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Development of an Official Analytical Method for Determination of Phorate and its Metabolites in Livestock Using LC-MS/MS (LC-MS/MS를 이용한 축산물 중 Phorate 및 대사산물 5종 동시분석법 개발)

  • Ko, Ah-Young;Kim, Heejung;Jang, Jin;Lee, Eun Hyang;Ju, Yunji;Noh, Mijung;Kim, Seongcheol;Park, Sung-Won;Chang, Moon-Ik;Rhee, Gyu-Seek
    • Journal of Food Hygiene and Safety
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    • v.30 no.3
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    • pp.272-280
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    • 2015
  • A simultaneous official method was developed for the determination of phorate and its metabolites (phorate sulfoxide, phorate sulfone, phorate oxon, phorate oxon sulfoxide, phorate oxon sulfone) in livestock samples. The analytes were quantified and confirmed via liquid chromatograph-tandem mass spectrometer (LC-MS/MS) in positive ion mode using multiple reaction monitoring (MRM). Phorate and its metabolites were extracted from beef and milk samples with acidified acetonitrile (containing 1% acetic acid) and partitioned with anhydrous magnesium sulfate. Then, the extract was purified through primary secondary amine (PSA) and C18 dispersive sorbent. Matrix matched calibration curves were linear over the calibration ranges (0.005-0.5 mg/L) for all the analytes into blank extract with $r^2$ > 0.996. For validation purposes, recovery studies were carried out at three different concentration levels (beef 0.004, 0.04 and 0.2 mg/kg; milk 0.008, 0.04 and 0.2 mg/kg, n = 5). The recoveries were within 79.2-113.9% with relative standard deviations (RSDs) less than 19.2% for all analytes. All values were consistent with the criteria ranges requested in the Codex guidelines. The limit of quantification was quite lower than the maximum residue limit (MRL) set by the Ministry of Food and Drug Safety (0.05 mg/kg). The proposed analytical method was accurate, effective and sensitive for phorate and its metabolites determination and it will be used to as an official analytical method in Korea.

Simultaneous Determination of Penicillin Antibiotics in Meat using Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS를 이용한 육류 중 페니실린계 항생제 8종의 동시분석 및 적용성 검증)

  • Kim, Myeong-Ae;Yoon, Su-Jin;Kim, MeeKyung;Cho, Yoon-Jae;Choi, Sun-Ju;Chang, Moon-Ik;Lee, Sang-Mok;Kim, Hee-Jeong;Jeong, Jiyoon;Rhee, Gyu-Seek;Lee, Sang-Jae
    • Journal of Food Hygiene and Safety
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    • v.29 no.2
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    • pp.131-140
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    • 2014
  • The objective of this study was to develop a simultaneous method of 8 penicillin antibiotics including amoxicillin, ampicillin, cloxacillin, dicloxacillin, nafcillin, oxacillin, penicillin G and penicillin V in meat using LC-MS/MS. The procedure involves solid phase extraction with HLB cartridge and subsequent analysis by LC-MS/MS. To optimize MS analytical condition of 8 compounds, each parameter was established by multiple reaction monitoring in positive ion mode. The chromatographic separation was achieved on a $C_{18}$ column with a mobile phase of 0.05% formic acid and 0.05% formic acid in acetonitrile at a flow rate of 0.2 mL/min for 20 min with a gradient elution. The developed method was validated for specificity, linearity, accuracy and precision in beef, pork and chicken. The recoveries were 71.0~106%, and relative standard deviations (RSD) were 4.0~11.2%. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.003~0.008 mg/kg and 0.01~0.03 mg/kg, respectively, that are below maximum residue limit (MRL) of the penicillins. This study also performed survey of residual penicillin antibiotics for 193 samples of beef, pork and chicken collected from 9 cities in Korea. Penicillins were not found in all the samples except a sample of pork which contained cloxacillin (concentration of 0.08 mg/kg) below the MRL (0.3 mg/kg).

Analysis and Risk Assessment of Benzo(a)pyrene in Edible Oils (식용유지류 중 벤조피렌의 함량 분석 및 안전성 평가)

  • Jang, Mi-Ra;Hong, Mi-Sun;Jung, So-Young;Choi, Bu-Chuhl;Lee, Kyeong-Ah;Kum, Jin-Young;Kim, Il-Young;Kim, Jung-Hun;Chae, Young-Zoo
    • Journal of Food Hygiene and Safety
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    • v.29 no.2
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    • pp.141-145
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    • 2014
  • To assess the health risk for benzo(a)pyrene by the intake of edible oils, 288 cases of edible oils collected from food markets were analysed using the high performance liquid chromatography with fluorescence detector. The levels of benzo(a)pyrene were from non-detection to $4.78{\mu}g/kg$, and the average was $0.11{\mu}g/kg$. The chronic daily exposures of benzo(a)pyrene for total population group and consumer-only group were estimated using the food consumption data in the fifth Korea National Health and Nutrition Examination Survey in 2011. The estimated daily intake of benzo(a)pyrene was $4.26{\times}10^{-3}ng/kg$ b.w./day for total population group and $7.64{\times}10^{-3}ng/kg$ b.w./day for consumer-only group. The MOE (margin of exposure) of benzo(a)pyrene for total population group and consumer-only group was $7.28{\times}10^7{\sim}1.74{\times}10^8$ and $3.95{\times}10^7{\sim}9.42{\times}10^7$, respectively. Accordingly, the health risk from benzo(a)pyrene caused by the intake of edible oils was considered as a very low level.

A Study on the Analysis of Five Artificial Sweetners in Beverages by HPLC/MS/MS (HPLC/MS/MS를 이용한 음료류 중 인공감미료 동시분석에 관한 연구)

  • Lee, Seong-Bong;Yong, Kum-Chan;Hwang, Sun-Il;Kim, Young-Su;Jung, You-Jung;Seo, Mi-Young;Lee, Chang-Hee;Sung, Jin-Hee;Yoon, Mi-Hye
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.327-333
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    • 2014
  • A method for analysis of five artificial sweetners (sodium saccharin, aspartame, acesulfame-K, sucralose, cyclamate) in beverage samples was developed using high-performance liquid chromatography/triple quadrupole mass spectrometry (HPLC/MS/MS). The method uses a single-step dilution for sample preperation. Seperation was achieved on a $C_{18}$ column ($2.1{\times}150mm$, $3.5{\mu}m$) with A- 2% methanol (1 mM ammonium acetate), B-95% methanol (1 mM ammonium acetate) as mobile phase with gradient mode. The quantitation of target compounds was performed by external calibration in selected reaction monitorning (SRM) mode. The coefficient of determination of calibration curve for sodium saccharin, aspartame, acesulfame-K, sucralose and cyclamate were 0.9957, 0.9991, 0.9943, 0.9982 and 0.9948, respectively. The limits of detection (LODs) and limits of quantitation (LOQs) were in the range of 0.001~0.022 mg/L and 0.004~0.073 mg/L, repectively. Recoveries for beverage samples were in the range of 92.76~113.50% with RSD < 10.91%. The method has applied to the determination of the five sweetners in 102 beverage samples. Three artificial sweetners-aspartame, acesulfame-K, sucralose were detected from 42 samples. Sodium saccharin and cyclamate were not detected in all samples.

Analytical Method Development for Determination of Silymarin by LC-MS/MS for Related Health Functional Foods (LC-MS/MS를 이용한 건강기능식품 중 실리마린 분석법 연구)

  • Oh, Mihyune;Lee, Jin Hee;Kim, Sang-A;Kim, Meehye
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.124-130
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    • 2018
  • The Ministry of Food and Drug Safety (MFDS) is amending its test methods for the use of health functional foods (dietary food supplement), in order to establish regulatory standards and specifications in Korea. In this regard, we continue to pursue and perform our research on the analytical method development for the items being researched and reviewed. In this study, we have developed a sensitive and selective test method that could simultaneously separate and determinate six major bioactive flavonolignans in silymarin, which are based on the use of a liquid chromatographic-tandem mass spectrometry (LC-MS/MS). The standard calibration curves presented a linearity effect with the correlation coefficient ($r^2$) > 0.999. The limits of detection (LODs) and limits of quantitation (LOQs) were in the range of $0.3{\sim}9.0{\mu}g/L$ and $0.8{\sim}27.3{\mu}g/L$, respectively. The recovery results ranged between 96.2~98.6% at 3 different concentration levels, and its relative standard deviations (RSDs) were less than 5% as noted in this study. The proposed analytical method was characterized with a noted high resolution of the individual silymarin constituents, and the assay was fully validated as well. Our research can provide a significant scientific evidence that can be useful to amend the silymarin test method for the Health Functional Food Code.

Studies on the Determination Method of Monascus Pigments in Foods (식품 중 홍국색소의 분석법에 관한 연구)

  • Lee, Tal-Soo;Lee, Young-Ja;Kwon, Yong-Kwan;Park, Jae-Seok;Ko, Hyun-Sook;Sim, Kyu-Chang;Lee, Ju-Yeun;Shin, Jae-Wook;Song, Jee-Won;Lee, Chul-Won
    • Korean Journal of Food Science and Technology
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    • v.33 no.5
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    • pp.641-644
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    • 2001
  • This study was performed for development of new analytical method of monascus pigments in foods. In this method, analysis of monascus pigment in foods has been carried out by detection of monascin and ankaflavin of the main color component of monascus pigment as indicator compounds. Monascin and ankaflavin were isolated and identified by TLC, HPLC, Prep. HPLC, $^{1}H-NMR$ and Mass spectrophotometer. The analysis of monascin and ankaflavin in foods such as massal, sausage, mixed press ham, mixed fish sausage, semi-dried sausage and syrup was performed by using reverse phase high performance liquid chromatograph with Capcell Pak C18 column at wave length 390 nm. The quantitative results of monascin were as follows : $0.01{\sim}3.31\;{\mu}g/g$ item in massal, $0.05{\sim}0.10\;{\mu}g/g$ in mixed fish sausage, and $0.34{\sim}0.35\;{\mu}g/g$ in semi-dried sausage. But the quantitative results of ankaflavin were as follows: $0.02{\sim}0.89\;{\mu}g/g$ in massal, ankaflavin were not founded in other samples.

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Contents and Estimated Intakes of Trans Fatty Acids in Korean Diet (한국인의 식품 중 트란스 지방산의 함량과 섭취량 추정)

  • Kim, Jong-Hee;Jang, Kyung-Won;Shin, Hyo-Sun
    • Korean Journal of Food Science and Technology
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    • v.32 no.5
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    • pp.1002-1008
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    • 2000
  • The purpose of this study was to provide the fundamental information for establishing the database needed to estimate total intakes of trans fatty acids in Korea. The amounts of trans fatty acids contained in 164 samples including 25 samples of margarines, 21 samples of shortenings, 19 samples of vegetable salad and cooking oils, 53 samples of confectionery products, 18 samples of bakery products, 19 samples of dairy products, and 9 samples of animal fats and meats were analyzed by capillary gas liquid chromatography. The average amounts of trans fatty acids in those foods were calculated and expressed as gram per one serving. Then, the average daily intakes of trans fatty acids per capita were estimated using the analyzed amounts of trans fatty acids and the amount of yearly production for those foods. The amounts of trans fatty acids per 100 g of lipids were $2.11{\sim}33.83%$ (14.66% on average) in margarines, $1.47{\sim}44.48%$ (14.21% on average) in shortenings, $0.18{\sim}3.82$ (1.54% on average) in vegetable salad and cooking oils, $0{\sim}45.81%$ (10.92% on average) in confectionery products, $0{\sim}18.32%$ (7.87% on average) in bakery products, $0.90{\sim}4.54%$ (2.27% on average) in dairy products, and $0.61{\sim}6.07%$ (2.24% on average) in animal fats and meats. Major isomers of trans fatty acid in the sample foods were $C_{18:1}$ and $C_{18:2}$. As a result, the korean average daily intake of trans fatty acids in korea was estimated to be 2.3 g per capita. The amounts of trans fatty acids consumed from each selected food were as follows: 0.35 g from margarines, 0.57 g from shortenings, 0.11 g from vegetable salad and cooking oils, 0.65 g from confectionery products, 0.07 g from bakery products, 0.14 g from dairy products and 0.21 g from animal fats and meats.

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Laboratory Tests for Trichloroethylene (TCE) and Toluene Remediation in Soil Using Soil Vapor Extraction (토양증기추출(Soil Vapor Extraction)을 이용한 토양 내 Trichloroethylene (TCE)과 Toluene정화 실험)

  • 이민희;강현민
    • Economic and Environmental Geology
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    • v.35 no.3
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    • pp.221-227
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    • 2002
  • Column experiments were performed to evaluate the removal efficiency of soil vapor extraction (SVE) iota TCE (trichloroethylene) and toluene in soil. Homogeneous Ottawa sands and real soils collected from contaminated area were used to investigate the effect of soil properties and SVE operation conditions on the removal efficiency. In column teats with two different sizes of Ottawa sand, the maximum effluent TCE concentration in a coarse sand column was 442 mg/L and 337 mg/L in a fine sand column. However, after 20 liter gas flushing, the effluent concentrations were very similar and more than 90% of initial TCE mass were removed from the column. For two real contaminated soil columns, the maximum effluent concentration decreased 50% compared with that in the homogeneous Ottawa coarse sand column, but 99% of initial TCE mass were extracted from the column within 40 liter air flushing, suggesting that SVE is very available to remove volatile NAPLs in the contaminated soil. To investigate the effect of contaminant existing time on the removal efficiency, an Ottawa sand column was left stable for one week after TCE was injected and the gas extraction was applied into the column. Its effluent concentration trend was very similar to those for other Ottawa sand columns except that the residual TCE after the air flushing showed relatively high. Column tests with different water contents were performed and results showed high removal efficiency even in a high water content sand column. Toluene as one of BTEX compounds was used in an Ottawa sand column and a real soil column. Removal trends were similar to those in TCE contaminated columns and more than 98% of initial toluene mass were removed with SVE in both column.

Impact of Application Rate of Non-ionic Surfactant Mixture on Initial Wetting and Water Movement in Root Media and Growth of Hot Pepper Plug Seedlings (비이온계 계면활성제 혼합물의 처리농도가 상토의 수분 보유 및 고추 플러그묘의 생장에 미치는 영향)

  • Choi, Jong-Myung;Moon, Byung-Woo
    • Horticultural Science & Technology
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    • v.29 no.1
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    • pp.16-22
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    • 2011
  • In developing soil wetting agent using polyoxyethylene nonylphenyl ether (PNE) and polyoxyethylene castor oil (1:1; v/v), the effect of application rates on changes in concentration of PNE, initial wetting of peatmoss + perlite (7:3) medium, and growth of hot pepper (Capsicum annuum L. 'Knockwang') plug seedlings were investigated. The elevation of application rates of wetting agent increased the amount of water retained by the root media. The treatment of 2.5 $mL{\cdot}L^{-1}$ showed similar water retention to + control ($AquaGro^L$ 3.0 $mL{\cdot}L^{-1}$). Most of the liquid wetting agent (LWA) incorporated during the medium formulation leached out in the first and second irrigation, then it decreased gradually until 10 times in irrigation. In investigation of the influence of LWA on position of water infiltrating into root media, the vertical water movements in treatments of 0.5, 1.0, and 1.5 $mL{\cdot}L^{-1}$ were much faster than those in 0.0 $mL{\cdot}L^{-1}$ (-control), but relative speed of water movement decreased by the elevation in application rate of LWA to 2.0 or 2.5 $mL{\cdot}L^{-1}$. The evaporative water loss of root media that to contained various rate of LWA and irrigated to reach container capacity was the fastest in -control among the treatments and it delayed as the application rate of LWA was elevated. The plant height of 22.2 cm in 0.5 $mL{\cdot}L^{-1}$ and stem diameter of 3.26 mm in 1.0 $mL{\cdot}L^{-1}$ were the highest among the treatments tested. The treatment of 1.0 $mL{\cdot}L^{-1}$ also had the heaviest fresh and dry weights such among treatments tested as 3.08 g and 0.861 g per plant, respectively. The elevated application rate over than 1.5 $mL{\cdot}L^{-1}$ resulted in decreased seedling growth. The results mentioned above indicate that optimum application rate of LWA is 1.0 $mL{\cdot}L^{-1}$.

EVALUATION OF PERIODONTAL LIGAMENT CELL VIABILITY IN RAT TEETH AFTER FROZEN PRESERVATION USING IN-VIVO MTT ASSAY (급속냉동된 쥐 치아의 in vivo MTT 검색법을 이용한 치주인대세포 활성도 평가)

  • Kim, Jae-Wook;Kim, Eui-Sung;Kim, Jin;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.31 no.3
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    • pp.192-202
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    • 2006
  • The purpose of this study was to examine the viability of PDL cells in rat molars by using in vivo MTT assay, which was used to compare fast cryopreservation group by liquid nitrogen $(-196^{\circ}C)\;with\;4^{\circ}C$ cold preservation group. A total of 74 Sprague-Dawley white female rats of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted as atraumatically as possible under ketamine anesthesia. Ten teeth of each group were divided as six experimental groups depending upon the preservation. Cryopreservation groups were Group 1 (5% DMSO 6% HES in F medium) Group 2 (10% DMSO in F medium), Group 3 (5% DMSO 6% HES in $Viaspan^(R)$). Group 4 (10% DMSO in $Viaspan^(R)$) which were cryopreserved for 1 week and cold preservation groups were Group 5 (F medium) , Group 6 ($Viaspan^(R)$) at $4^{\circ}C$ for 1 week. Immediate extraction group was used as a control. After preservation and thawing, the in vivo MTT assay was processed. Two way ANOVA and Duncan's Multiple Range Test was performed at the 95 % level of confidence, Another 2 teeth of each group were treated as the same manner and frozen sections $10{\mu}m$ thick for microscopic observation. The value of optical density obtained after in vivo MTT analysis was divided by the value of eosin staining for tissue volume standardization. Group 1, 2 had significantly higher optical density than Group 3 and 4 which had the lowest OD value. Group 6 had higher OD value than in Group 5 (P<0.05). Histological findings of periodontal ligament cell, after being stained with MTT solution were consistent with the in vivo MTT assay results. In this study, the groups which were frozen with DMSO as a cryoprotectant and the groups with F medium showed the best results.