• Title/Summary/Keyword: High performance Liquid Chromatography

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Validation of HPLC Methods for Ascorbic Acid and Its Derivatives in Foods (식품 중 아스코르빈산 유래 산화방지제의 HPLC 분석법 검증 및 개선)

  • Jeong, Min Kyu;Park, Chan Uk;Park, Min Hee;Yeo, JuDong;Park, SeungKwan;Kim, SoHee;Shin, Tae-Sun;Baek, Hyung Hee;Lee, JaeHwan
    • Food Engineering Progress
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    • v.15 no.1
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    • pp.75-79
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    • 2011
  • Analytical methods for food antioxidants including ascorbic acid, erythorbic acid, ascorbyl palmitate (AP), and ascorbyl stearate (AS), were validated using high performance liquid chromatography. Validation parameters such as linearity, limit of detection (LOD), limit of quantification (LOQ), and recovery were tested using lard and cider as food model systems. Linearity of ascorbic acid and erythorbic acid were both higher than ($R^2$> 0.99), LOD of these compounds were 0.46 and 0.48 ${\mu}g/mL$, respectively and LOQ were 1.39 and 1.45 ${\mu}g/mL$, respectively. The recovery rates of these compounds were 86.35-94.78% and 84.76-95.02%, respectively. However, the concentration of AP and AS decreased in methanol stock solution. Four other solvents including ethanol, acetonitrile, mixture of methanol and acetonitrile, and mixture of ethanol and acetonitrile were tested to increase the stability of AP and AS under room temperature and refrigerated temperature. Ethanol provided better stability of AP and AS under both room and refrigerated temperature. This study can help to accurately analyze the content of ascorbic acid and its derivatives in processed foods.

Anti-inflammatory effects of ethyl acetate fraction of unripe astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) on lipopolysaccharide-stimulated RAW 264.7 cells (지방질다당류로 자극한 RAW 264.7 세포에서 청도반시 땡감 에틸 아세테이트 분획물의 항염증 효과)

  • Park, Ye Bin;Jeong, Ha-Ram;Lee, Seung Hwan;Kim, Taewan;Kim, Dae-Ok
    • Korean Journal of Food Science and Technology
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    • v.51 no.1
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    • pp.90-96
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    • 2019
  • Unripe astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) is a by-product produced when thinning out the superfluous fruit of persimmon. We investigated whether unripe astringent persimmon has antioxidative and anti-inflammatory effects. Unripe astringent persimmon extract was fractionated sequentially in n-hexane, chloroform, ethyl acetate, n-butanol, and water. The ethyl acetate fraction had the highest total phenolic content, total flavonoid content, and antioxidant capacity compared to those of the other fractions. Pretreatment of lipopolysaccharide-stimulated RAW 264.7 macrophages with the ethyl acetate fraction reduced nitric oxide, interleukin-6, and intracellular oxidative stress in a dose-dependent manner. Ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry analysis revealed gallic acid, protocatechuic acid, 4-hydroxybenzoic acid, quercetin-3-O-glucoside, quercetin, and p-coumaric acid as the phenolic compounds of the ethyl acetate fraction. Collectively, these findings suggest that unripe astringent persimmon is a source of functional materials that can promote antioxidative and anti-inflammatory effects.

Concurrent Innate Immunity Activation and Anti-inflammation effects of Dialyzed Coffee Extract in RAW 264.7 Cells, Murine Macrophage Lineage (RAW 264.7 세포에서 투석시킨 커피 추출액의 선천면역활성화와 항염증의 동시발생)

  • Yoon, Cheol Soo;Lee, Suk Keun
    • The Korean Journal of Oral and Maxillofacial Pathology
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    • v.41 no.3
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    • pp.121-129
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    • 2017
  • Coffee (Coffea spp.) is one of the most important agricultural commodities, being widely consumed in the world. Various beneficial health effects of coffee have been extensively investigated, but data on habitual coffee consumption and its bio-physiological effect have not been clearly explained as well as it is not proved the cause and effect between drinking coffee and its bio-physiological reactions. We made the dialyzed coffee extract (DCE), which is absorbable through gastrointestinal tract, in order to elucidate the cellular effect of whole small coffee molecules. RAW 264.7 cells, a murine macrophage lineage, were directly treated with DCE, i.e., DCE-2.5 (equivalent to 2.5 cups of coffee a day), DCE-5, and DCE-10, for 12 hours, and their protein extracts were examined by immunoprecipitation high performance liquid chromatography (IP-HPLC). RAW 264.7 cells differently expressed the inflammation-related proteins depending on the doses of DCE. RAW 264.7 cells treated with DCE showed marked increase of cathepsin C, cathepsin G, CD20, CD28, CD31, CD68, indicating the activation of innate immunity. Particularly, the macrophage biomarkers, cathepsin G, cathepsin C, CD31, and CD68 were markedly increased after DCE-5 and DCE-10 treatments, and the lymphocyte biomarkers, CD20 and CD28 were consistently increased and became marked after DCE-10 treatment. On the other hand, RAW 264.7 cells treated with DCE showed consistent increase of IL-10, an anti-inflammatory factor, but gradual decreases of different pro-inflammatory proteins including $TNF{\alpha}$, COX-2, lysozyme, MMP-2, and MMP-3. In particular, the cellular signaling of inflammation was gradually mitigated by the reduction of $TNF{\alpha}$, COX-2, IL-12, and M-CSF, and also the matrix inflammatory reaction was reduced by marked deceases of MMP-2, MMP-3, and lysozyme. These anti-inflammatory expressions were consistently found until DCE-10 treatment. Therefore, it is presumed that DCE may have dynamic effects of innate immunity activation and pro-inflammation suppression on RAW264.7 cells simultaneously. These effects were consistently found in the highest dose of coffee, DCE-10 (equivalent to 10 cups of coffee a day in man), that might imply the small coffee molecules were accumulated in RAW 264.7 cells after DCE-10 treatment and produce synergistic cytokine effects for innate immunity activation and anti-inflammatory reaction concurrently.

Analysis of Marker Components of Fermented Opuntia ficus-indica var. saboten Stem Extracts (유산균 발효에 의한 손바닥선인장 줄기추출물의 지표물질 함량 변화 분석)

  • Shin, Dong Won;Lee, Sang Ho;Lee, Soyeon;Han, Eun Hye
    • Analytical Science and Technology
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    • v.31 no.6
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    • pp.219-224
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    • 2018
  • The fruit and stem of Opuntia ficus-indica var. aboten (OFS), a native plant of Jeju Island, are considered a safe food source. Moreover, stem extracts have been previously reported to possess a variety of biological effects (e.g. anti-inflammatory and anti-oxidant, including the ability to partially ameliorate cognitive impairment), suggesting that this plant may have utility as a functional food. The present study investigated whether fermentation by lactic acid bacteria enhances the biological effects of OFS extracts. The acetylcholinesterase (AChE) inhibitory activity of fermented or non-fermented OFS extracts was evaluated, and the content of marker components dihydrokaempferol (DHK) and quercetin-3-methyl ether (3-MeQ) was analyzed using high-performance liquid chromatography. Fermented (relative to non-fermented) OFS extracts exhibited improved AChE inhibitory activity ($IC_{50}=28.35 mg/mL$), with AChE inhibitory activity resulting from fermentation by L. plantarum ($IC_{50}=12.56mg/mL$) exceeding that resulting from fermentation by L. fermentum ($IC_{50}=17.71mg/mL$). Furthermore, fermented (relative to non-fermented) OFS extracts exhibited a 16.7 % increase in DHK content, and 3-MeQ content of OFS extracts fermented by L. plantarum and L. fermentum increased by 28.6 % and 21.4 %, respectively. Therefore, OFS stem extract AChE inhibitory activity, as well as DHK and 3-MeQ content, was enhanced by fermentation with Lactobacillus spp. This suggests that fermented OFS extracts may contribute to prevention or improvement of cognitive impairment. These data are anticipated to be useful in the development of enhanced-efficacy OFS products.

Arsenic species in husked and polished rice grains grown at the non-contaminated paddy soils in Korea (국내 비오염 논토양에서 재배한 현미와 백미 중 비소화학종 함량)

  • Kim, Da-Young;Kim, Ji-Young;Kim, Kye-Hoon;Kim, Kwon-Rae;Kim, Hyuck-Soo;Kim, Jeong-Gyu;Kim, Won-Il
    • Journal of Applied Biological Chemistry
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    • v.61 no.4
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    • pp.391-395
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    • 2018
  • There is an increasing concern over arsenic (As) contamination of paddy soil and rice with regard to food safety. This study was conducted to investigate total and inorganic As concentration in one hundred husked and polished rice samples collected at the non-contaminated paddy soil in Korea. Arsenic species in rice samples were extracted using 1% nitric acid ($HNO_3$) with a microwave oven and were measured using high performance liquid chromatography coupled with inductively coupled plasma-mass spectrometry. Mean concentrations of total As in husked rice and polished rice were 0.18 and $0.11mg\;kg^{-1}$, respectively. Also, average inorganic As concentrations in husked rice and polished rice were 0.11 and $0.07mg\;kg^{-1}$, respectively. These levels are lower than the standard guideline value 0.35 and $0.2mg\;kg^{-1}$ for inorganic As in husked and polished rice recommended by Codex Committee on Contaminants in Foods and Ministry of Food and Drug Safety, respectively. The mean of the inorganic As ratio for the total amount of As was 0.65 and 0.67 for husked rice and polished rice, respectively, and the range was from 0.08 to 1.0. For health risk assessment, the average value of cancer risk probability was $9.24{\times}10^{-5}$ and ranged from $2.30{\times}10^{-5}$ to $1.90{\times}10^{-5}$. Therefore, human exposure to As through dietary intake of surveyed rice samples might considered to be a low health risk.

The Effect of Fermented Extracts of Korean Dendropanax Morbifera Levéille on Hair Growth (황칠나무 발효 추출물의 육모효과)

  • Park, Tae-Hee;Park, Se-Ho;Lee, Jae-Yeul;Yang, Seun-Ah;Jhee, Kwang-Hwan
    • Journal of Life Science
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    • v.29 no.4
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    • pp.455-460
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    • 2019
  • In previous studies, we confirmed the effective antimicrobial activity of fermented Dendropanax morbifera leaf/branch extracts with Lactobacillus plantarum ilchiwhangchil 1785 and Lactobacillus plantarum ilchiwhangchil 2020. In this study, we investigated the hair growth effect of D. morbifera leaf/branch extracts fermented with L. plantarum ilchiwhangchil 1785 and L. plantarum ilchiwhangchil 2020 on human hair dermal papilla cells. The growth rate of human hair dermal papilla cells treated with fermented extracts in the range of 1 to $10{\mu}g/ml$ significantly increased in a concentration-dependent manner, without increasing cell death. Double staining studies showed that the growth of cells treated with fermented D. morbifera leaf/branch extracts was more active than that of control cells. Moreover, the cells treated with the fermented D. morbifera leaf/branch extracts exhibited a 18.84% and 23.31% increase in cell mobility, respectively, as compared with that of the untreated cells. High-performance liquid chromatography (HPLC) was used to determine the active agents responsible for hair growth. The results showed that the content of ${\beta}$-sitosterol, which is known to affect hair growth, increased about 10 times in the fermentation process of D. morbifera leaf/branch extracts. Taken together, the findings confirm that fermented Dendropanax morbifera leaf/branch extracts promote hair growth.

Effects of commonly used infusion method on catechin content and antioxidant capacities of pure green tea packaged in tea bags (음용 조건으로 추출한 티백 형태 순수 녹차의 카테킨 함량 및 산화방지능)

  • Kim, Mi-Seon;Rha, Chan-Su;Kim, Dae-Ok
    • Korean Journal of Food Science and Technology
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    • v.51 no.4
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    • pp.356-360
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    • 2019
  • To evaluate antioxidant capacities and total phenolic, flavonoid, and catechin contents of pure green tea packaged in tea bags, green tea bags of 10 commercial brands were purchased and analyzed. Considering commonly used infusion methods, green tea bags were extracted using 150 mL of spring water at $70^{\circ}C$ for 2 min. Ten brands of green tea showed a wide range of total phenolic content (46.3-93.9 mg gallic acid equivalents/g), total flavonoid content (10.3-21.7 mg catechin equivalents/g), and antioxidant capacities (88.8-175.6 mg and 87.9-183.5 mg vitamin C equivalents/g for DPPH and ABTS assays, respectively). Total catechin content of green tea ranged from 33.8-74.7 mg/g, which corresponded to 43.0-108.3 mg/150 mL of cup. Collectively, these results suggest that daily consumption of 3.8-12.5 cups of green tea packaged in tea bags meets the recommended daily intake of catechins in green tea extract as a health functional foods in Korea.

Anti-bacterial Activity and Standardization on Hwangryunhaedok-tang (황련해독탕의 항균활성 및 표준화 연구)

  • Lee, Dae-Yeon;Yi, Young-Woo;Kim, Wan-Su;Sim, Sun-Hyung;Kwon, Pil-Seung;Lee, In-Hee
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.2
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    • pp.214-220
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    • 2019
  • Hwangryunhaedok-tang (HT) is prescribed in oriental medicine to treat inflammation-related diseases. This study examined the anti-bacterial effect to select the index components of Traditional Chinese Medicine (TCM) HT. The disc diffusion method and minimum inhibition concentrations (MIC) tests were conducted to measure the antibacterial activity of HT. The strains were Staphylococcus aureus ATCC 25923 and Escherichia coli 23724. Berberine and geniposide were selected as the index components of HT and were analyzed quantitatively by high-performance liquid chromatography (HPLC). As a result, the inhibition ring was formed at $11.7{\pm}1.3mm$ and $8.7{\pm}0.7mm$ at HT 20 mg and 10 mg on S.aureus ATCC 25923, respectively; and $8.0{\pm}0.0mm$ at HT 20 mg on E.coli ATCC 23724. The MIC were 31.25 mg/mL in S.aureus ATCC 25923 and 125.0 mg/mL in E.coli ATCC 23724. In quantitative analysis for standardization, HT contained 16.55 mg/g of berberine and 81.85 mg/g of geniposide. Overall, when antibiotics are developed using natural products, they may reduce the side effects and inhibit resistant bacteria. If research on HT continuously proceeds, positive developments are expected to be made in oriental medicine as well as in various other industries.

Recombinant Production and Antimicrobial Activity of an Antimicrobial Model Peptide (Uu-ilys-CF) Derived from Spoon Worm Lysozyme, Uu-ilys (개불 라이소자임 유래 항균성 모델 펩타이드(Uu-ilys-CF)의 재조합 단백질 생산 및 항균 활성)

  • Oh, Hye Young;Go, Hye-Jin;Park, Nam Gyu
    • Journal of Life Science
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    • v.31 no.1
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    • pp.83-89
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    • 2021
  • Uu-ilys, an i-type lysozyme from spoon worm (Urechis unicinctus), is an innate immune factor that plays an important role in the defense against pathogens. It also possesses non-enzymatic antibacterial activity. Thus, there is a possibility to develop an antimicrobial model peptide from Uu-ilys. In this study, we report the design, production, and antibacterial activity of an Uu-ilys analog that exhibits antibacterial activity. The Uu-ilys structure was fragmented according to its secondary structures to predict the regions with antimicrobial activity using antimicrobial peptide (AMP) prediction tools from different AMP databases. A peptide containing the C-terminal fragment was predicted to exert antimicrobial activity. The chosen fragment was designated as an Uu-ilys analog containing the C-terminal fragment, Uu-ilys-CF. To examine the possibility of developing an AMP using the sequence of Uu-ilys-CF, recombinant fusion protein (TrxA-Uu-ilys-CF) was produced in an expression system that was heterologous. The produced fusion protein was cleaved after methionine leaving Uu-ilys-CF free from the fusion protein. This was then isolated through high performance liquid chromatography and reverse phase column, CapCell-Pak C18. The antibacterial activity of Uu-ilys-CF against different microbial strains (four gram-positive, six gram-negative, and one fungal strain) were assessed through the ultrasensitive radial diffusion assay (URDA). Among the bacterial strains tested, Salmonella enterica was the most susceptible. While the fungal strain tested was not susceptible to Uu-ilys-CF, broad spectrum antibacterial activity was observed.

Protective effects of extracts from the aerial parts of hydroponically cultured ginseng on alcohol-induced liver damage in mice and quantitative analysis of major ginsenosides (알코올성 간손상을 유발한 마우스 모델에서의 새싹인삼 지상부 추출물의 간 기능 보호효과 및 지표성분 함량분석)

  • Lee, Mi Kyoung;Jang, In-Bae;Lee, Min Ho;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.63 no.4
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    • pp.413-420
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    • 2020
  • We studied the effects of the extract of aerial parts of hydroponically cultured ginseng (HGE) on alcohol-induced liver damage (AILD) in mice. AILD was induced by the oral administration of ethanol (EtOH) (25%; 5 g/kg body weight) for seven days in the study as well as EtOH-only groups. However, HGE (4 and 12 mg/kg) was orally administered (once daily for ten consecutive days) only to the study group, three days prior to the EtOH treatment. The HGE-treated group showed significantly lower levels of alanine aminotransferase and aspartate aminotransferase than the EtOH-only group. In addition, HGE administration decreased the level of serum lactate dehydrogenase, a known marker of liver damage. The effect of HGE on AILD was found to be dose dependent, and the consecutive administration of HGE showed no side effects in mice. Our study indicates that HGE treatment can potentially reduce oxidative stress and toxicity in the liver of alcohol-treated mice and that HGE can be a useful therapeutic agent for alcohol-induced hepatotoxicity. Additionally, a simple and efficient high-performance liquid chromatography-ultraviolet detection method was developed for determining the contents of four major ginsenosides in HGE. The aerial parts of hydroponically cultured ginseng were extracted using 70% fermented ethanol, and the contents of ginsenosides F5, F3, F1, and F2 in HGE were found to be 2.5, 4.4, 1.4, and 23.3 mg/g, respectively.