• Title/Summary/Keyword: Extraction method

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Single Laboratory Validation and Uncertainty Estimation of a HPLC Analysis Method for Deoxynivalenol in Noodles (면류에서 HPLC를 이용한 데옥시니발레놀 분석법의 검증과 불확도 산정)

  • Ee, Ok-Hyun;Chang, Hyun-Joo;Kang, Young-Woon;Kim, Mee-Hye;Chun, Hyang-Sook
    • Journal of Food Hygiene and Safety
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    • v.26 no.2
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    • pp.142-149
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    • 2011
  • An isocratic high performance liquid chromatography (HPLC) method for routine analysis of deoxynivalenol in noodles was validated and estimated the measurement uncertainty. Noodles (dried noodle and ramyeon) were analyzed by HPLC-ultraviolet detection using immunoaffinity column for clean-up. The limits of detection (LOD) and quantification (LOQ) were 7.5 ${\mu}g$/kg and 18.8 ${\mu}g$/kg, respectively. The calibration curve showed a good linearity, with correlation coefficients $r^2$ of 0.9999 in the concentration range from 20 to 500 ${\mu}g$/kg. Recoveries and Repeatabilities expressed as coefficients of variation (CV) spiked with 200 and 500 ${\mu}g$/kg were $82{\pm}2.7%$ and $87{\pm}1.3%$% in dried noodle, and $97{\pm}1.6%$ and $91{\pm}12.0%$ in ramyeon, respectively. The uncertainty sources in measurement process were identified as sample weight, final volume, and sample concentration in extraction volume as well as components such as standard stock solution, working standard solution, 5 standard solutions, calibration curve, matrix, and instrument. Deoxynivalenol concentration and expanded uncertainty in two matrixes spiked with 200 ${\mu}g$/kg and 500 ${\mu}g$/kg were estimated to be $163.8{\pm}52.1$ and $435.2{\pm}91.6\;{\mu}g$/kg for dried noodle, and $194.3{\pm}33.0$ and $453.2{\pm}91.1\;{\mu}g$/kg for ramyeon using a coverage factor of two which gives a level of statistical confidence with approximately 95%. The most influential component among uncertainty sources was the recovery of matrix, followed by calibration curve.

Residue analysis of penicillines in livestock and marine products (국내 유통 축·수산물 중 페니실린계 동물용의약품에 대한 잔류실태조사)

  • Song, Ji-Young;Hu, Soo-Jung;Joo, Hyun-Jin;Kim, Mi-Ok;Hwang, Joung-Boon;Han, Yoon-Jung;Kwon, Yu-Jihn;Kang, Shin-Jung;Cho, Dae-Hyun
    • Analytical Science and Technology
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    • v.25 no.4
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    • pp.257-264
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    • 2012
  • Penicillins belong to the ${\beta}$-lactam class of antibiotics, and are frequently used in human and veterinary medicine. Despite the positive effects of these drugs, improper use of penicillins poses a potential health risk to consumers. This study has been undertaken to determinate multi-residues of penicillins, including amoxicillin, ampicillin, oxacillin, bezylpenicillin, cloxacillin, dicloxacillin, and nafcillin, using liquid chromatographic tandem mass spectrometer (LC-MS/MS). The developed method was validated for specificity, precision, recovery, and linearity in livestock and marine products. The analytes were extracted with 80% acetonitrile and clean-up by a single reversed-phase solid-phase extraction step. Six penicillins presented recoveries higher than 76% with the exception of Amoxicillin. Relative standard deviations (RSDs) were not more than 10%. The method was applied to 225 real samples. Benzylpenicillin was detected in 12 livestock products and 7 marine products. Amoxicillin, ampicillin, cloxacilllin, dicloxacillin, nafcillin and oxacillin were not detected. The detected levels were 0.001~0.009 mg/kg in livestock products excluding eggs and milk. In marine products, the detected levels were under 0.03 mg/kg. They were under the MRL levels. As monitoring results, it is identified to be safe but it is considered that safety management of antibiotics should continue by monitoring.

Etiological Properties and Coat Protein Gen Analysis of Potato Virus Y Occuring in Potatoes of Korea (우리나라 감자에 발생하는 PVY의 병원학적 특성 및 외피단백질 유전자 분석)

  • ;Richard M. Bostock
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1995.06b
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    • pp.77-96
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    • 1995
  • To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

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Characteristics of Pop-rice and Rice Tea Using Black Sticky Rice with Giant Embryo (흑찰거대배아미를 이용한 팝라이스와 흑미차의 품질 특성)

  • Han, Sang-Ik;Seo, Woo Duck;Na, Ji-Eun;Park, Ji-Young;Park, Dong-Soo;Cho, Jun-Hyun;Lee, Jong-Hee;Seo, Kyung-Hye;Sim, Eun-Yeong;Nam, Min-Hee
    • Journal of Life Science
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    • v.25 no.1
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    • pp.68-74
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    • 2015
  • Recently developed black waxy rice with a giant embryo ('Nunkeunheukchal', BGE) was selected and processed to produce high quality nutritional food. BGE contains high levels of several phytochemicals with antioxidant activities, as well as other reported health beneficial properties. In addition, the giant embryo has high protein, lipid, and amino acids contents. Within the free amino acids, ${\gamma}$-aminobutyric acid (GABA), a major inhibitory neurotransmitter, has long been used for treating the aftereffects of brain injuries and stroke. A method for manufacturing pop-rice and black rice tea by popping process in BGE is provided to increase a taste, nutrition and functionality. The produced 'pop-rice' showed increased protein (11.3%) and lipid (3.7%) contents compared with control variety, IB ('Ilmibyeo'). In addition, melanoidin related products, polyphenol and functional amino acid contents were increased by the popping process. Pop-rice tea made of BGE showed the highest extraction of total sugar, glucose, raffinose and sucrose (4 times higher than brown rice) by hot water. Scavenging activity ($SC_{50}$) of processed BGE rice powder showed strong antioxidative activity of 0.24 mg/ml using DPPH and 1.82 mg/ml using ABTs method. Thereafter, these results suggested that the popping processed rice of BGE could be one of the promising materials for healthy food development.

Method Development for the Sample Preparation and Quantitative Analysis of Synthetic Colors in Foods (식품 중 식용타르색소의 시료별 전처리방법 확립 및 함량 분석)

  • Park, Sung-Kwan;Lee, Tal-Su;Park, Seung-Kook
    • Korean Journal of Food Science and Technology
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    • v.36 no.6
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    • pp.893-899
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    • 2004
  • Method for sample preparation and quantitative analysis of 19 permitted and non-permitted synthetic colors in foods was developed based on reversed-phase ion-pairing high performance liquid chromatography. For color extraction of samples, deionized water was added, and pH was appropriately adjusted with 1% ammonia water. Any undissolved matters were extracted with 50% ethanol or 70% methanol. Lipid in snacks was first removed using n-hexane with centrifugation, water was added to extract colors, followed by clean-up and concentration using Sep-Pak $C_{18}$ cartridge. Recovery efficiencies at known concentrations of 19 standard food colors spiked into foods were in 90.3-97.9% range far soft drink, 79.2-101.9% for candy, 84.1-103.4% for jelly, 86.4-100.8% for chewing gum, 83.5-103.4% for ice cream, and 78.5-95.6% for snack.

A Criteria on Nitrate Concentration in Soil Solution and Leaf Petiole Juice for Fertigation of Cucumber (Cucumis sativus L.) under Greenhouse Cultivation (시설 오이의 관비재배를 위한 토양용액과 엽병즙액중 질산태 농도 기준 설정)

  • Lim, Jae-Hyun;Lee, In-Bog;Kim, Hong-Lim
    • Korean Journal of Soil Science and Fertilizer
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    • v.34 no.5
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    • pp.316-325
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    • 2001
  • To develope a technique for efficiently managing fertilizer for cucumber, a quick test method to quantify nitrate content in soil solution and leaf petiole juice using a simple instrument was investigated. Among the nitrate analyzing instruments such as compact ion meter, nitrate ion meter, and test strip with reflectometer, the paper test-strip used in conjunction with a hand-held reflectometer was most closely correlated with ion chromatography method in nitrate content, and then it would be suggested with a tool that a farmer can use rapidly, conveniently and accurately for nitrate analysis in a field. Nitrate content in soil solution collected by porous cup was very variable on the lapsed time after drip irrigation and the sampling positions such as soil depth and the distance from dripper. As a result, a significant correlation between nitrate contents of soil solutions and 2M KCl soil extract was not found. However, nitrate content in soil solution extracted with a volume basis (soil:water=1:2) showed the highly significant correlation with that in 2M KCl extract. Nitrate contents of cucumber leaf petiole juices was greatly different between upper and lower leaves. Eleven to sixteen positioned-leaf would be a proper sampling position to determine nitrate content in leaf petiole for evaluating nutrient state by plant tissue analysis. From the secondary regression equations between nitrate contents of soil and petiole juice and the yield of cucumber, nitrate levels for real time diagnosis were estimated as $400mg\;l^{-1}$ soil solution by porous cup. $300mg\;l^{-1}$ in a soil volume extraction, and $1400mg\;l^{-1}$ in petiole juice from spring to summer season. In addition, the maximum yield of cucumber fruit in pot test was obtained in nitrate $1500mg\;l^{-1}$ level of petiole juice, which was similar to nitrate $1400mg\;l^{-1}$ in greenhouse trial.

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Monitoring of Pesticides in the Yeongsan and Seomjin River Basin (영산강 및 섬진강 수계 중 농약 분포 조사)

  • Lee, Young-Jun;Choi, Jeong-Heui;Kim, Sang Don;Jung, Hee-Jung;Lee, Hyung-Jin;Shim, Jae-Han
    • Korean Journal of Environmental Agriculture
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    • v.34 no.4
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    • pp.274-281
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    • 2015
  • BACKGROUND: A lasting release of low levels of persistence chemicals including pesticides and pharmaceuticals into river has a bad influence on aquatic ecosystems and humans. The present study monitored pesticide residues in the Yeongsan and Seomjin river basins and their tributaries as a fundamental study for water quality standard of pesticides.METHODS AND RESULTS: Nine pesticides(aldicarb, carbaryl, carbofuran, chlorpyrifos, 2,4-D, MCPA, methomyl, metolachlor, and molinate) were determined from water samples using SPE-Oasis HLB(pH 2) and LC/MS/MS. Validation of the method was conducted through matrix-matched internal calibration curve, method detection limit(MDL), limit of quantification(LOQ), accuracy, precision, and recovery. MDLs of all pesticides satisfied the GV/10 values. Linearity(r2) was 0.9965- 0.9999, and a percentage of accuracy, precision, and recovery was 89.4-113.6%, 3.1-14.0%, and 90.8-106.2%, respectively. All pesticides exclusive of aldicarb were determined in the river samples, and there was a connection between the positive monitoring results and agricultural use of the pesticides.CONCLUSION: Monitoring outcomes of the present study implied that pesticides were a possible non-point pollutant source in the Yeongsan and Seomjin river basins and tributaries. Therefore, it is required to produce and accumulate more monitoring results on pesticides in river waters to set water quality standards, finally to preserve aquatic ecosystems.

Combination of Enrichment and PCR in Rapid Semi-Quantification of Bacillus cereus in Fresh-Cut Vegetables

  • Choi, Yukyung;Lee, Sujung;Yoon, Yohan
    • Journal of Food Hygiene and Safety
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    • v.35 no.4
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    • pp.319-325
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    • 2020
  • Widespread consumption of fresh-cut vegetables without cooking results in ingestion of major foodborne pathogens including Bacillus cereus. In this study, we aimed to develop a method to rapidly detect B. cereus in fresh-cut vegetables by combining commercial PCR analysis with enrichment of the pathogenic levels. A mixture of B. cereus strains (KCTC1013, KCTC1014, KCTC1092, KCTC1094, and KCTC3624) was inoculated on the surface of fresh-cut cabbage lettuce (20 g) and baby leafy vegetables (10 g) to concentration 1, 2, 3, 4, and 5 log CFU/g. Eighty milliliters of TSB with 0.15% polymyxin B was used for cabbage lettuce, and 90 mL of medium was used for baby leafy vegetables and incubated at 42℃ for 0, 2, 3, 4, 5, 6, and 7 h. One milliliter of the enriched media was plated on mannitol-egg yolk-polymyxin agar for quantification, and another 1 mL was used for DNA extraction for PCR analysis. Additionally, the minimum number of sub-samples to be tested from a pack of fresh-cut vegetable samples was determined using 5 sub-samples. The results from this study showed that for detecting B. cereus in fresh-cut cabbage lettuce, 3, 4, 5, 6, and 7 h enrichment were required to at least detect 5, 4, 3, 2, and 1 log CFU/g of B. cereus, respectively. B. cereus in fresh-cut baby leafy vegetables could be detected after 2, 3, 4, 5, and 6 h of enrichment at 5, 4, 3, 2, and 1 log CFU/g, respectively, using a combination of enrichment and PCR analysis. To determine if a pack of fresh-cut vegetable is positive, the minimum number of sub-samples should be 3. These results can be used to develop a rapid detection method to semi-quantify B. cereus in fresh-cut vegetable samples combining enrichment and PCR.

High Yield Bacterial Expression and Purification of Active Cytochrome P450 p-coumarate-3-hydroxylase (C3H), the Arabidopsis Membrane Protein (대장균 시스템을 이용한 Arabidopsis 막 단백질 cytochrome P450 p-coumarate-3hydroxylase (C3H) 활성형의 과발현 및 분리정제)

  • Yang, Hee-Jung;Kim, Wan-Yeon;Yun, Young-Ju;Yoon, Ji-Won;Kwon, Tae-Woo;Youn, Hye-Sook;Youn, Bu-Hyun
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1039-1046
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    • 2009
  • The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.

Effects of Zizyphus jujuba var. boeunesis Extracts on the Growth of Intestinal Microflora and Its Antioxidant Activities (대추 추출물이 장내 미생물의 생육에 미치는 영향 및 항산화 활성)

  • Jeong, Hye-Mi;Kim, Yi-Seul;Ahn, Seung-Joon;Auh, Mi-Sun;Ahn, Jun-Bae;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.4
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    • pp.500-508
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    • 2011
  • This study was carried out to investigate the effects of jujube extracts on intestinal microflora, along with their antioxidant activities, according to extraction method. The antimicrobial activities of the extracts were measured using the agar diffusion method with a jujube extract concentration of 50 mg/mL. Neither the first nor second jujube extracts were inhibitory against the tested intestinal bacteria. However, water extracts of jujube significantly enhanced the growth of lactic acid bacteria, especially Bifidobacterium bifidum and Bifidobacterium adolescentis. Total phenol compounds and flavonoid compounds were higher in the 1st than in the 2nd water extracts. The EDA values of both water and ethanol extracts increased in proportion to the extract concentration. The 1st water extract showed the highest value among all the others, which was 85.60% at the concentration of 0.05 mg/mL. Furthermore, the 1st water extract showed stronger antioxidant activity than the other samples with an activity of 679.91 mg AA eq/g. These results support the potential use of jujube water extracts as a functional food component and a valuable resource for the development of nutraceutical foods, to increase the growth of Bifidobacterium spp. in the human intestine.