• Title/Summary/Keyword: Liquid Liquid Extraction

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Purification of TGF-$\beta$ 1 from Human Platelets by an Improved Method (개량된 방법에 의한 사람혈소판으로부터 TGF-$\beta$ 1의 분리)

  • 신충건;김상국;문병조;김평현;전계택;남상욱;김장환;이종원
    • KSBB Journal
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    • v.14 no.1
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    • pp.9-16
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    • 1999
  • Transforming growth factor $\beta$1(TGF-$\beta$1) has potentials to be used as a new therapeutic agent. However, studies with TGF-$\beta$ were hindered by its high cost. In this study, we developed an improved method to purify TGF-$\beta$1 from human platelets, for which four purification steps were used: platelet extraction, gel filtration, cation exchange chromatography, and reverse phase high performance liquid chromatography. After a final step of purification, a pure protein with a molecular weight of 25,000 corresponding to the commercially available TGF-$\beta$1 was obtained, which were confirmed by silver staining and Western blotting after SDS-polyacrylamide gel electrophoresis (SDS-PAGE). It was confirmed by the inhibitory effects of TGF-$\beta$1 on a mink lung epithelial cell line that the purified TGF-$\beta$1 had its biological activity, whose activity is slightly higher than that of the commercially available TGF-$\beta$1. About 3.7$\mug of the purified TGF-$\beta$1 was obtained from 10 units of concentrated human platelets, the final yield of which is about 21%.

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Analysis of Vitamin E in Agricultural Processed Foods in Korea (국내 농산가공식품의 비타민 E 함량 분석)

  • Park, Yeaji;Sung, Jeehye;Choi, Youngmin;Kim, Youngwha;Kim, Myunghee;Jeong, Heon Sang;Lee, Junsoo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.5
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    • pp.771-777
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    • 2016
  • Accurate food composition data are essential for calculation of nutrient intake of a population based on its consumption statistics. In the Korean food composition database, there is a lack of reliable analytical data for tocopherols and tocotrienols. Therefore, this study was conducted to provide information on contents on vitamin E in agricultural processed foods in Korea. Tocopherols and tocotrienols were determined by the saponification extraction method followed by high performance liquid chromatography. Analytical method validation parameters were calculated to ensure the method's validity. Samples were obtained in the years of 2013 and 2014 from the Rural Development Administration. The samples included 34 grains and grain products, 14 snacks, 25 fruits, 5 oils, and 11 sources and spices. All vitamin E isomers were quantitated, and the results were expressed as ${\alpha}$-tocopherol equivalent (${\alpha}-TE$). ${\alpha}-TE$ values of cereal and cereal products, snacks, fruits, oils and sauces and spices ranged from 0.03 to 17.53, 1.01 to 12.84, 0.01 to 1.52, 1.09 to 8.15, and 0.01 to $27.53{\alpha}-TE/100g$, respectively. Accuracy was close to 100% (n=3). Repeatability and reproducibility were 2.04% and 4.69%, respectively. Our study provides reliable data on the tocopherol and tocotrienol contents of agricultural and processed foods in Korea.

Isolation and Structural Analysis of Acetyl Soyasaponin $A_1$ from Hypocotyl of Soybean (콩 Hypocotyl에서 Acetyl Soyasaponin $A_1$의 분리 및 구조 분석)

  • Kim, Sun-Lim;Bang, Myun-Ho;Kim, Jung-Tae;Chi, Hee-Youn;Chung Ill-Min;Kim, Hyun-Bok;Berhow Mark A.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.166-173
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    • 2006
  • Soyasaponins are phytochemicals of major interest fur their health benefits. Chemical investigation of a soybean phytochemical concentrate resulted in the isolation and identification of triterpenoid saponins. The MeOH extraction of defatted hypocotyl separated from soybeans was peformed by the automated solvent extractor (ASE). Fractionation was performed on a flash column ($150mm{\times}40mm$ i.d.) packed with a preparative $C_{18}$ reverse phase bulk packing material $(125\AA,\;55-105{\mu}m)$ and monitored at 210 nm, and collected 14 fractions. Consequent Fsat preparative column liquid chromatography (Fast PCLC) was performed for the purification of Fraction-I (Fr-I) collected from the fraction 8 and 9 of flash chromatography. Fsat PCLC was performed on a Luna $C_{18}\;10{\mu}m,\;100{\AA}$, semipreparative reverse phase column ($250cm{\times}50mm$ i.d.) for the purification of isolated unknown compound (Fr-I-2). Chemical structure of acetyl soyasaponin $A_1\;(MW:1436.6,\;C_{67}H_{104}O_{33})$ was identified and determined by a combination of extensive NMR ($^1H-NMR$, 400 MHz; $^{13}C-NMR$, 100 MHz; DEPT), IR, UV, and ESI-MS analysis.

Preparation and Characterization of Phytochemical-Rich Extract from Sasa quelpaertensis Leaf (식물화합물 다량 함유 제주조릿대 잎 추출물의 제조와 특성)

  • Lee, Ju Yeop;Ko, Hee Chul;Jang, Mi Gyeong;Kim, Se Jae
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1330-1335
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    • 2016
  • Sasa species leaves have been used in traditional medicine for their anti-inflammatory, antipyretic, and diuretic properties. Sasa quelpaertensis Nakai is a small bamboo grass that grows only on Mt. Halla on Jeju Island, Republic of Korea. This small bamboo grass has recently been the focus of much attention due to its potential biomass as well as its beneficial health effects. In this study, to promote the efficient utilization of the S. quelpaertensis leaf, we established a simple preparation method for phytochemical-rich extract (PRE) by comparing phytochemical contents and biological activities according to extraction methods. high performance liquid chromatography (HPLC) analysis revealed that the contents of two major phytochemicals such as, tricin (5.35 mg/g) and p-coumaric acid (44.10 mg/g) contained in PRE were higher than those in fresh hot water extract (SQH, p-coumaric 23.39 mg/g, tricin 0.18 mg/g) and ethanol extract (SQE, p-coumaric 10.8 mg/g, tricin 0.38 mg/g). The antioxidant activities [1,1-diphenyl-2-picrylhydrazy (DPPH) radical scavenging activity, 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity, nitric oxide (NO) scavenging activity, and xanthine oxidase inhibitory activity] of PRE were higher than those of SQH and SQE. PRE effectively inhibited NO production in LSP- stimulation RAW 264.7 cells, and the growth of human promyelocytic leukemia (HL-60) cells. These results suggest that PRE has a potential as a promising antioxidant and anti-inflammatory agent.

Antioxidant Activity of Ripe Fruit Components of Rubus coreanus: Extraction Using Porous Polymer Resins (다공성 합성흡착제를 이용한 복분자 딸기(Rubus coreanus) 농축액의 항산화 효과)

  • Choi, Se-Jin;Lee, Yeon-Sil;Kim, Jin-Kyu;Chung, Cha-Kwon;Kang, Il-Jun;Lim, Soon-Sung
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.149-155
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    • 2011
  • This study was carried out to develop a method of removing unnecessary sugars and high-molecular weight soluble components from water extract by using various highly porous polymer resins: these resins are widely used as adsorbents for polyphenolic compounds. Each anthocyanin-rich fraction (ARF) that was eluted from column packed with 4 different resins (SEPABEADS SP207, 700, 850 and Diaion HP 20) was obtained from an aqueous extract of the ripe fruits of Rubus coreanus, which is well-known in Korea as "Bok-bun-ja". Among the above mentioned resins, Diaion HP20 had the highest efficacy and provided maximum yield of the ARF. The ARF eluted from the column packed with HP 20 exhibited the strongest antioxidant activity in vitro. Bioactive phytochemicals extracted from Rubus coreanus were investigated using the on-line HPLC-$ABTS^{{\cdot}+}$ system, and the resulting 6 peaks were identified as radical-scavenging components. By using liquid chromatography-mass spectrometry, 5 peaks were obtained, and these were identified as cyanidin-3-sambubioside, cyanidin-3-glucoside, cyanidin-3-xylosylrutinoside, cyanidin-3-rutinoside and pelargonidin-3-rutinoside. These results indicate that the use of appropriate porous resin (Diaion HP 20) leads to an increase in the yield of bioactive components and enhancement of their biological properties.

The Anti-bacterial Activity of Eco-friendly Farming Material based on Chinese Nut-gall Extraction on Acidovorax citrulli (오배자 추출물 유래 친환경제제의 세균성과실썩음병균에 대한 항세균활성)

  • Seo, Tae-Jin;Yang, Soo-Jeong;Lee, Bong-Choon;Kim, Kang-Min;Lee, Kui-Jae;Ju, Ho-Jong
    • Korean Journal of Organic Agriculture
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    • v.24 no.3
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    • pp.571-582
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    • 2016
  • Bacterial fruit blotch (BFB) is one of most important diseases in Cucurbitaceae due to infection of Acidovorax citrulli, causing huge economic losses damage worldwide. This seedborn disease spread rapidly at period of high temperature and humidity. The eco-friendly farming is getting popular. So far there was no effective agent to control BFB in eco-friendly farming. Therefore, effect of the material based on chinese nut-gall extract with antibacterial activity against BFB to was tested against A. citrulli. Different hosts showed various symptoms of BFB. Liquid formulation among exhibited most effective anti-bacterial activity on A. citrulli. Pot experiment in greenhouse showed the potential as an control agent of BFB in cucurbits. The treatment of material based on chinese nut-gall extract showed the positive effect on survival of the watermelon seedling and on the length of the cucumber seedling treated with A. citrulli. We cautiously conclude that the material based on chinese nut-gall extract used in this study may be good agents against major diseases of cucurbits in the future even though it is require to be tested with more study on field test.

Development of LC-MS/MS Quantitation Method for Ethoxyquin in Fishery Products (수산물 중 에톡시퀸의 LC-MS/MS 정량분석법 개발)

  • Shin, Dasom;Chae, Young-Sik;Kang, Hui-Seung;Lee, Soo-Bin;Cho, Yoon-Jae;Cheon, So-Young;Jeong, Jiyoon;Rhee, Gyu-Seek
    • Journal of Food Hygiene and Safety
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    • v.31 no.6
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    • pp.432-438
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    • 2016
  • Ethoxyquin (EQ, 1,2-dihydro-6-ethoxy-2,2,4-trimethyl-quinoline) is quinoline-based antioxidant used in the animal feed and food industry to protect the raw materials and final products against oxidation. In recent years the use of synthetic antioxidants in fishmeal ingredients carry-over to farmed fish fillets has received increasing attention in food safety. This study was conducted to develop an analytical method to determine EQ in aquatic products. The analytes were confirmed and quantified via liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the positive ion mode using multiple reaction monitoring (MRM). The sample was extracted with 1 N HCl (in case of flatfish extracted with 1 N HCl containing 10% acetonitrile). Then, solid phase extraction (SPE) was used for the cleanup. Standard calibration curves presented linearity with the correlation coefficient ($r^2$) > 0.99, analyzed at 0.005-0.2 mg/kg concentration. The developed method was validated according to the Codex Alimentarius Commission (CAC) guideline. The limits of quantitation for EQ were 0.01 mg/kg. Average recoveries ranged from 81.3% to 107%. The repeatability of measurements, expressed as the coefficient of variation (CV, %), was below 10%. The analytical method was characterized with high accuracy and acceptable sensitivity to meet CODEX guideline requirements and would be applicable to analyze the EQ residue in aquatic products.

Bitter Peptides Derived from ${\alpha}_{s1}-and\;{\beta}-Casein$ Digested with Alkaline Protease from Bacillus subtilis (Bacillus subtilis의 염기성 프로테아제로 분해된 ${\alpha}_{s1}$- 및 ${\beta}$-카세인에서 분리된 쓴 맛 펩타이드)

  • Sohn, Kyung-Hyun;Lee, Hyong-Joo
    • Korean Journal of Food Science and Technology
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    • v.20 no.5
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    • pp.659-665
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    • 1988
  • The ${\alpha}_{s1}$-and ${\beta}$-casein were purified by DEAE-cellulose chromatography and digested with alkaline protease from Bacillus subtilis. Bitter fractions from the hydrolyzates were isolated using n-butanol extraction, Sephadex G-25 gel chromatography, and high performance liquid chromatography. Peptide mixtures were separated by reverse-phase octadecyl silica column with linear gradient of 0-80% acetonitrile containing 0.1% trifluoroacetic acid. Major peaks were combined from replicate chromatographies and the bitterness of each peak was evaluated. The bitter-tasting peaks were rechromatograpied until isolated peaks were obtained. Three different bitter peptides(BP-I, BP-II, BP-III) were obtained from the ${\alpha}_{s1}$-casein hydrolyzate. BP-I was eluted at 34% acetonitrile and BP-II, 35%, BP-III, 26%, respectively. Two bitter peptides(BP-IV, BP-V) were isolated from the ${\beta}-casein$ hydrolyzate: BP-IV was eluted at 40% acetonitrile and BP-V, 42%. BP-V was the most hydrophobic peptide in the five bitter peptides. However, BP-I and BP-II tasted more bitter than BP-IV and BP-V.

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Simultaneous Determination of Caffeine, Theobromine and Theophylline in Tea Leaves by HPLC (HPLC를 이용한 다류 중 카페인, 테오브로민과 테오필린 동시분석 및 함량 조사)

  • Yun, Sang-Soon;Kim, Hyun;Jang, Su-Jin;Lim, Ho-Soo;Kim, Sheen Hee;Kim, Meehye
    • Korean Journal of Food Science and Technology
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    • v.47 no.5
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    • pp.556-560
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    • 2015
  • This study was aimed at investigating the natural occurrence of caffeine, theobromine, and theophylline in tea leaves. Sample clean-up was based on a simple solid phase extraction (SPE) treatment, and simultaneous analysis was performed by high performance liquid chromatography (HPLC). This method showed good results in terms of linearity, recovery, precision, limit of detection (LOD), and limit of quantitation (LOQ). The caffeine, theobromine, and theophylline contents of tea leaves (n = 228) ranged from 5.4 to 58.2 mg/g, from not detected to 11.2 mg/g, and from not detected to 0.7 mg/g, respectively. The results obtained in this study can be used as fundamental data of caffeine, theobromine, and theophylline contents in tea leaves.

Biokinetics of Carbohydrate and Lipid Metabolism in Normal Laying Hen -II. Incorporation of Activity in Plasma Lipid After Injection of Glucose-U-$C^{14}$- (정상산란계(正常産卵鷄)에 있어서 탄수화물(炭水化物)과 지질대사(脂質代謝)의 생동역학(生動力學) -제2보(第二報) 방사능(放射能)$(C^{14})$포도당 주사후(注射後) 혈장(血漿) 지질(脂質)로의 방사능(放射能)의 병합(倂合)-)

  • Chiang, Y.H.;Riis, P.M.
    • Applied Biological Chemistry
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    • v.21 no.1
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    • pp.63-67
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    • 1978
  • The incorporation of activity in plasma lipid was determined after injection of $u-C^{14}-glucose$ to normal laying hen. The extraction of total lipid from plasma was carried out by the method of Floch et al. with some modifications. The counting of carbon-14 activity was conducted by a Beckman Liquid Scintillation System, The concentration of plasma lipid was estimated as much as 3,070mg per 100ml of plasma taken at 5 minutes after injection of tracer amount of $u-C^{14}-glucose$. The specific activity per gram of plasma lipid carbon atom per injected dose per kg body weight increased gradually following the time after injection until 120 minutes, whereas the glucose activity decreased logarithmically. The partial incorporation quotient of activity in plasma lipid from that of glucose was 0.73% until 120 minutes after the injection.

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