• Title/Summary/Keyword: chromatography test

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Effect of Isoquercitrin-containing Nelumbo nucifera Leaves Extract on Skin Wrinkle Improvement (Isoquercitrin 함유 연꽃잎 추출물의 피부주름개선 효능 연구)

  • Moon, Eunjung;Jeon, Junmyeong;Lee, Gahee;Baik, Minyoung;Lee, Dae Woo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.2
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    • pp.191-200
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    • 2018
  • In this study, we conducted research to find anti-wrinkle skin care ingredients from natural sources via the measurement of procollagen type 1 levels in the medium of normal human dermal fibroblast (NHDF) and found that a Nelumbo nucifera leave extract (NLE) was as the best effective ingredient. By the high performance liquid chromatography analysis, we confirmed isoquercitrin was one of the main compounds of NLE. Moreover, it also increased the procollagen type 1 production without cytotoxicity of NHDF. NLE and isoquercitrin exerted free radical scavenging activity. Especially, isoquercitrin exhibited strong intracellular antioxidant capacity in human skin-derived cells, HaCaT and NHDF. Finally, we performed clinical test for the inhibitory effect of NLE on skin wrinkle formation. A randomized study was conducted in 22 healthy female volunteers, aged between 30 and 65 yrs, with moderate to moderately severe facial wrinkles. Volunteer applied a 1.7% NLE cream (isoquercitrin 51 ppm) and a control cream at each facial side (left/right) twice daily for 8 weeks. The 1.7% NLE cream improved skin roughness through reducing the wrinkle of the craw's feet significantly without any skin side effect. Our results demonstrate that NLE and isoquercitrin can increase the collagen production and exert antioxidant activity. Therefore, we expect that the new non-toxic herbal extract, isoquercitrin-containing NLE will be interesting natural ingredient of cosmetics with anti-wrinkle efficacy.

Enzymatic Synthesis and Characterization of Structured Lipids from Docosahexaenoic Acid - Enriched Fish oil and Soybean oil (어유와 대두유로부터 기능성 유지의 효소적 합성)

  • Kim, Yu-Mi;Jeon, Mi-Sun;Lee, Jeung-Hee;Lee, Ki-Teak
    • Food Science and Preservation
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    • v.15 no.3
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    • pp.437-444
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    • 2008
  • Structured lipids(SLs) were synthesized by enzymatic interesterification with DHA-enriched fish oil(containing 27% docosahexaenoic acid) and soybean oil in the hatch-type reactor. The interesterification was performed for 24 hr at $55^{\circ}C$ and TLIM(immobilized lipase from Thermonyces lanuginosa, 10% by weight of total substrates) was mixed with 180 rpm of shaking. The fish oil and soybean oil were interesterifed with several weight ratio(fish oil : soybean oil, 2:8, 3:7, 4:6, 5:5, w:w), Reverse-phase high performance liquid chromatography with an evaporative light-scattering detector separated the triglyceride species of SLs. The products contained the newly synthesized peaks. Especially, one of peaks was distinctively increased with the increasing weight ratio from 2:8 to 5:5 while the peak of trilinolein (LLL) decreased vice versa. The effect of antioxidants such as catechin, BHT(Butylated hydroxytoluene), and their combinations on the oxidative stability in SL were investigated. Oxidative stability was carried out under oven test at $60^{\circ}C$ over 72 hr thereafter SLs were analyzed for total fatty acid content, rancimat, peroxide value, electronic nose and TBARS value. Among all combinations of antioxidant, the highest stability was obtained from 200 ppm of catechin. Besides, total tocopherol ($\alpha$, $\gamma$, and $\delta$-tocopherol), iodine and saponification value were analyzed in which iodine and saponification value of SLs were 151.19 and 182.35.

Inhibitory Effects of Carex humilis Extract on Elastase Activity and Matrix Metalloproteinase-1 Expression (산거울 추출물의 Elastase 활성 저해 및 Matrix Metalloproteinase-1 발현 억제 효과)

  • Park, Sun-Hee;Lee, Kang-Hyuk;Han, Chang-Sung;Kim, Ki-Ho;Kim, Young-Heui
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.2
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    • pp.129-136
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    • 2010
  • In order to evaluate anti-wrinkle activity of Carex humilis extract, free radical scavenging activity, elastase inhibitory activity and reduction of expression Matrix metalloproteinase-1 (MMP-1) mRNA and MMP-1 protein were investigated. The roots of Carex humilis were extracted with 95 % ethanol and successively partitioned with organic solvents with increasing polarity of the solvents. Each fraction of organic solvent were investigated by using free radical scavenging activity and elastase inhibitory activity test. Among them, EtOAc fraction showed antioxidant activity ($SC_{50}$=4.89 ${\mu}g/mL$) and elastase inhibitory activity ($IC_{50}$=23.5 ${\mu}g/mL$). EtOAc fraction was developed on silica gel by open-column chromatography and consecutively re-developed on C18 resin by prep-HPLC to give ${\alpha}$-viniferin as a major component, which was confirmed by spectrometric analysis. In the assay on expression of MMP-1 mRNA by RT-PCR and protein by western-blot, EtOAc layer (10 ~ 100 ${\mu}g/mL$) was reduced about 50 ~ 60 %, 50 ~ 65 % respectively and ${\alpha}$-viniferin (0.5 ~ 2 ${\mu}g/mL$) was inhibited about 60 ~ 75 %, 55 ~ 65 % respectively in human fibroblast. Therefore, our findings suggest that EtOAc layer of Carex humilis containing ${\alpha}$-viniferin can be useful as an active ingredient for cosmeceuticals of anti-wrinkle effects.

Concentration of Polycyclic Aromatic Hydrocarbons in Cereals, Pulses, Potatoes, and their Products (곡류, 두류, 서류 및 그 가공품 중 다환 방향족 탄화수소 (polycyclic aromatic hydrocarbons) 화합물 함량)

  • Kim, Hee-Yun;Chung, So-Young;Sho, Yo-Sub;Park, Seong-Soo;Lee, Eun-Ju;Suh, Jung-Hyuk;Lee, Yun-Dong;Choi, Woo-Jeong;Kim, Jung-Soo;Eom, Ji-Yoon;Park, Hui-Og;Jin, Myeong-Sig;Kim, Dong-Sul;Ha, Sang-Chul;Lee, Jong-Ok
    • Korean Journal of Food Science and Technology
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    • v.37 no.4
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    • pp.537-541
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    • 2005
  • Estimation of concentrations of PAHs [benzo(a)anthracene, chrysene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(a)pyrene, dibenzo(a,h)anthracene, benzo(g,h,i)perylene, indeno(1,2,3-c,d)pyrene] in cereals, pulses, potatoes, and their products available in Korean markets gave mean levels of 0.13, 0.08, 0.06, 0.03, 0.08, 0.15, 0.45, and $0.14{\mu}g/kg$, respectively, with recoveries between 82.6-106.6%. Methodology involved saponification and extraction with n-hexane, purification on Sep-Pak Florisil cartridges, and high performance liquid chromatography using a fluorescence detector.

Reduction of Antigenicity of Bovine Casein by Microbial Enzymes (미생물효소에 의한 우유 casein의 항원성 저감화)

  • Choe, Hyeon-Seok;Ahn, Jong-Nam;Jeong, Seok-Geun;Ham, Jun-Sang;In, Yeong-Min;Kim, Dong-Un
    • Journal of Dairy Science and Biotechnology
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    • v.21 no.2
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    • pp.97-104
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    • 2003
  • It is extremely important to destroy the antigenicity of milk proteins for dietetic treatment of infants with milk allergy. Enzymatic digestion of milk protein is not only effective for destroying antigenicity, but it also is less liable to alter the nutritive value. Bovine casein was hydrolyzed with eight different commercial proteases derived from bacterias or fungi, either individually or in combination to eliminate protein allergenicity. The average molecular weight of casein hyrdolysates determined by size exclusion chromatography is about 550${\sim}$2,300 dalton range. Antigenicity of the casein hyrdolysates was not detected by heterologous passive cutaneous anaphylaxis in guinea pig-rabbit antiserum system. The inhibition test on the enzyme-linked immunosorbent assay(ELISA) showed that the antigenicity of casein hydrolysates is lowed up to 1/8,000 than that of intact bovine casein. As the enzyme reaction was carried out by the combination of bacterial and fungal protease, casein hydrolysates showed much lower bitterness and antigenicity. It suggests that these hydrolysates will be applied to many kinds of foods including the development of hypo-allergenic infant formula.

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The Allelopathic Effect of Alfalfa residues on Crops and Weeds (잡초와 작물에 대한 알팔파 잔유물의 Allelopathy 효과)

  • Yu, C.Y.;Jeon, I.S.;Chung, I.M.;Hur, J.H.;Kim, E.H.
    • Korean Journal of Weed Science
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    • v.15 no.2
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    • pp.131-140
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    • 1995
  • This experiment was conducted to test the effect on germination and seedling growth of major industrial crops and weed control potential using alfalfa plant. When dried alfalfa residues were mixed into vermiculite, germination, length of shoot and root of crops, such as Perilla frutescens, Sorghum nevosum, Platycodon grandiflorum and weeds, Digitaria saguinalis, Setaria viridis, Siegesbechia pubescens, Ammaranthus lividus, and Solanum nigrum, were significantly inhibited as the dried residue concentration increased. More than 10% concentration of the dired residue caused 80% germination and growth inhibition. The fresh alfalfa exudation also inhibited the germination and seedling growth of crop, barley, rye, alfalfa, and sesame, and weeds, Echinochloa crus-galli, Siegesbechia viridis, and Portulaca oleracea. The degree of inhibition showed the different response according to the fresh exudation concentration, types of crops and weeds. Generally, as the exudation concentration increased, the germination and seedling growth of crops and weeds inhibited. The exudation of dried residue also exhibited the strong inhibition effect on germination and seedling growth of crops, alfalfa, Platycodon grandiflorum, barley, sesame, rye and weeds, D. sanguinalis, S. pubescens, S. viridis, P. oleracea, E. crus-galli, At the 10% concentrations, S. pubescens, and P. oleracea were not germinated and showed only 15% germination in the S. viridis. From this study, would conclude that alfalfa plant contained water soluble phytotoxic substances which were inhibitory to weeds and crops. This results suggest that alfalfa had some possibility to control some weed species using toxic compounds like natural herbicide.

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Simultaneous determination and acute toxicity study of Fructus mume extracts in ICR mice (오매(烏梅)의 다성분동시분석 및 마우스를 이용한 경구 단회투여 급성독성시험)

  • Lee, In-Sun;Han, Chang-Hyun;Lee, Chul;Hwang, Bang-Yeon;Jung, Sang-Hyeok;Lee, Young-Joon;Jeon, Won-Kyung
    • Journal of Society of Preventive Korean Medicine
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    • v.15 no.1
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    • pp.37-47
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    • 2011
  • Objectives : Previous studies have shown that Fructus mume (FM) has anti-platelet effects. The present study was performed to determine the acute oral toxicity and quality control of a crude extract of FM in ICR mice. Methods : We investigated the in vivo single dose acute toxicity of FM 95% ethanol extract. This test was orally administered once by gavage to 20 male and 20 female mice at dose levels of 0 (control group), 1250, 2500 and 5000mg/kg body weight, respectively. Mortalities, clinical findings, autopsy findings and body weight changes were monitored daily for the 14 days following the administration. HPLC analysis was performed for the simultaneous determination of ursolic acid and p-hydroxycinnamic acid in FM. Reverse-phase chromatography using a C18 column and photodiode array detection at 211 nm was used for quantification of the two maker components. The mobile phase for gradient elution consists of water and acetonitrile. Results : We observed survival rates, general toxicity, change of body weight, and autopsy. The mice did not die after single oral administration of maximum dose of FM. Autopsy of animal revealed no abnormal gross finding. Therefore, $LD_{50}$ value of FM for ICR mice was more than 5000mg/kg on oral route. The HPLC analysis showed that ursolic acid and p-hydroxycinnamic acid amounts to 9.75- and 0.12% in the extract with the retention times of 47.99- and 15.38 minutes, respectively. Conclusions : These results suggest that no toxic dose level of FM in mice is considered to be more than 5000mg/kg. Consequently, it was concluded that FM have no effect on acute toxicity and side effect in ICR mice. For the quality control of FM extract, simultaneous determination of ursolic acid and p-hydroxycinnamic acid was established.

Determination of Free 4-hydroxyproline with Dansylchloride by HPLC in Human Urine (소변 중 4-hydroxyproline 분석에 관한 연구)

  • Lee, Keou-Weon;Cho, Young-Bong;Lee, Kyung-Jong
    • Journal of Preventive Medicine and Public Health
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    • v.35 no.4
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    • pp.282-286
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    • 2002
  • Objectives : The level of 4-hydroxyproline (4-Hyp) in human urine was measured using high performance liquid chromatography (HPLC) with a fluorescence detector. This method is useful for medical examinations and investigating the radicals induced by physical, chemical, mental stresses. This method is superior to many published several methods in terms of its low cost and ability to analyze many samples. Methods : The urine from workers in a tire manufacturing company (22 male pre- and post-shift workers) and 18 office-workers as controls were analyzed. Data concerning age, the cumulative drinking amount and the cumulative smoking amount was collected with a questionnaire. The optimum applied amount of dansyl-Cl, the optimum reaction temperature and time, the recoveries and the optimum pH of the eluent and buffer were determined.4-Hyp from human urine was derivatized with dansyl-Cl (dimethylamino-naphthalene-1-sulfonyl chloride) after removing the a-amino acid by a treatment with phthalic dicarboxaldehyde (OPA) and cleaned with Bond Elut C18 column. The 4-Hyp derivatives were separated on a reversed phase column by gradient elution with a phosphate buffer (5 mmol, pH 8.0) and acetonitrile, and detected by fluorescence measurements at 340 nm (excitation) and 538 nm (emission). Results : The detection limit for the urinary free 4-Hyp was $0.364{\mu}mol/l$. The recovery rate of 4-Hyp was 99.7%, and the effective pH of the phosphate buffer and borate buffer were 3.0 and 8.0, respectively. From statistical analysis, age, drinking and smoking did not affect the urinary free 4-Hyp in both the controls and workers. The range of urinary 4-Hyp in the controls, pre-shift, and post-shift workers were 0.33-16.44, N.D-49.06, and $0.32-56.27{\mu}mol/l$. From the pared-sample t-test, the urinary 4-Hyp levels in post-shift workers ($11.82{\pm}6.73\;nmmol/mg\;Cre$) were 2-fold higher than in pre-shift workers ($5.36{\pm}5.53\;nmol;/mg\;Cre$) and controls ($4.91{\pm}4.89\;nmol;/mg\;Cre$). Conclusions : This method was developed with high sensitivity, accuracy, and precision. The present method was effectively applied to analyze the urinary free 4-Hyp in both controls and workers.

Optimization of Onion Oil Microencapsulation by Response Surface Methodology (반응표면분석법에 의한 양파유 미세캡슐화 공정의 최적화)

  • Hong, Eun-Mi;Yu, Mun-Gun;Noh, Bong-Soo;Chang, Pahn-Shick
    • Korean Journal of Food Science and Technology
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    • v.34 no.3
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    • pp.437-443
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    • 2002
  • Using agar and gelatin as wall materials, onion oil was microencapsulated using the extrusion spraying technology. A sensitive methodology was developed for quantitative determination of the microencapsulation yield through ethyl acetate extraction and gas chromatographic analyses. Optimal conditions for the microencapsulation process consisting of the ratio of [core material, Cm] to [wall material, Wm] ($X_1$), temperature of dispersion fluid ($X_2$), detergent concentration in dispersion fluid ($X_3$), and concentration of emulsifier $(X_4)$ were determined using response surface methodology. The regression model equation for the yield of microencapsulation (Y, %) of onion oil could be predicted as $Y\;=\;97.028571-0.775000\;(X_1)-0.746726\;(X_1){\cdot}(X_1)\;-\;1.100000\;(X_3){\cdot}(X_2)$. The optimal conditions for the microencapsulation of the onion oil were determined as the ratio of [core material] to [wall material] of 4.5 : 5.5 (w/w), the temperature of dispersion fluid of $17.1^{\circ}C$ detergent concentration in dispersion fluid of 0.03%, and the concentration of emulsifier of 0.42%. Results revealed the most stable microcapsule of onion oil could be formed with the highest yield of microencapsulation (more than 95%) under optimal conditions.

Elucidation of Anti-tumor Initiator and Promoter Derived from Seaweed-2: Investigation of Seaweed Extracts Suppressing Mutagenic Activity of PhIP and MeIQx (해조류 중의 anti-tumor initiator 및 promoter의 해석-2: PhIP와 MeIQx의 돌연변이원성을 억제하는 해조 추출물 소재의 연구)

  • PARK Young-Beom;KIM In-Soo;YOO Sung-Jae;AHN Jong-Khan;LEE Tae-Gee;PARK Douck-Chon;KIM Seon-Bong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.4
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    • pp.581-586
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    • 1998
  • The present study was conducted to prepare seaweed extracts suppressing mutagenic activity of 2-amino-1-methyl-6-phenylimidazo[4,5-b] pyridine ( PhIP ) ana 2-amino-3,8-dimethylmidazo[4,5-f]quinoxaline (MeIQx) derived from cooked meat produce. The tumor initiation activity of PhIP and MeIQx was assayed with Ames method using Salmonella typhimurium TA98 in the presence of S-9 mixtures before and after addition of methanol-solubles of seaweed, such as, Phaeophyta; Undaria pinnatifida, Ecklonia stolonifera Ecklonia cava, Laminaia japonica Sargassum fulvellum, Sargassum hornezi, Sargassum miyabei, Sargassum thunbergii, Agarum cribrosum and Hizikia fusifomis, Rhodophpei Porphyra yezoensis, Grateloupia eiliptiut Lomentraria catenata, Ploemium telfairiae and Glarilaria verrucosa, Chlorophyta; Codium fragile, Enteromorpha compresa and Ulva pertusa. Among seaweed tested, Phaeophyta was shown the higher desmutagenic activity than Rhodophyta and Chlorophyta. E. stolonifera, E. cava and S. miyabei, among Phaeophyta exerted the stronger desmutagenic activity (above $90\%$/2 mg). The ethyl acetate, diethyl ether and chlomform extracts except water extracts from E. stolonifera exhibited a high desmutagenic activity. The ethyl acetate extract of E. stolonifera which showed highest activity was fractionated with Sephadex LH20 column chromatography to give active fraction A-7, which showed desmutagenicity of $90\%$/mg against PhIP and $80\%$/mg against MeIQx. The active fraction had the absorbance at 207.7 and 232nm.

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