• Title/Summary/Keyword: 시료 전처리

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Comparative Studies on the Enzymatic Properties of Trypsins from Cat-shark and Mackerel -1. Purifications and Reaction Conditions of the Trypsins- (복상어와 고등어의 Trypsin에 관한 비교 효소학적 연구 -1. Trypsin의 정제와 반응조건-)

  • PYEUN Jae-Hyeung;CHO Deuk-Moon;HEU Min-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.24 no.5
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    • pp.273-288
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    • 1991
  • To elucidate the physiological and biochemical differences between chondrichthyes and osteichthyes, the properties of the specific digestive enzymes in cat-shark, Cephaloscyllium umbratile, and mackerel, Scomber japonicus, were studied. Homogenous trypsin proved through the disc-electrophoresis, SDS-PAG electrophoresis and gel filtration was obtained from the pancreas of cat-shark by $50-70\%$ saturated ammonium sulphate fractionation, DEAE-Sephadex A-50 column chromatography, benzamidine-Sepharose 6B affinity chromatography and Sephadex G-75-120 gel filtration. Two types of trypsins were also obtained from the pyloric caeca of mackerel by $30-70\%$ saturated ammonium sulphate fractionation and the slightly modified procedure from the method adopted in the purification of cat-shark trypsin. The two trypsins, designated trypsin A and B, were proved their homogeneity by disc- and SDS-PAG electrophoresis and gel filtration. The molecular weights of the trypsins were estimated to be 31,700 for cat-shark trypsin, 30,000 for mackerel trypsin A and 29,000 for mackerel trypsin B by SDS-PAG electrophoresis, but those were estimated to be 21,500 for cat-shark trypsin, 23,700 for mackerel trypsin A and 21,500 for mackerel trypsin B by gel filtration. The trypsins exhibited their optimum conditions at pH 9.0 and on temperature ranged from $45^{\circ}C\;to\;50^{\circ}C$ for cat-shark, and at pH 8.0 and a temperature of $50^{\circ}C$ for mackerel trypsin A and B, respectively. The cat-shark trypsin was stable at pH 10.0 and the temperature below $10^{\circ}C$, whereas the mackerel trypsin A and B, were stable in the range over pH 7.0 to pH 9.0 below $10^{\circ}C$ and at pH 8.0 below $35^{\circ}C$, respectively. The mackerel trypsins were severely inhibited by some heavy metal ions such as $Ag^{2+},\;Cu^{2+}\;and\;Hg^{2+}$ compared to cat-shark trypsin. All of the enzymes were also inhibited by antipain, leupeptin, TLCK(tosyllysine chloromethyl ketone) and SBTI(soybean trypsin inhibitor) remarkably. The inhibitory effects of PMSF(phenylmethane sulphonylfluoride), DFP(diisopropyl fluorophosphate) and benzamidine were indicated that these enzymes belong to serine-proteases.

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Quality characteristics of fresh-cut Dioscorea bulbifera treated under various blanching conditions prior to vacuum-packaging during storage (블랜칭 처리에 의한 신선편이 둥근마의 저장 중 품질 특성)

  • Seong, Bong Jae;Kim, Sun Ick;Jee, Moo Geun;Kim, Soo Dong;Kwon, A Reum;Kim, Hyun Ho;Lee, Ka Soon
    • Food Science and Preservation
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    • v.24 no.5
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    • pp.565-575
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    • 2017
  • This study was carried out to investigate shelf-life and quality of fresh-cut Dungkunma (Dioscorea bulbifera) in order to elevate utilization of Dungkunma a fresh food. Before vacuum-packaging (in polyethylene/polypropylene film ($100{\mu}m$, $15{\times}20cm$, $75{\pm}2cmHg$) and storaging at $2^{\circ}C$, Dungkunma was peeled out and cut to dice type ($2.0{\pm}0.5cm^3$), and then washed and blanched using hot water (at $90{\pm}2^{\circ}C$ with 2% NaCl solution for 30 sec). Blanched Dungkunma was pre-dried at room temperature, $40^{\circ}C$ and $50^{\circ}C$ for removing surface water. Each peeled dice Dungkunma was packed 50 g in polyethylene/polypropylene film ($100{\mu}m$, $15{\times}20cm$) with vacuum treatment ($75{\pm}2cmHg$) and stored at $2^{\circ}C$ for 90 days. Hardness and adhesiveness of Dungkunma blanched by 2% NaCl and pre-dried at $50^{\circ}C$ (SB50) were the highest, but changes were the least during storage. Lightness and yellowness of stored Dungkunma in all treatments decreased slightly while redness increased during storage. Changes of color of SB50 was the least. Total concentration of aerobic bacteria in SB50 was $1.88{\pm}0.18log\;CFU/g$ during 90 days and E. coli was detected in all treatments during whole storage periods. Dioscin and allantoin contents of SB50 were virtually unchanged during the storage. Consequently, the results of this study suggest that vacuum packaged Dungkunma after blanching using 2% NaCl solution could be effective to prolong the quality of fresh-cut Dungkunma.

Compound K (CK) Rich Fractions from Korean Red Ginseng Inhibit Toll-like Receptor (TLR) 4- or TLR9-mediated Mitogen-activated Protein Kinases Activation and Pro-inflammatory Responses in Murine Macrophages (고려홍삼으로부터 분리한 compound K 함유분획에 의한 대식세포의 toll-like receptor-의존성 신호전달로 활성조절 분석)

  • Yang, Chul-Su;Ko, Sung-Ryong;Cho, Byung-Goo;Lee, Ji-Yeon;Kim, Ki-Hye;Shin, Dong-Min;Yuk, Jae-Min;Sohn, Hyun-Joo;Kim, Young-Sook;Wee, Jae-Joon;Do, Jae-Ho;Jo, Eun-Kyeong
    • Journal of Ginseng Research
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    • v.31 no.4
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    • pp.181-190
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    • 2007
  • Compound K (CK), a protopanaxadiol ginsenoside metabolite, was previously shown to have immunomodulatory effects. In this study, we isolated the CK rich fractions (CKRF) from Korean Red Ginseng and investigated the regulation of CKRF-mediated inflammatory signaling during Toll-like receptor (TLR)-mediated cellular activation. Among various TLR ligands, CKRF considerably abrogated TLR4- or TLR9-induced inflammatory signaling. Both LPS and CpG-containing oligodeoxynucleotides (CpG-ODN) stimulation rapidly activates mitogen-activated protein kinases [MAPKs; extracellular signal-regulated kinases 1/2 and p38], NF-${\kappa}B$, and expression of pro-inflammatory cytokines tumor necrosis factor-${\alpha}$, and interleukin-6 in murine bone marrow-derived macrophages (BMDMs) in a time- and dose-dependent manner. Of interest, pre-treatment of CKRF in either LPS/TLR4- or CpG-ODN/TLR9-stimulated macrophages substantially attenuated the LPS-induced inflammatory cytokine production and mRNA expressions, as well as MAPK and NF-${\kappa}B$ activation. To our knowledge, this is the first description of the inhibitory roles for CKRF in TLR4- or TLR9-associated signaling in BMDMs. Collectively, these results demonstrate that CKRF specifically modulates distinct TLR4 and TLR9-mediated inflammatory responses, and further studies are urgently needed for their in vivo roles for potential therapeutic uses, such as in systemic inflammatory syndromes.

Cesium Radioisotope Measurement Method for Environmental Soil by Ammonium Molybdophosphate (환경토양에서 몰리브도인산 암모늄을 이용한 세슘 동위원소 평가방법)

  • Choe, Yeong-hun;Seo, Yang Gon
    • Clean Technology
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    • v.22 no.2
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    • pp.122-131
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    • 2016
  • Caesium radioisotopes, 134Cs and 137Cs which come from the atmospheric nuclear tests and discharges from nuclear power plants, are very important to study artificial radioactivity. In this work, in order to lower the minimum detection activity (MDA) we investigated environmental radioactivity according to the Environment Measurement Laboratory procedure by 137Cs and 134Cs which is similar to chemical and environmental behaviors of 137Cs. The environmental soils in high mountain areas near nuclear power plant were collected, and an Ammonium Molybdophosphate (AMP) precipitation method, which showed high selectivity toward Cs+ ions, was applied to chemically extract and concentrate Caesium radioisotopes. Radioactivity was estimated by a gamma-ray spectrometry. In gamma energy spectrum, with an increasing of 40K radioactivity, it increased the MDA of 134Cs and 137Cs. Therefore, if the natural radionuclides were removed from the soil samples, the MDA of Caesium may be reduced, and the contents of 137Cs of in the environmental soils can effectively be estimated. In the standard soil sample of Korea Institute of Nuclear Safety, radioactivity of 40K was removed more than 84% on average, and the MDA of 134Cs was reduced 2 times. The content of 137Cs was recovered over 84%. On the other hand, in environmental soils, AMP precipitation method showed removal ratio of 40K up to 180 times, which reduced the MDA about 5 times smaller than those of Direct method. 137Cs recovery ratio showed from 54.54% to 70.06%. When considering the MDA and recovery ratio, AMP precipitation method is effective for detection of Caesium radioisotopes in low concentration.

Development of Analysis Method of Caffeine and Content Survey in Commercial Foods by HPLC (HPLC를 이용한 카페인의 분석법 개발 및 시판 식품중 함유량 조사)

  • Kim, Hee-Yun;Lee, Young-Ja;Hong, Ki-Hyoung;Lee, Chul-Won;Kim, Kil-Saeng;Ha, Sang-Chul
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1471-1476
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    • 1999
  • A simple and practical method for determination of caffeine in foods was developed. The analysis of caffeine was performed by reverse phase high performance liquid chromatography using a ${\mu}-Bondapak\;C_{18}$ column at isocratic condition with methanol-acetic acid-water(20 : 1 : 79) on UV detector at 280 nm. The clean-up and extraction of caffeine in samples were based on a simple pretreatment using a Sep-Pak $C_{18}$ cartridge. Recovery rates obtained with this method for cider, candy, cookie, milk, ice cream and persimmon leaf tea were 99.23%, 99.50%, 99.17%, 99.37%, 98.93% and 99.10% respectively. And the detection limit of caffeine was $0.1\;{\mu}g/mL$. With this method, the range of caffeine contents extracted from coffee, green tea, black tea, Oolong tea(tea bag), soft drinks, ice cream, milk and commercial confectionery were $3.38{\sim}37.50\;mg/g,\;16.30{\sim}26.10\;mg/g,\;10.80{\sim}16.65\;mg/g,\;11.25\;mg/g,\;0.06{\sim}0.11\;mg/g,\;0.04{\sim}0.44\;mg/g,\;0.04{\sim}0.39\;mg/g\;and\;0.10{\sim}1.80\;mg/g$, respectively. But caffeine was not detected in the other tea such as Acanthopanax sessiliflorum tea, Angelica gigas tea, Angelica tea, Arrow root tea, Duchu'ng tea, Dunggulle tea, Ganoerma lucidum tea, Ginger tea powder, Persimmon leaf tea, Ssanghwa tea and Cocoa mix powder.

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Determination of plasma C16-C24 globotriaosylceramide (Gb3) isoforms by tandem mass spectrometry for diagnosis of Fabry disease (패브리병(Fabry) 진단을 위한 혈장 중 Globotriaosylceramide (Gb3)의 탠덤매스 분석법 개발과 임상 응용)

  • Yoon, Hye-Ran;Cho, Kyung-Hee;Yoo, Han-Wook;Choi, Jin-Ho;Lee, Dong-Hwan;Zhang, Kate;Keutzer, Joan
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.45-52
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    • 2007
  • Purpose : A simple, rapid, and highly sensitive analytical method for Gb3 in plasma was developed without labor-ex tensive pre-treatment by electrospray ionization MS/ MS (ESI-MS/MS). Measurement of globotriaosy lceramide (Gb3, ceramide trihex oside) in plasma has clinical importance for monitoring after enzyme replacement therapy in Fabry disease patients. The disease is an X-linked lipid storage disorder that results from a deficiency of the enzyme ${\alpha}$-galactosidase A (${\alpha}$-Gal A). The lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly Gb3. Methods : Only simple 50-fold dilution of plasma is necessary for the extraction and isolation of Gb3 in plasma. Gb3 in diluted plasma was dissolved in dioxane containing C17:0 Gb3 as an internal standard. After centrifugation it was directly injected and analyzed through guard column by in combination with multiple reaction monitoring mode of ESI-MS/MS. Results : Eight isoforms of Gb3 were completely resolved from plasma matrix. C16:0 Gb3 occupied 50% of total Gb3 as a major component in plasma. Linear relationship for Gb3 isoforms w as found in the range of 0.001-1.0 ${\mu}g$/mL. The limit of detection (S/N=3) was 0.001 ${\mu}g$/mL and limit of quantification was 0.01 ${\mu}g$/mL for C16:0 Gb3 with acceptable precision and accuracy. Correlation coefficient of calibration curves for 8 Gb3 isoforms ranged from 0.9678 to 0.9982. Conclusion : This quantitative method developed could be useful for rapid and sensitive 1st line Fabry disease screening, monitoring and/or diagnostic tool for Fabry disease.

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Analytical Method for Sodium Polyacrylate in Processed Food Products by Using Size-exclusion Chromatography (Size-exclusion Chromatography를 활용한 가공식품 중 폴리아크릴산나트륨 분석법 확립)

  • Jeong, Eun-Jeong;Choi, Yoo-Jeong;Lee, Gunyoung;Yun, Sang Soon;Lim, Ho Soo;Kim, MeeKyung;Kim, Yong-Suk
    • Journal of Food Hygiene and Safety
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    • v.33 no.6
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    • pp.466-473
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    • 2018
  • An analytical method of sodium polyacrylate in processed food products was developed and monitored by using size-exclusion chromatography. GF-7M HQ column and UV/VIS detector were selected based on peak shape and linearity. Flow rate, column oven temperature, and mobile phase were selected as 0.6 mL/min, $45^{\circ}C$, and 50 mM sodium phosphate buffer of pH 9.0, respectively. Samples for analysis of sodium polyacrylate were extracted with 50 mM sodium phosphate buffer of pH 7.0 for 3 hr at $20^{\circ}C$ and 150 rpm. Analytical method validation revealed proper selectivity and calibration curve was selected in the range of 50-500 mg/L, and correlation coefficient of calibration curve was more than 0.9985. Limit of detection of sodium polyacrylate was 10.95 mg/kg and limit of quantification was 33.19 mg/kg. Accuracy and coefficient of variation for sodium polyacrylate analysis was 99.6-127.6%, 3.0-8.3% for intra-day and 94.3-121.9%, 1.3-2.6% for inter-day, respectively. Sodium polyacrylate was detected in 40 samples among monitored 125 processed food products. Detected contents were less than 0.2%, limited by the Food Additives Code. Results suggest the established size-exclusion chromatography method could be used to analyze sodium polyacrylate in processed food products.

Monitoring of Pesticide Residues in Commonly Consumed Medicinal Agricultural Products (다소비 식·약공용농산물의 잔류농약 실태조사)

  • Hee-Jeong Choi;Yun-Sung Kim;Sang-Tae Kim;Nan-Joo Park;Yu-Mi Choi;Na-Young Yoo;Yoo-Li Han;Jeong-Hwa Seo;Jong-Sung Son;Myoung-Ki Park
    • Journal of Food Hygiene and Safety
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    • v.38 no.3
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    • pp.112-122
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    • 2023
  • We assessed the pesticide residues in the medicinal agricultural products distributed in Korea. Pesticide residues in 72 samples were analyzed using the QuECheRS method for 339 pesticides, using GC-MS/MS and LC-MS/MS. The pesticide residues were below the maximum residues limit (MRL) in 42 samples (58.3%); however, they exceeded the MRL in 5 samples (6.9%). These included 43 types of pesticides; 20 fungicides and 23 insecticides were detected 75 times and 58 times, respectively. Tebuconazole and carbendazim were the most detected fungicides, each 11 times. The levels of acetamiprid, cadusafos, chlorpyrifos, flubendiamide, fluopyram, and triazophos exceeded the MRL in Gogi berry, Omija, and Reishi mushroom. All of them were positive list system (PLS) items that lacked pesticide residue standards. Therefore, authorities should monitor the distributed medicinal agricultural products.

A Study on the analysis method and composition characteristics of organic materials in the pottery excavated at the palace site in Yongjangseong Fortress, Jindo (진도 용장성 왕궁지 출토 도기호 내부 유기물의 분석법과 성분 특성 연구)

  • YUN Eunyoung;YU Jia;KIM Kyuho
    • Korean Journal of Heritage: History & Science
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    • v.56 no.3
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    • pp.158-171
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    • 2023
  • Pottery filled with organic materials was excavated from the G-2 building site of Yongjangseong Fortress, Jingo, a relic of the Goryeo Dynasty. In this study, the characteristics of organic material were confirmed by a scientific analysis of organic material in pottery found at the palace in Yongjangseong, Jindo. In addition, it was intended to review the analysis method to identify the natural resin and to secure characteristic components(biomarkers) for each natural resin and use them as basic data in the future. The organic materials in the pottery were analyzed using attenuated total reflectance Fourier-transformed infrared spectroscopy(ATR-FTIR) and gas chromatography mass spectrometry(GC-MS). The infrared spectral characteristics were estimated to be natural resin, and biomarkers of organic materials were identified as sesquiterpene-based compounds(C15H24, MW 204) and derivatives. The lacquer(T.vemicifluum) is composed mainly of alkenes, alkanes, and catechol. Pine resin(P.densiflora), on the other hand, is primarily composed of diterpenoid(abietic acid, pimaric acid) and Whangchil(yellow lacquer) is identified to have sesquiterpenes(such as selinene, muurolene, calamenene) as its main components. So, the organic material in the pottery can be identified as Whangchil by comparing their compounds with modern resin materials from Dendropanax. morbifera that correspond with the results. Whangchil, which is exuded from the Dendropanax. morbifera, has been used as a natural coating materials since ancient times, and it has been confirmed that the characteristic components are well preserved even 700 years later. It can be assumed that the interior Whangchil was stored not for use as a coating, but rather for ritual purposes when the building was constructed, because the pottery was found near the cornerstone. Furthermore, based on simplified sample preparation using pyrolysis-gas chromatography mass spectrometry(Py-GC-MS), the thermal decomposition products were found to be similar to the characteristic components, suggesting that this method can be applied to the identification of natural resins used in historic artifacts.

A Survey on Pesticide Residues and Risk Assessment for Agricultural Products Marketed in the Northern Area of Seoul from 2022 to 2023 (서울 북부지역 유통 농산물의 잔류농약 실태 및 위해성 평가 (2022-2023))

  • Boram Kwak;Sung ae Jo;Kyeong Ah Lee;Sijung Kim;Yunhee Kim;HyeJin Yi;Seoyoung Kim;Ae Kyung Kim;Eun Sun Yun
    • Journal of Food Hygiene and Safety
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    • v.39 no.3
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    • pp.239-249
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    • 2024
  • In this study, we investigated the concentrations of pesticide residues in agricultural products marketed in northern Seoul and assessed their potential health risks. A total of 1,737 samples were collected using the QuEChERS method, followed by LC-MS/MS and GC-MS/MS. Pesticide residues below the MRLs were detected in 560 samples of 72 items (32.2%), but 38 samples of 22 items had pesticide residues above the MRLs. Residual pesticides were detected in 53.8% of fruits, 33.0% of vegetables, 28.6% of herbs, 15.4% of beans, and 10.5% of rice samples. Most of the samples that exceeded the MRLs were vegetables, especially leafy, stalk, stem, and root vegetables; herbs and mushrooms also exceeded the permitted MRLs. Of the 105 pesticides investigated, dinotefuran, fluxametamide, chlorfenapyr, azoxystrobin, and carbendazim were the most frequently detected, whereas 23 pesticide residues, including terbufos, carbendazim, and fluxametamide, were detected above the MRL values. The hazard indices were calculated as 0.00003-1.31406%, which suggests that the investigated pesticide residues in the samples were within safe levels, but continuous monitoring of pesticides in agricultural products is needed to ensure the safety of consumers.