• Title/Summary/Keyword: Liquid Liquid Extraction

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Purification of Two Novel Antimicrobial Peptides from Pyloric Caeca of the Starfish Asterina pectinifera (별불가사리 Asterina pectinifera의 유문맹낭 추출물로부터 새로운 2종류의 항균활성 펩타이드의 정제)

  • Go, Hye-Jin;Bae, Yun Jung;Park, Nam Gyu
    • Journal of Life Science
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    • v.24 no.8
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    • pp.860-864
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    • 2014
  • PAP-1, a novel antimicrobial peptide isolated from pyloric caeca extract of the starfish Asterina pectinifera was purified and characterized. First, the acidified pyloric caeca extract was put through Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient. Among the eluents, RM 60 (retained materials at 60% methanol) showed good antimicrobial activity against Bacillus subtilis and Escherichia coli D31 and was purified in C18 reversed-phase and ion-exchange high-performance liquid chromatography columns. The purification steps yielded two novel peptides showing strong antimicrobial activities. These peptides were named pyloric caeca A. pectinifera peptide 1 and 2 (PAP-1 and PAP-2). For the characterization of the purified peptides, the molecular weights and amino acid sequences were determined by MALDI-TOF MS and Edman degradation. The molecular weights of PAP-1 and PAP-2 were about 2951.54 Da and 2980.15 Da respectively. The amino acid sequences of PAP-1 and PAP-2 were partially determined: AIQNAGES and AIQNAAES, respectively. PAP-2 is an isoform of PAP-1, differing merely by a single residue at position 6 (glycine or alanine). The comparison of the N-terminal amino acid sequences and molecular weights of the peptides with those of other known antimicrobial peptides revealed that PAP-1 and PAP-2 have no homology with any known peptides. These findings suggest that PAP-1 and PAP-2 play a significant role in the innate defense system of starfish pyloric caeca.

Determination of Synthetic Food Colours by HPLC with Photodiode Array Detector (HPLC를 이용한 타르색소의 분리정량)

  • Yang, Ho-Chul;Heo, Nam-Chil
    • Korean Journal of Food Science and Technology
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    • v.31 no.1
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    • pp.30-35
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    • 1999
  • A simple, rapid, efficient method is for extraction of 13 synthetic water-soluble food colours (Tartrazine, Amarnth, Indigo carmine, New coccine, Sunset yellow FCF, Allura red AC, Eosine, Fast Green FCF, Brilliant Blue FCF, Erythrosine, Acid red, phloxine, Rose Bengal) by polyamide resin and for their quantitative by high performance liquid chromatography (HPLC). Colours (coal-tar dyes) were extracted with polyamide resin and then determinated by HPLC. The HPLC conditions using a reverse phase partition type column $(Nova-pak\;C_{18})$, photodiode array (PDA) detector and 1% Ammonium acetate / 60% acetonitrile in water as eluent, were acceptable for various kinds of colorants. By the use of the proposed method, a survey of coal-tar dyes was carried out on 20 samples and that were detected $4.76{\sim}133.47\;ppm$.

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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.

Studies on the Lipid Components of Potato Tubers - I. Lipid Composition in Free and Bound Lipids - (감자의 지방질(脂肪質) 성분(成分)에 관한 연구(硏究) - 제(第) 1보(報) : 유리(遊離) 및 결합(結合) 지질(脂質)중의 지방질(脂肪質) 조성(組成)에 관하여 -)

  • Lee, Sang-Young;Shin, Hyo-Sun
    • Korean Journal of Food Science and Technology
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    • v.11 no.4
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    • pp.291-297
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    • 1979
  • The experimentally cultivated potatoes of 4 varieties, Irish Cobbler, Warba, Shimabara, and Saco were dried in a frozen state, powdered subsequently and subjected to the extraction of free and bound lipids. Constituents of the prepared lipids were fractionated, quantified, and compared by the methods of column and gas-liquid chromatographies. The results were summarized as follows : 1. The total crude lipid content in potato on a dry weight basis was 0.57 % of which 0.2 % was free lipid and 0.37 % was bound lipid. 2. The neutral lipid content in the free lipid was 14.9 %, approximately 3 times as much as the 4.5 % contained in the bound lipid, whereas the glycolipid content in the free lipid was 15.1 %, slightly less than 22.2 % contained in the bound lipid. However, the phospholipid content was 33.9 % in the bound lipid, approximately 4.5 times as much as the 7.2 % contained in the free lipid. This fact revealed that the bound lipid consisted mainly of polar lipid, while the free lipid consisted of neutral lipids, glycolipids and phospholipids in about the same proportion. 3. The main fatty acids constituting more than 90 % in the free and bound lipids were linoleic, palmitic and linolenic acids. The content of the saturated fatty acid was slightly less in the free lipid than in the bound lipid, whereas the unsaturated fatty acids were more abundant in the free lipid.

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Effect of Enzymatic Deacetylation of T-2 Toxin on the Analysis of T-2 and HT-2 Toxins in Corn and Brown Rice (옥수수 및 현미에서 효소적 탈아세틸화가 T-2와 HT-2 독소 분석에 미치는 영향)

  • Lee, Su-Jin;Ha, Sang-Do;Chun, Hyang-Sook
    • Korean Journal of Food Science and Technology
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    • v.44 no.4
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    • pp.460-466
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    • 2012
  • Through an analysis of T-2 and HT-2 toxins in corn and brown rice, the effect of enzymatic deacetylation of T-2 toxin on HT-2 toxin was investigated. Gas chromatography (GC) with electron capture detection and high-performance liquid chromatography (HPLC) with fluorescence detection were used for quantitative determination. T-2 toxin was converted into HT-2 (84-86%) within 15 min in the presence of crude protein extracts from corn and brown rice. The absence of T-2 conversion was observed for autoclaved samples, in which the enzymes were inactivated. When phosphate buffered saline, followed by methanol, was used as the extraction solvent, recoveries of T-2 toxin spiked at 50 and 200 ${\mu}g/kg$ were from 60 to 87%, whereas those of HT-2 in the autoclaved samples were 0%. In non-autoclaved samples, recoveries of HT-2 were 37-66%, whereas those of T-2 were negligible. However, the conversion of T-2 into HT-2 was not observed when samples were extracted by methanol/water.

Purification and Biological Activities of Bombesin Like Immunoreactivity from Skin of the Frog, Bombina orientalis in Korea (한국산 무당개구리 피부에 존재하는 Bombesin 유사면역 반응물질의 순수정제 및 생물학적 활성)

  • Kwon, Hyeok-Yil;Kim, Yil;Park, Hyoung-Jin
    • The Korean Journal of Physiology
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    • v.24 no.2
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    • pp.363-375
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    • 1990
  • The present investigation was performed to purify bombesin-like immunoreactivity (BBS-LI) from the skin of frogs, B. orientalis inhabiting Korea. For extraction of BBS-LI, the fresh skin of 360 g from frogs was immersed in 1,800 ml of 100% methanol and then kept at $4^{\circ}C$ for 5 days. BBS-LI was partially purified by liquid chromatography using an alkaline alumina column followed by a Sephadex G-10 column. BBS-LI was further purified by using sequential HPLC of reversed phase C18 preparation, gel permeation, SP-ion exchange and reversed phase C18 analysis. BBS-LI in fractions of each step was monitored by radioimmunoassay for which bombesin antiserum with a titer of 1 : 188,800 was raised in a guinea pig. Eventually, two different BBS-LI were successfully purified and each BBS-LI showed the following character. 1) BBS-LI was well separated into two peaks in SP-ion exchange HPLC. One (BBS-LI-K1) bound to the column while the other (BBS-LI-K2) did not. 2) BBS-LI-K1, 73.8% of total BBS-LI, was not differentiated from synthetic bombesin in reversed phase C18 analytical and gel permeation HPLC. 3) BBS-LI-K2, 26.2% of total BBS-LI, eluted later than synthetic bombesin in reversed phase C18 analytical HPLC, but it eluted with a retention time identical to that of synthetic bombesin in gel permeation HPLC. 4) The two forms of BBS-LI and synthetic bombesin identically stimulated gastrin release and pancreatic exocrine secretion including volume, protein output and amylase output in anesthetized rats. It is concluded from the above results that the skin of B. orientalis contains two different forms of BBS-LI which are very identical to bombesin immunologically and biologically. In comparison with synthetic bombesin containing 14 amino acid residues, the major form shows quite similar pattern in all HPLC used in the present study, but the minor form exhibits quite different pattern in SP-ion exchange and reversed phase C18 analytical HPLG.

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Development and validation of analytical methods for pyrifluquinazon residues determination on agricultural commodities by HPLC-UVD (HPLC-UVD를 이용한 농산물 중 pyrifluquinazon 잔류시험법 개발 및 검증)

  • Do, Jung-Ah;Kwon, Ji-Eun;Kim, Mi-Ra;Lee, Eun-Mi;Kuk, Ju-Hee;Cho, Yoon-Jae;Chang, Moon-Ik;Kwon, Kisung;Oh, Jae-Ho
    • Analytical Science and Technology
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    • v.26 no.3
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    • pp.174-181
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    • 2013
  • Pyrifluquinazon is classified with a quinazoline insecticide that regulates food intake by controling the feeding behavior acting on the endocrine or nervous system of pests such as aphids and white fly. To keep safety on pyrifluquinazon residues in agricultural commodities a simple, accurate and rapid analytical method was developed and validated using high performance liquid chromatograph (HPLC-UVD). The pyrifluquinazon residues acidified with 1% formic acid in samples were extracted with acetonitrile and partitioned with hexane subsequently to dichloromethane then purified with silica solid phase extraction (SPE) cartridge. The purified samples were detected using HPLC-UVD. The method was validated using apple and pear spiked with pyrifluquinazon at 0.02, 0.05 and 0.1 mg/kg and hulled rice, pepper, soybean at 0.05 and 0.1 mg/kg. Average recoveries were 70.5~107.9% with relative standard deviation less than 10%. The result of recoveries and overall coefficient of variation of a laboratory results in Gwangju regional FDA and Daejeon regional FDA was followed with Codex guideline (CODEX CAC/GL 40). This method is appropriated at pyrifluquinazon residues determination and will be used as official method of analysis.

Separation of 2,6-dimethylnaphthalene in Dimethylnaphthalene Isomers Mixture by Crystallization Operation (결정화 조작에 의한 Dimethylnaphthalene 이성체 혼합물 중의 2,6-dimethylnaphthalene의 분리)

  • Kang, Ho-Cheol;Kim, Su Jin
    • Applied Chemistry for Engineering
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    • v.25 no.1
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    • pp.116-120
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    • 2014
  • Light cycle oil (LCO), one of the by-products of the catalytic cracking gasoline manufacturing process, contains a lot of valuable aromatics. In particular, 2,6-dimethylnaphthalene (2,6-DMN) contained in LCO has been becoming important as the basic material of polyethylene naphthalate plastic and liquid crystal polymer, etc. If it were possible to separate and purify the valuable aromatic hydrocarbons (such as 2,6-DMN) from LCO, which have only been used as fuel mixed with heavy oil, it would be very meaningful in terms of the efficient use of resources. We investigated the high-purity purification of 2,6-DMN by the combined method of melt crystallization (MC) and solute crystallization (SC). The enriched DMN isomer mixtures (concentration of 2,6-DMN : 10.43%) recovered from LCO by distillation-extraction combination and the crystal recovered by MC used as raw materials of MC and SC, respectively. The solvent of SC used was a mixture of methanol and acetone (60 : 40 wt%). The crystal of 2,6-DMN with a high-purity of 99.5% was recovered by MC-SC combination. We confirmed that the MC-SC combination was one of the very useful combinations for the high-purity purification of 2,6-DMN contained in the enriched DMN isomer mixtures.

Isolation and Characterization of Helicobacter pylori Urease Inhibitor from Rubus coreanus Miquel (복분자(Rubus coreanus Miquel)로부터 Helicobacter pylori Urease Inhibitor의 분리 및 특성)

  • 양성우;호진녕;이유현;신동훈;홍범식;조홍연
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.5
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    • pp.769-777
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    • 2004
  • A Helicobacter pylori urease inhibitor from Rubus coreanus Miquel has been isolated and partially characterized for aiming to Prevent H. pylori growth and decrease harmful accumulation of ammonia in human gastric mucosa. We screened urease inhibitory activities in 519 extracts library prepared by solvent extraction from 173 kinds of edible plants, medicinal herbs, herbs and seaweeds using a colorimetric urease assay system. As results of primary and secondary screening, 70% acetone extract of Rubus coreanus Miquel was selected as potent candidate, showing about 24% inhibitory activity. The acetone extract was sequentially partitioned into RCE/RCWI and RCB/RCW2 layers with ethyl acetate and butanol. The major active component in RCW2, water layer from butanol fractionation was revealed to be peptidic or proteinous substance by inhibitory activity determination after pronase digestion and periodate oxidation. RCW2-IIIc a was isolated by sequential column chromatography on DEAE-Toyopearl 650C, Butrl-Toyopearl 650M and Sephadex LH-20. The isolated urease inhibitor RCW2-IIIc $\alpha$, was highly pure proteinous substance with molecular weight of 13kDa by high-performance gel permeation liquid chromatography. RCW2-IIIc$\alpha$ has about 5 times higher inhibitory activity than 70% acetone extract, showing high stability against heat treatment and peptic digestion.

Antioxidant activities of ethanol extracts from barley sprouts (새싹보리 에탄올 농도별 추출물의 산화방지 활성)

  • Chae, Kyu Seo;Ryu, Eun Hye;Kim, Ki Deok;Kim, Yong-Suk;Kwon, Ji Wung
    • Korean Journal of Food Science and Technology
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    • v.51 no.5
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    • pp.486-491
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    • 2019
  • Phenolic compounds and antioxidant activities of ethanol extracts from barley sprouts were evaluated in this study. Barley sprouts were extracted using water and ethanol in various concentration (25, 50, and 75%) using reflux extraction methods. Ultra performance liquid chromatography (UPLC) analysis showed that barley sprouts are mainly composed of rutin, gallic acid, ferulic acid, and ${\rho}$-coumaric acid. The 75% ethanol extracts had higher total polyphenol contents ($44.01{\pm}1.32mg/g$) and total flavonoid contents ($102.96{\pm}2.49mg/g$). 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity ($EC_{50}$ value: $1.65{\pm}0.02mg/mL$) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical scavenging activity ($EC_{50}$ value: $1.67{\pm}0.02mg/mL$) of the 75% ethanol extracts of barley sprouts were found to be the most effective. The 75% ethanol extracts of barley sprouts exhibited a strong reducing activity and ferric reducing antioxidant activity. As a result, the 75% ethanol extracts of barley sprouts showed stronger antioxidant activity than other extracts.