• Title/Summary/Keyword: HPLC Chromatography

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Protective effect of ethyl acetate fraction from Actinidia arguta sprout against high glucose-induced in vitro neurotoxicity (포도당으로 유도된 in vitro 뇌신경세포 독성에 대한 다래 순 아세트산에틸 분획물의 보호 효과)

  • Yoo, Seul Ki;Park, Seon Kyeong;Kim, Jong Min;Kang, Jin Yong;Park, Su Bin;Han, Hye Ju;Kim, Chul-Wo;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.50 no.5
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    • pp.517-527
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    • 2018
  • The current study investigated in vitro anti-diabetic and neuroprotective effects of the ethyl acetate fraction in Actinidia arguta sprouts (EFAS), on $H_2O_2$ and high glucose-induced cytotoxicity in human neuroblastoma MC-IXC cells. EFAS had high total phenolic and total flavonoid contents. An assessment of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity of EFAS, as well as its potential for inhibiting malondialdehyde production, indicated that EFAS may possess significant antioxidant properties. EFAS exerted inhibitory effects on ${\alpha}-glucosidase$ via glycemic regulation which forms advanced glycation end products. In addition, EFAS exhibited significant acetylcholinesterase inhibitory effects. Moreover, EFAS displayed protective effects against $H_2O_2$ and high glucose-induced cell death, and inhibited the generation of reactive oxygen species in MC-IXC cells. Finally, the main physiological compound of EFAS was identified via high performance liquid chromatography as a rutin.

Postprandial hypoglycemic effects of mulberry twig and root bark in vivo and in vitro (In vivo와 in vitro에서 상지 및 상백피 에탄올추출물이 식후 혈당 상승 억제 조절에 미치는 영향)

  • Park, Soo Yeon;Jin, Bo Ra;Lee, Yu Rim;Kim, You Jin;Park, Jeong Bin;Jeon, Young Hee;Choi, Sang Won;Kwon, Oran
    • Journal of Nutrition and Health
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    • v.49 no.1
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    • pp.18-27
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    • 2016
  • Purpose: Our previous study demonstrated the hypoglycemic effects of mulberry (Morus alba L.) leaf and the underlying mechanisms. Here we explored the potency of mulberry twigs (TW) and root barks (RB) in postprandial hypoglycemic effects in vitro and in vivo. Methods: The major components of TW and RB were determined by high performance liquid chromatography (HPLC). Alpha-glucosidase inhibition and glucose/fructose uptake inhibition in Caco-2 cells were determined for TW, RB, and their major components, followed by an oral sugar tolerance test (OSTT) in streptozotocin-induced diabetic rats. Male Wistar rats were fed a high-fat diet for 2 weeks and then a single dose of streptozotocin (35 mg/kg B.W) was administered by intraperitoneal injection. Rats with fasting blood glucose levels above 126 mg/dL were randomly divided into 5 groups (n = 8/group) for the following treatments by gavage for 4 weeks: vehicle (normal control and diabetic control), 200 mg/kg B.W of TW or RB or 100 mg/kg B.W of oxyresveratrol (OXY). Results: OXY and mulberroside A were identified as the major components of TW and OXY, mongolicin, and kuwanon H for RB. A significant inhibitory activity on ${\alpha}-glucosidase$ was found for TW, RB, and OXY (p = 0.0099). There was a dose-dependent inhibition of TW and RB on the intestinal sugar uptakes in Caco-2 cells, showing a greater impact on fructose compared to glucose. The OSTT showed that TW and RB significantly delayed time to maximal concentration (p = 0.0088) and decreased maximal concentration (p = 0.0043) compared to the control group. Conclusion: These results suggest that TW and RB may have a postprandial hypoglycemic effect, particularly in the case of high fructose or sucrose intake. OXY was suggested as a contributor to the hypoglycemic effect of TW and RB. Further studies are needed for the systemic effect of TW and RB in circulation.

Cell Protective Effects of Enzymatic Hydrolysates of Citrus Peel Pectin (귤피 펙틴 유래 효소적 가수분해물의 세포 보호 효과)

  • Kwon, Soon Woo;Ko, Hyun Ju;Bae, Jun Tae;Kim, Jin Hwa;Lee, Geun Soo;Pyo, Hyeong Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.1
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    • pp.75-85
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    • 2016
  • Pectin, a naturally occurring polysaccharide, has in recent years attracted considerable attention. Its benefits are increasingly appreciated by scientists and consumers due to its safety and usefulness. The chemistry and gel-forming characteristics of pectin have enabled to be used in pharmaceutical industry, health promotion and treatment. Yet, it has been rarely used in cosmetics because of its incompatibility with many cosmetic ingredients, including alcohols, and unstable viscosity of pectin gels under various pH and salt conditions. However, low-molecular-weight pectin oligomers have excellent biological activities, and depolymerization of pectin to produce cosmetic ingredients would be very useful. In this study, we attempted the development of cosmetic ingredients using pectin with an excellent effect on human skin. We developed a bio-conversion process that uses enzymatic hydrolysis to produce pectin hydrolysates containing mainly low-molecular-weight pectin oligomers. Gel permeation chromatography was used to determined the ratio of hydrolysis. The molecular weight of the pectin hydrolysates obtained varied between 200 and 2,700 Da. The two newly developed low-molecular-weight pectin hydrolysates, LMPH A and B, had higher anti-oxidative activities than pectin or D-galacturonic. Exposure to UVB radiation induces apoptotic cell death in epidermal cells. Annexin V binding and propidium iodide uptake were measured by flow cytometry to evaluate UVB-induced cell death in HaCaT cells. Both LMPH A and B reduced UVB-induced cell death and increased cell proliferation by 22% and 30% at 0.5% concentration respectively, while pectin had no significant activity. In conclusion, this study suggests that the newly developed low-molecular-weight pectin hydrolysates can be used as safe and biologically active cosmetic ingredients.

Neuronal Cell Protective Effect of Dendropanax morbifera Extract against High Glucose-Induced Oxidative Stress (High Glucose로 유도된 산화 스트레스에 대한 황칠나무 잎 추출물의 뇌신경세포 보호 효과)

  • Kim, Jong Min;Park, Seon Kyeong;Guo, Tian Jiao;Kang, Jin Yong;Ha, Jeong Su;Lee, Du Sang;Kwon, O-Jun;Lee, Uk;Heo, Ho Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.7
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    • pp.938-947
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    • 2016
  • Antioxidant activities and neuroprotective effects of ethyl acetate fraction from Dendropanax morbifera (EFDM) against high glucose-induced oxidative stress and neurotoxicity were investigated to confirm their physiological activities. An 80% ethanolic extract of D. morbifera showed the highest contents of total phenolic compounds as well as 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 1,1-diphenyl-2-picrylhydrazyl radical scavenging activities. The extract was fractionated using several solvents, and the ethyl acetate fraction showed the highest activities in ferric reducing/antioxidant power and malondialdehyde inhibitory assays. To evaluate the neuroprotective effect based on antioxidant activities, cell viability was assessed using PC12 and MC-IXC cells in $H_2O_2$- and high glucose-induced cytotoxic assays, respectively. EFDM evidently showed neuroprotective effects in all cells (neuron-like PC12 cells and human brain-originated neuroblastoma MC-IXC cells). Inhibitory effect of the extract on acetylcholinesterase (AChE) as an acetylcholine-hydrolyzing enzyme was performed to examine the effect on cognitive function. EFDM presented an AChE inhibitory effect. Finally, high-performance liquid chromatography analysis showed that the major phenolic compound of EFDM is probably a rutin.

Effects of Extraction Method on Anserine, Protein, and Iron Contents of Salmon (Oncorhynchus keta) Extracts (연어(Oncorhynchus keta) 추출물 중의 Anserine, 단백질 및 철분 함량에 미치는 추출방법의 영향)

  • Min, Hye-Ok;Park, In-Myoung;Song, Ho-Su
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.2
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    • pp.220-228
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    • 2017
  • Effects of extraction methods on reducing concentrations of pro-oxidants (total iron and protein) of salmon was determined. For development of the extraction process, the effectiveness of several extraction methods was determined and compared, including heat treatment (60, 80, and $100^{\circ}C$), ion exchange and carboxymethyl (CM)-cellulose column chromatography, and ultrafiltration (UF). Protein, total iron, and anserine contents of salmon extracts were 23.64 mg/mL, $16.20{\mu}g/mL$, and 5.47 mg/mL in non-heated extracts, 7.40 mg/mL, $2.32{\mu}g/mL$, and 5.20 mg/mL in heated extracts at $60^{\circ}C$, 7.64 mg/mL, $1.20{\mu}g/mL$, and 5.21 mg/mL at $80^{\circ}C$, and 7.04 mg/mL, $0.68{\mu}g/mL$, and 4.04 mg/mL at $100^{\circ}C$, respectively. Heating and UF decreased contents of protein and total iron, whereas only UF slightly decreased anserine content. Application of the primary ion exchange method increased the content of anserine up to 16%. Protein and total iron contents by the primary ion exchange method decreased by 70 and 98%, respectively. Secondary ion exchange (CM-cellulose) treatment after primary ion exchange and UF resulted in lower anserine content than the primary ion exchange method. However, the content of impurities (protein, total iron) was lower than in all other salmon extracts. Therefore, primary ion exchange, UF, and secondary ion exchange method were the best extraction processes in this study.

Thiobarbituric Acid Reactive Substances Levels in Brain Tissue of Aldh2 Knockout Mice Following Ethanol Exposure for 8 Weeks (Aldh2 knockout 마우스에서 8주간 에탄올 노출에 따른 뇌조직의 thiobarbituric acid reactive substances 농도)

  • Moon, Sun-In;Eom, Sang-Yong;Kim, Jung-Hyun;Yim, Dong-Hyuk;Kim, Hyong-Kyu;Kim, Yong-Dae;Kim, Heon
    • Journal of Life Science
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    • v.21 no.8
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    • pp.1163-1167
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    • 2011
  • Excessive alcohol consumption causes various degenerative brain diseases including Alzheimer's disease and Parkinson's disease. Absorbed ethanol is metabolized to acetaldehyde and acetic acid by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). Acetaldehyde is well known as a toxicant through generation of reactive oxygen species (ROS). Therefore, ALDH2 activity may play important roles in the pathogenesis of alcohol-induced brain diseases. In this study, we demonstrated the effects of ALDH2 enzyme activity on lipid peroxidation in brain tissues and urine of mice exposed to ethanol for 8 weeks. Five male, 8-week old Aldh2 (+/+) and Aldh2 (-/-) mice (C57BL/6J strain) in each group were exposed to ethanol for 8 weeks (2 g/kg wt./day) using gavage, and those in the control group received 0.9% saline alone. Thiobarbituric acid reactive substances (TBARS) level, a marker for lipid peroxidation, was measured in whole brain tissue and urine by high performance liquid chromatography. As a result, chronic ethanol treatment did not show any statistical change on the TBARS level of brain tissue in both Aldh2 (+/+) mice and in Aldh2 (-/-) mice. However, following ethanol exposure for 8 weeks in Aldh2 (-/-) mice, the urinary TBARS levels were significantly increased to more than double compared to the pretreatment group. This result was not observed in Aldh2 (+/+) mice. These results suggest that although ALDH2 enzyme activity plays a role in the generation of ROS in the whole body, it does not seem to be important in the pathogenesis of alcohol induced degenerative brain diseases.

Method Development for the Sample Preparation and Quantitative Analysis of Synthetic Colors in Foods (식품 중 식용타르색소의 시료별 전처리방법 확립 및 함량 분석)

  • Park, Sung-Kwan;Lee, Tal-Su;Park, Seung-Kook
    • Korean Journal of Food Science and Technology
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    • v.36 no.6
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    • pp.893-899
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    • 2004
  • Method for sample preparation and quantitative analysis of 19 permitted and non-permitted synthetic colors in foods was developed based on reversed-phase ion-pairing high performance liquid chromatography. For color extraction of samples, deionized water was added, and pH was appropriately adjusted with 1% ammonia water. Any undissolved matters were extracted with 50% ethanol or 70% methanol. Lipid in snacks was first removed using n-hexane with centrifugation, water was added to extract colors, followed by clean-up and concentration using Sep-Pak $C_{18}$ cartridge. Recovery efficiencies at known concentrations of 19 standard food colors spiked into foods were in 90.3-97.9% range far soft drink, 79.2-101.9% for candy, 84.1-103.4% for jelly, 86.4-100.8% for chewing gum, 83.5-103.4% for ice cream, and 78.5-95.6% for snack.

Quality Changes in Red Ginseng Extract during High Temperature Storage (열처리(熱處理)에 의한 홍삼(紅蔘)엑기스의 성분변화(成分變化))

  • Choi, Jin-Ho;Kim, Woo-Jung;Yang, Jae-Won;Sung, Hyun-Soon;Hong, Soon-Keun
    • Applied Biological Chemistry
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    • v.24 no.1
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    • pp.50-58
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    • 1981
  • The influence of high temperature storage on the chemical composition and color intensity of the concentrated red ginseng extract(RGE) was investigated. The concentrated RGE was prepared by extraction of red ginseng tails with water and concentrated under reduced pressure. Changes in free sugars, saponin patterns and brown color intensity were measured during 96 hours of heat treatment at various temperature. A decrease in the contents of glucose, fructose and sucrose was resulted as the brown color intensity increased during the storage. The sugar contents and color intensity showed rapid initial change followed by slowing down at higher temperature. A significant relationship was found between sugar content and browning rate. The saponin pattern measured by high performance liquid chromatography, particularly in the region of protopanaxtriol, was also affected significantly. The peak heights of ginsenoside -Re and $-Rg_1$ were decreased while those of ginsenoside $-Rg_2$ and -Rh group were increased.

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Antioxidants in Serum and Induced Sputum of COPD Patients (만성폐쇄성폐질환 환자의 혈청 및 유도객담에서의 항산화제)

  • Park, Hyeon-Kwan;Yu, Young-Kwon;Kim, Kyu-Sik;Lim, Sung-Chul;Kim, Young-Chul;Park, Kyung-Ok
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.2
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    • pp.158-170
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    • 2001
  • Background : Although an oxidants and antioxidants imbalance has been considered in the pathogenesis of chronic obstructive pulmonary disease (COPD), there is a paucity of reports focussing on the smoking-induced changes of oxidants and antioxidants in COPD. Method : The concentration of antioxidants (ascorbic acid, uric acid, retinol, and $\alpha$- & $\gamma$-tocopherol) was measured in the serum and induced sputum of 30 healthy controls and 34 stable COPD patients using high performance liquid chromatography (HPLC). The inhibition of lipid peroxidation as an index of antioxidant capacity was measured in the serum by a TBA assay. Results : The serum concentration of ascorbic acid, $\alpha$-tocopherol, and retinol were significantly lower in the patients with COPD than in healthy controls ($484.8{\pm}473.3$ vs $1497.8{\pm}819.2\;pmol/L$, $48.38{\pm}17.34$ vs $73.96{\pm}26.29\;pmol/L$, p<0.001, and $9.51{\pm}8.33$ vs $15.01{\pm}5.88\;pmol/L$, p<0.05, respectively, mean$\pm$SD). However, there were little differences in the ascorbic acid and uric acid concentrations in the induced sputum between the COPD patients and the controls. The induced sputum to serum ratio of ascorbic acid was significantly higher in COPD patients compared with healthy control (0.375 vs 0.085, p<0.05). In the normal controls, the serum ascorbic acid concentration was lower in smokers than in nonsmokers ($1073{\pm}536$ vs $1757{\pm}845\;pmol/L$, p<0.05), but the level was still higher than that of the COPD patients (p<0.05). The serum retinol levels were correlated with $FEV_1$ in COPD patients (r=0.58, p<0.05). The products of lipid peroxidation were increased in normal smokers and COPD compared with norma1 nonsmokers ($115.56{\pm}19.93$ vs $120.02{\pm}24.56$ vs $91.87{\pm}20.71\;{\mu}mol/{\mu}mol$ Pi of liposome, p<0.05). Conclusion : Cigarette smoking may induce the dep1etion of serum antioxidants and this depletion of antioxidants is suggested to play a role in the pathogenesis of COPD.

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Skin Whitening Effect of Ethyl Acetate Fraction of Adenophora triphylla var. japonica Sprout (잔대(Adenophora triphylla var. japonica)순 아세트산에틸 분획물의 피부 미백 효과)

  • Yoo, Seul Ki;Park, Seon Kyeong;Kang, Jin Yong;Kim, Jong Min;Park, Sang Hyun;Kwon, Bong Seok;Lee, Chang Jun;Kang, Jeong Eun;Park, Su Bin;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Plant Resources
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    • v.30 no.4
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    • pp.352-363
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    • 2017
  • To investigate skin-whitening effect of Adenophora triphylla var. japonica sprout extract, antioxidant activity, inhibitory effect on tyrosinase and melanin synthesis in B16/F10 melanoma cell were examined. Total phenolic content (246.25 mg GAE/g) and total flavonoid content (303.94 mg RE/g) of ethyl acetate fraction from Adenophora triphylla sprout (EFAT) showed the highest contents than other fractions (n-hexane, chloroform and distilled water). Antioxidant activities of EFAT has been evaluated using ABTS, DPPH radical scavenging activities, FRAP and inhibitory effect of lipid peroxidation. EFAT showed excellent radical scavenging activity and inhibitory effect on MDA production. Inhibitory effect of tyrosinase as a major enzyme of melanin synthesis was also measured. In these results, EFAT showed higher inhibitory effect against L-DOPA (51.27%) than L-tyrosine. $IC_{50}$ value on ${\alpha}-glucosidase$ was $41.93{\mu}g/ml$. In B16/F10 melanoma cells, EFAT inhibited melanin synthesis at $200{\mu}g/ml$ concentration (about 42% decrease). Finally, main physiological compounds of EFAT were identified as a rutin and a chlorogenic acid using high performance liquid chromatography.