• Title/Summary/Keyword: methanol extraction

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Multiresidue matrix solid phase dispersion (MSPD) extraction and HPLC determination of tetracyclines in animal muscle tissue (시료고체상분산(matrix solid phase dispersion)전처리법을 이용한 식육중 테트라사이클린계 항생물질 동시정량분석)

  • Kang, Hwan-goo;Son, Seong-wan;Cho, Byung-hoon;Lee, Hye-sook;Park, Shin-ja;Kim, Jae-hak;Cho, Myung-haing
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.541-550
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    • 1996
  • Tetracycline antibiotics have been widely used not only therapeutics but feed additives. There are many methods for the isolation and determination of tetracycline antibiotics in animal muscle tissue. But those methods take much time and labor, so it is difficult to analyse many samples simultaneously. A rapid isolation method and liquid chromatographic determination of tetracycline antibiotics in animal muscle tissue (bovine, porcine, chicken) is presented. Blank control and tetracyclines fortified samples (0.5g) were blended with $C_{18}$ containing 0.05g each of oxalic acid and disodium ethylenediaminetetraacetate. After homogenize, homogenate was transferred to glass column made from 10ml glass syringe and compressed to 4~4.5ml volume. A column made from the $C_{18}$/meat matrix was washed with hexane (8ml) and dichloromethane (8ml, if needed), following which the tetracyclines were eluted,vith methanol or 0.01M methanolic oxalic acid (8ml). The eluates containing tetracyclines analytes were free from interfering compounds when analysed by HPLC with UV detection (photodiode array at 360nm). Standard curve for each tetracycline showed a linear response at the range of $0.05{\sim}1.0{\mu}g/ml$ and tetracycline antibiotics were eluted within 4ml of eluted volume. All tetracycline antibiotics except tetracycline were stable during the concentration process at $40^{\circ}C$ and time required for concentration was 3~4 hours. Fortified samples containing oxalic aicd and EDTA represented more good recoveries than those of not-contained sample. Recoveries were 91.8~110.1% (oxytetracycline; OTC), 57.7~79.5% (tetracycline; TC), 78.1~88.6% (chlortetracyclines; CTC) and 88.4~100.6% (doxycycline; DC) in pork tissue, 101.1~126.8% (OTC), 66.4~75.4% (TC), 79.2~88.1% (CTC) and 69.3~86.7% (DC) in beef tissue, and 90.8~95.6% (OTC), 66.2~84.4% (TC), 75.7~77.2% (CTC) and 55.6~80.7% (DC) in chicken muscle tissue. The detection limits validated in muscle tissue by this method were $0.05{\mu}g/g$ for OTC and TC, and $0.1{\mu}g/g$ for CTC and DC.

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A Simple and Fast Microplate Method for Analysis of Carotenoids Content in Chili Pepper (Capsicum annuum L.) (고추의 카로티노이드 함량 분석을 위한 신속·간편 마이크로플레이트법)

  • Hong, Sun Chul;Han, Jung-Heon;Lee, Jundae;Ahn, Yul Kyun;Yang, Eun-Young;Chae, Soo Young;Kim, Su;Yoon, Jae Bok
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.807-812
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    • 2013
  • In this study, we aimed to develop a simple analysis method for measuring the carotenoids content of pepper powder. A 96-wells polystyrene microplate and an ELISA reader were used for analysis. Although ELISA reader with 450 nm filter was applicable to measure carotenoid contents, the surface of microplates were degenerated by acetone used for carotenoids extraction. However, ten-folded dilute of the color extract with methanol did not affect the surface of polystyrene microplate and components of the color extract could be successfully measured by a ELISA reader, showing a high corelation with ASTA-20.1 method. In addition, this method uses 10 fold less acetone than ASTA-20.1 method resulting less acetone waste. The microplate method using ELISA reader has potential power for analyzing a large number of samples which may be very useful to the practical breeding program for high-colored peppers.

Determination of Six Sweeteners in Children's Favorite Foods by HPLC-MS/MS (HPLC-MS/MS를 이용한 어린이 기호식품중의 감미료 분석)

  • Kim, Il-Young;Du, Ok-Ju;Lee, Sung-Dck;Park, Young-He;Kim, Mi-Sun;Bea, Chung-Ho;Chae, Young-Zoo
    • Journal of Food Hygiene and Safety
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    • v.25 no.2
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    • pp.118-121
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    • 2010
  • A HPLC-MS/MS method was developed for simultaneous determination of six sweeteners (acesulfame-K, cyclamate, saccharin, sucralose, stevioside, aspartame) in children's favorite foods. The procedure involves an extraction of the six sweeteners with 50% methanol solution, sample clean-up using the Carrez clearing reagent and filtering with cartridge filter. The HPLC separation was performed on a Hypersil Gold (150 mm ${\times}$ 2.1 mm 5 um) column using the water/acetonitrile mobile phase (95:5). Mass spectrometric analysis was carried out using the TSQ Quantum Ultra operated in negative and positive ESI/SRM. With this method, good linear relationship, sensitivity and reproducibility were obtained. The spike recoveries of six sweeteners for 2 kinds of foods spiked into 0.4 mg/ kg ranged from 87.4 to 114.7%. The detection limits were above 0.02 mg/kg. The method has been applied to determination of six sweeteners in children's favorite foods.

Determination of Neonicotinoid Pesticides in Commercial Agricultural Products by LC-MS/MS (LC-MS/MS를 이용한 농산물중 Neonicotinoid 계 농약분석)

  • Hwang, Lae-hong;Yang, Hye-ran;Lee, Jae-kyoo;Kim, Chang-kyu;Kim, Min-jung
    • Journal of Food Hygiene and Safety
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    • v.35 no.4
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    • pp.312-318
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    • 2020
  • A method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for neonicotinoid pesticide analysis in agricultural products. Four compounds (imidacloprid, clothianidin, acetamiprid, thiacloprid) were extracted with acetonitrile from agricultural products and cleaned up by NH2 solid-phase extraction procedure, and eluted with 0.1% formic acid in methanol/dichloromethane (5/95, v/v). The limit of detection and quantification were 0.0001-0.0005 mg/kg and 0.001 mg/kg, respectively. The mean recoveries of neonicotinoid pesticide from agricultural products were in the range of 90.7-100.9% and 94.4-99.8%, as spiked at 0.2 mg/kg and 0.02 mg/kg, respectively. This validation satisfied the national criteria for pesticide analytical methods. In summary, The present method is fast, precise and sensitive enough for the Positive List System (PLS), and we conclude that the method is also suitable for neonicotinoid pesticide determination in a wide range of agricultural products.

Antioxidant Activity of Various Fractions Extracted from Mustard Leaf (Brassica juncea) and Their Kimchi (청갓과 청갓김치의 용매별 추출물의 항산화성)

  • Kim, Jae-I;Park, Jae-Sue;Kim, Woo-Seong;Woo, Kang-Lyung;Jeon, Jung-Tae;Min, Byung-Tae;Cheigh, Hong-Sik
    • Journal of Life Science
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    • v.14 no.2
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    • pp.286-290
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    • 2004
  • The antioxidant activities of the various fractions extracted from mustard leaf (Brassica juncea) and mustard leaf kimchi were determined by the radical scavenging effect on 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical. The fractions from dried mustard leaf, and fermented mustard leaf kimchi for 0 day and fermented mustard leaf kimchi for 5 days at 15$^{\circ}C$ were screened for the scavenging effects by using DPPH assay. The fractions prepared by the systematic extraction procedure with the solvents such as hexane, C $H_2$Cl$_2$, EtOAc, BuOH and $H_2O$ were used for the determination of free radical scavenging effects. The antioxidant activity of EtOAc and n-BuOH soluble fraction from mustard leaf and mustard leaf kimchi for 0 day had stronger than the others. During fermentation at 15$^{\circ}C$ for 5 days, the antioxidant activity was changed. C $H_2$Cl$_2$ and EtOAc soluble fraction showed more potent radical scavenger effects than the others. The difference or results were to the various supplements and fermentation process.

Antioxidative Activity and Chemical Characteristics of Leaves, Roots, Stems and Fruits Extracts from Acanthopanax senticosus (가시오가피 잎·뿌리·추출물의 이화학적 특성 및 항산화 효과)

  • Heo, Su-Jin;Ahn, Hee-Young;Kang, Min-Jeong;Lee, Jae-Hong;Cha, Jae-Young;Cho, Young-Su
    • Journal of Life Science
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    • v.21 no.7
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    • pp.1052-1059
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    • 2011
  • The contents of bioactive and antioxidative activities (DPPH (${\alpha},{\alpha}'$-diphenyl-${\beta}$-picrylhydrazyl), free radical scavenging activity, peroxidation of linoleic acid and rat hepatocyte microsome, and Fe/Cu reducing power, tyrosinase inhibition activity) were tested by in vitro experimental models using water, hot water, ethanol and methanol extracts of leaves (ASL), roots (ASR), stems (ASS) and fruits (ASF) from Acanthopanax senticosus. Hot water extract from ASL showed the highest extraction yield (16.04%) as well as highest contents of phenolic compounds (2.67%) and flavonoids (1.43%). Major minerals were K, Ca and Mg. In oxidation in vitro models using DPPH free radical scavenging activity, Fe/Cu reducing power, $Fe^{2+}$/ascorbate-induced linoleic acid peroxidation by ferric thiocyanate and thiobarbituric acid (TBA) methods, tyrosinase inhibition activity and autooxidation of rat hepatic microsomes membrane, and antioxidative activities were strong in Acanthopanax senticosus. From these results, ASL extracts were shown to have the most potent antioxidative properties and contain the highest amounts of antioxidative compounds such as phenolic compounds and flavonoids. These results may provide the basic data to understand the biological activities of bio-active materials derived from leaves of Acanthopanax senticosus.

Chemical compositions and functional characteristics of Korean and imported pomegranate (Punica granatum L.) (국내산과 수입산 석류의 화학적 성분과 기능적 특성)

  • Kim, Mi Sook;Yun, Seol Hee;Na, Hwan Sik;Park, Hark Jae;Choi, Gyeong Cheol;Yang, Soo In;Lee, Ji Heon
    • Food Science and Preservation
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    • v.20 no.3
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    • pp.342-347
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    • 2013
  • In recent years, polyphenol-rich herbs, fruits and processed foods, which are made of plant origin, have attracted much attention due to their potential health benefits. Pomegranate (Punica granatum L.) is an important source of bioactive compounds and has been used to treat diseases because of its medicinal properties. This research was focused on characterizing Korea's national cultivar and a similar product from California, USA. To evaluate their bioactive compounds and pharmacological activities, their anti-oxidation and cancer inhibition properties, as well as their organic acid and free sugar contents, were investigated. The national cultivar had low total sugar and high organic acid contents, contrary to the imported product. The results showed that the peel of national cultivar had high polyphenol and ellagic acid contents compared to imported product. The free radical scavenging capacity was evaluated via 2,2-diphenyl-1-picrylhydrazyl (DPPH) and its positive correlation with the total polyphenol contents was found. The anti-cancer activity of methanol extracts revealed growth inhibition against the prostate cancer cell. These results signify that while pomegranate, national cultivar, is more sour than the imported product, its health benefits could be excellent. Also, the polyphenol compound content of the non-edible part (such as the peel and the seed) was higher than that of the juice. Thus, it is suggested that the byproduct of the juice extraction could be potentially used in other fields such as medicine or dietary agents.

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.

Bioactive Compounds and Antioxidant Activity of Jeju Camellia Mistletoe (Korthalsella japonica Engl.) (제주 동백나무 겨우살이의 용매별 기능성 성분 및 항산화 작용)

  • Kang, Da Hee;Park, Eun Mi;Kim, Ji Hye;Yang, Jung Woo;Kim, Jung Hyun;Kim, Min Young
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1074-1081
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    • 2016
  • Mistletoes are hemi-parasitic plant growing on different host tree and shrubs. They are traditionally used in folkloric medicine for the treatment of diarrhea, cough, diabetes, hypertension, cancer and skin infection. The purpose of this study was to determine the contents of phenolics and antioxidant activity of 70% ethanol, 100% methanol and hot water extracts of Jeju camellia mistletoe (Korthalsella japonica Engl.). Ethanol was most effective in extracting total phenols (7,427 mg gallic acid equivalent (GAE)/100 g) and flavonoid (1,777 mg rutin equivalent (RE)/100 g). The free radical scavenging activity, 1,1-diphenyl-2-picryl hydrazyl (DPPH) (EC50 = 7.8 mg/ml) and hydrogen peroxide (H2O2) (EC50 = 1.4 mg/ml), and the capacity for chelating metal ions (EC50 = 8.0 mg/ml) and reducing power (EC50 = 14.9 mg/ml) of the samples also higher in ethanolic extracts. The strong correlation (r2 = −0.996~−0.881) between antioxidant capacities and the phenolic contents implied that phenolic compounds are a major contributor to the antioxidant activity of the ethanolic extracts of Jeju camellia mistletoe. As conclusions, Jeju camellia mistletoe contains bioactive substances with a potential for reducing the physiological as well as oxidative stress and this could explain the suggested cancer preventive effect of these plants as well as their protective role on other major diseases.

Isolation of Alliin in Garlic and Its Quantitative Determination by High Performance Liquid Chromatography and Studies on the Antimicrobial Efforts of Alliin and Ethanol Extracts from Korean Garlic(Alliium sativum L.) (마늘 중 고속 액체 크로마토그래피에 의한 알린의 분리 및 정량과 Alliin과 에탄올 추출물의 항균효과에 관한 연구)

  • 위성언
    • The Korean Journal of Food And Nutrition
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    • v.16 no.4
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    • pp.296-302
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    • 2003
  • First. the purification and analysis of alliin in garlic from different origins by alliin-HPLC determination method were studied. Allinase in garlic was inactivated by heating in boiling water followed by extraction of alliin in garlic with 80% methanol. To remove free amino acids and alliin homologs in garlic, garlic extract was separated by cation exchange column which was packed with amberlite CG-120 resin using 40L d-water as eluent. Alliin in garlic extract was crystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=70:29:1 and then recrystallized in a mixture of acetone (50$^{\circ}C$):H$_2$O:acetic acid=75:24:1. Obtained alliin was identified by melting point. TLC, microscope observation and mass spectrometry. High performance liquid chromatography (HPLC) following pre-column derivatization of cystein derivatives with o-phthaldialdehyde/2-mercaptoethanol has succeessfully been applied to the analysis of various garlics. Each alliic of standard solution and garlic extract was derivatized to isoindole derivative by o-phthaldialdehyde /2-mercaptoethanol and then analyzed by HPLC. Six point calibration was done by using alliin peak area. Lineality was observed at 0 ∼ 1.0mg/ml of alliin concentration. Weighted regression line function was Y=6254X - 256077. By this function, alliin contents in various garlics were 0.34 ∼ 0.73% fresh weight. Second study was designed to evaluate the effects of garlic extracts of various concentrations on the growth of various pathogenes (Eubacterium limonsum, Bacteroides fragilis, Salmonella typhimurium, Salmonella typhi, Shigella sonnei, Kiebsiella pneumoniae, Enterobacter cloacae, Pserdomonas aeruginosa, Escherichia coli). For antimicrobial effects against microorganism, totally minimal inhibition concentrations (MIC) of alliin were from 5,000 to 20,000ppm. MIC of ethanol extract were 1,250 to 10,000ppm.