resulted in significantly (p<0.05) higher levels. Emulsified pork sausages containing 0, 1 or 2% ginseng extracts were smoked or non-smoked and their sensory characteristics and preference were evaluated. Smoking process significantly (p<0.05) decreased juiciness and tenderness, but the treatment significantly (p<0.05) improved flavor and consumer preference. It was particularly noticed that a 2% addition of ginseng extract prevented the adverse effects of smoking process on juiciness and tenderness while the 2% addition significantly (p<0.05) improved consumer preference. The current results implied that addition of ginseng extract in emulsified pork sausage could improve sensory quality.

Isolation and Characterization of Antifungal Compounds Produced by Bacillus polyfermenticus CJ6 Isolated from Meju (메주에서 분리한 Bacillus polyfermenticus CJ6가 생산하는 항진균 물질의 분리 및 특성)

  • Yang, Eun-Ju;Ma, Seung-Jin;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.57-65
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    • 2012
  • Antifungal compounds from Bacillus polyfermenticus CJ6 were purified using SPE, preparative HPLC, and reverse phase-HPLC. Antifungal compounds from B. polyfermenticus CJ6 were separated into three fractions (8, B, C) using preparative HPLC. LC/MS analysis of antifungal peaks suggested that B. polyfermenticus CJ6 produces lipopeptides; two kinds of iturin A ($C_{14}$, $C_{15}$), three kinds of surfactins ($C_{13}$, $C_{14}$, $C_{15}$), four kinds of fengycin A ($C_{14}$, $C_{15}$, $C_{16}$, $C_{17}$) and two kinds fengycin B ($C_{16}$, $C_{17}$). The antifungal activity of fraction 8, which was presumed as inturin A, was found to be stable after the pH, heat or proteolytic enzyme treatment, but it was unstable at 50-$70^{\circ}C$ for 24 hr. The antifungal activity of fraction B, which presumed as surfactins and fengycin A, was found to be stable after the heat treatment, but it was unstable in the pH 3.0 and after the protease (type I) or ${\alpha}$-chymotrypsin treatment. The antifungal activity of fraction C, which was presumed as fengycin A and B, was found to be stable in the pH 3.0-9.0 range and the heat treatment, but it was unstable with the treatment of protease (type I). The amino acid composition of the purified peaks 8-1 and 8-2 were Asx, Tyr, Gln, Pro, and Ser in a molar ratio of 3:1:1:1:1, which showed the same amino acid composition as iturin. From these results, we confirmed that antifungal compounds from B. polyfermenticus CJ6 most likely belonged to iturin A as well as surfactins and fengycins. As lipopeptides are known to act in a synergistic manner, the antifungal compounds from B. polyfermenticus CJ6 might have potential uses in biotechnology and biopharmaceutical applications.

Preventive Characteristics of Garlic Extracts Using in vitro Model System on Alzheimer's Disease (In vitro model system을 활용한 마늘 추출물의 치매예방 특성)

  • Choi, Gwi-Nam;Kim, Ji-Hye;Kwak, Ji-Hyun;Jeong, Chang-Ho;Jeong, Hee-Rok;Shin, Jung-Hye;Kang, Min-Jung;Sung, Nak-Ju;Heo, Ho-Jin
    • Journal of agriculture & life science
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    • v.44 no.4
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    • pp.45-55
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    • 2010
  • In this study, the acetylcholinesterase (AChE) inhibition and neuronal cell protective effects of water, 100% methanol and dichlromethane extracts from garlic were investigated. We found that dichloromethane extract of garlic resulted in a dose-dependent manner on AChE inhibition ($IC_{50}$: $36.1{\mu}g/mL$). In cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide (MTT), cell viabilities of water, 100% methanol and dichlromethane extracts were lower (almost under 40%) than amyloid ${\beta}$ protein ($A{\beta}$)-induced neurotoxicity. Because $A{\beta}$ is also known to increase neuronal cell membrane breakdown, neuronal apoptosis was further confirmed by lactate dehydrogenase (LDH) and neutral red uptake (NRU) assay. Water extract presented relative protection against $A{\beta}$-induced membrane damage in LDH assay. However all garlic extracts showed significant problem with decrease of cell viability in NRU assay, especially at dichloromethan extract. To determine active compounds in column fractions (98:2 fraction) from dichloromethane extract which showed significant AChE inhibitory effect, we performed HPLC and LC-MS analysis. It was supposed that garlic may contain allyl methyl disulfide, diallyl monosulfide, and diallyl disulfide as active compounds.

Studies on Antioxidant Activity and In Vitro Inhibitory Activity of Tyrosinase and Collagenase in Artocarpus nitidus subsp. lingnaensis (Merr.) F.M. Jarrett using 4 Parameter Logistic (변수 분석을 통한 아토카푸스 니티두스 추출물과 분획물의 항산화, 타이로시나제 및 콜라제나제 In Vitro 저해활성 연구)

  • Son, Kwang-Hee;Kim, Young Kook;Choi, Sangho;Zhang, Zhiyun;Shin, Dong-Ha;Lee, Jong Suk;Park, Ho-Yong
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.2
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    • pp.161-173
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    • 2019
  • In this study, the antioxidative and inhibitory activity of tyrosinase and collagenase for the solvent extract and silica column fractions of Artocarpus nitidus were evaluated. The activities were quantified using the 4 parameter logistic. LC/MS analysis showed that the major component of the fractions was polyphenol and the total polyphenol content of the extract was $48.1{\pm}2.6mg\;GAE/g$. The radical scavenging activities ($SC_{50}$) for 1,1-diphenyl-2-picrylhydrazyl of the extract, fraction-1 and fraction-2 were 16.7, 42.0 and $10.1{\mu}g/mL$, respectively. The value for fraction-2 was the closest to ascorbic acid ($1.5{\mu}g/mL$). The tyrosinase inhibitory activity of the extracts and the fractions showed $IC_{50}$ of 64.9, 0.9 and $1.2{\mu}g/mL$, respectively, and overall activity was higher than that of kojic acid ($7.4{\mu}g/mL$) and arbutin ($119.0{\mu}g/mL$). In the experiment by zebrafish embryo, the whitening activity of fraction-2 (27.5%) was higher than that of kojic acid (18.6%), and there was no adverse effect up to $500{\mu}g/mL$ of fraction-2. For the collagenase inhibitory activity, the samples showed $IC_{50}$ of 139.8, 20.6, and $16.8{\mu}g/mL$, respectively, which were competitive to 1, 10-Phenanthroline ($55.4{\mu}g/mL$). The extract and fraction-2 showed $IC_{50}$ of 61.8 and $67.1{\mu}g/mL$ for elastase. These results suggest that A. nitidus extract can be used as a cosmetic material useful for antioxidant, whitening, and prevention of skin aging without adverse effects.

Evaluation of Antioxidant, Cytoprotective and Antimicrobial Properties of Polygoni multiflori Radix Extract, Fractions and Its Major Constituent (하수오 추출물, 분획물 및 주성분의 항산화, 세포 보호 및 항균 활성에 관한 평가)

  • Shin, Hyuk Soo;Kim, Minwoo;Song, Jerry;Lee, Junseok;Ha, Yoonjeong;Jeon, Young Hee;Kim, Ji Woong;Lee, Yun Ju;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.4
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    • pp.407-417
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    • 2018
  • In this study, the antioxidant, cytoprotective and antimicrobial activities of 50% ethanol extract of Polygoni multiflori Radix (PMR) and its ethyl acetate fraction were evaluated to confirm the applicability as a functional ingredient. The activities of the major constituent of PMR were verified and 2, 3, 5, 4′-tetrahydroxystilbene 2-O-${\beta}$-D-glucoside (THSG) was confirmed to be the main component of extract and fraction using HPLC-DAD, LC-EIS-MS analysis. The phenolic and THSG contents of the ethyl acetate fraction were 11.1- and 3.0-folds higher than those of the ethanol extract, respectively. As a result of the DPPH assay and that of luminol dependent chemiluminescence assay in $Fe^{3+}$-EDTA/H2O2 system. the ethylacetate fraction was superior to the ethanol extract in free radical and ROS scavenging activities. Especially, the ethyl acetate fraction and THSG exhibited the similar scavenging activity like L-ascorbic acid in ROS scavenging activity. The ethyl acetate fraction perceived the most potent cytoprotective effect against oxidative damage of erythrocytes induced by photosensitization reaction, followed by the ethanol fraction, THSG and that of (+)-${\alpha}$-tocopherol, which was used as a positive control. Antimicrobial activities were evaluated by disc diffusion and broth microdilution assay against S. aureus, E. coli, P. aeruginosa and C. albicans. In particular, the antibacterial activity of the extract and fraction against S. aureus was superior to that of methyl paraben. Taken together, our results suggest that PMR could be used as a natural ingredient for antioxidant, cytoprotective and antimicrobial activities.

Comparative Analysis of Ginsenoside Content in Processed Red Ginseng Foods Based on Food Type and Formulation (홍삼가공식품의 식품유형별 및 제형별 진세노사이드 함량 비교)

  • Yun-Jeong Yi;Min-Su Chang;In-Sook Lee;Hyun-Jeong Kim;Hyun-Jeong Jang;In-Sook Hwang
    • Journal of Food Hygiene and Safety
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    • v.39 no.2
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    • pp.163-170
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    • 2024
  • Red ginseng is manufactured as a health-functional food and is also present in various food types and in different product forms. However, there is currently no standardized regulation of ginsenoside content in foods containing red ginseng. In the present study, we analyzed the ginsenoside content of 66 red ginseng-containing foods and 35 health-functional foods collected online and directly from the market. The ginsenoside content was assessed using liquid chromatography (LC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods. The ginsenoside content of the various food types ranged 0.0 (not detected)-71.567 mg per daily intake of foods containing red ginseng. Sugar-preserved foods had the highest ginsenoside content, followed by solid teas, liquid teas, and red ginseng beverages. For health-functional foods, the ginsenoside content ranged 3.4-58.5 mg per daily intake, with levels ranging 83-607% of the indicated amounts. All values met the established standards. Upon comparing red ginseng health-functional foods and red ginseng-containing foods, the average ginsenoside content was determined to be 18.21 and 8.79 mg, respectively, thus being nearly twice as high in health-functional foods. However, there was a minimal difference between the ginsenoside content of red and black ginseng, with values of 11.84 and 12.63 mg, respectively. These findings provide insights on the variations in ginsenoside content of red and black ginseng in various food forms. This information is expected to be valuable for future regulations and consumer choice of products containing red ginseng.

Target engagement of ginsenosides in mild cognitive impairment using mass spectrometry-based drug affinity responsive target stability

  • Zhu, Zhu;Li, Ruimei;Qin, Wei;Zhang, Hantao;Cheng, Yao;Chen, Feiyan;Chen, Cuihua;Chen, Lin;Zhao, Yunan
    • Journal of Ginseng Research
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    • v.46 no.6
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    • pp.750-758
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    • 2022
  • Background: Mild cognitive impairment (MCI) is a transitional condition between normality and dementia. Ginseng is known to have effects on attenuating cognitive deficits in neurogenerative diseases. Ginsenosides are the main bioactive component of ginseng, and their protein targets have not been fully understood. Furthermore, no thorough analysis is reported in ginsenoside-related protein targets in MCI. Methods: The candidate protein targets of ginsenosides in brain tissues were identified by drug affinity responsive target stability (DARTS) coupled with label-free liquid chromatography-mass spectrometry (LC-MS) analysis. Network pharmacology approach was used to collect the therapeutic targets for MCI. Based on the above-mentioned overlapping targets, we built up a proteineprotein interaction (PPI) network in STRING database and conducted gene ontology (GO) enrichment analysis. Finally, we assessed the effects of ginseng total saponins (GTS) and different ginsenosides on mitochondrial function by measuring the activity of the mitochondrial respiratory chain complex and performing molecular docking. Results: We screened 2526 MCI-related protein targets by databases and 349 ginsenoside-related protein targets by DARTS. On the basis of these 81 overlapping genes, enrichment analysis showed the mitochondria played an important role in GTS-mediated MCI pharmacological process. Mitochondrial function analysis showed GTS, protopanaxatriol (PPT), and Rd increased the activities of complex I in a dose-dependent manner. Molecular docking also predicted the docking pockets between PPT or Rd and mitochondrial respiratory chain complex I. Conclusion: This study indicated that ginsenosides might alleviate MCI by targeting respiratory chain complex I and regulating mitochondrial function, supporting ginseng's therapeutic application in cognitive deficits.