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Quantitative determination of inosine 5'-monophosphate dehydrogenase activity in human peripheral blood mononuclear cells by ion-pair reversed-phase high-performance liquid chromatography (이온쌍 역상 HPLC를 이용한 인체 말초혈액단핵구에서 이노신 5'-일인산 탈수소효소 활성의 정량적 측정)

  • Shin, Hye-Jin;Kwon, Soon-Ho;Park, Ji-Myeong;Kwon, Soon-Hyo;Lee, Kyoung-Ryul;Kim, Young-Jin;Lee, Sang-Hoo
    • Analytical Science and Technology
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    • v.23 no.6
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    • pp.531-536
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    • 2010
  • A quantitative analytical method has been established for the measurement of inosine 5'-monophosphate dehydrogenase (IMPDH) activity in human peripheral blood mononuclear cells (PBMCs) by ion-pair reversed-phase high performance liquid chromatography equipped with ultraviolet detection (HPLC/UV). IMPDH is a ${\beta}$-nicotinamide adenine dinucleotide hydrate (NAD+)-dependent dehydrogenase in which the enzyme converts inosine 5'-monophosphate (IMP) into xanthosine 5'-monophosphate (XMP). Its activity was measured by quantifying a HPLC chromatogram corresponding to XMP produced during the incubation of lysed PBMCs with IMP as a substrate and $NAD^+$ as a coenzyme. XMP produced was detected at a wavelength of 260 nm. The mobile phase was composed of a mixture of 37 mM potassium dihydrogen phosphate containing 7 mM tetra-n-butylammonium hydrogen sulfate adjusted to pH 5.5 and methanol (85:15, v/v) with a flow rate of 1 mL/min. The calibration curve was linear ($r^2$=0.999999) in the range of $0.2-50.0\;{\mu}M$ and the limit of quantification (LOQ) was $0.2\;{\mu}M$. The intra- and inter-day precisions were between 0.88-1.47% and 0.85-5.24%, respectively. The intra- and inter-day accuracies were between 98.74-99.99% and 99.95-101.65%, respectively. IMPDH activity in 11 Korean healthy volunteers ranged from 18.29 to 36.60 nmol/h/mg protein (mean = $27.70{\pm}6.28\;nmol/h/mg$ protein).

Development and validation of an analytical method for nematicide imicyafos determination in agricultural products by HPLC-UVD (HPLC-UVD를 이용한 살선충제 imicyafos의 시험법 개발 및 검증)

  • Do, Jung-Ah;Park, Hyejin;Kwon, Ji-Eun;Choi, Won-Jo;Lee, Hyun-Sook;Chang, Moon-Ik;Hong, Jin-Hwan;Oh, Jae-Ho
    • Analytical Science and Technology
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    • v.27 no.5
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    • pp.234-242
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    • 2014
  • Imicyafos which is a nematicide for controlling root-knot nematodes has been registered in the Republic of Korea in 2012, and the maximum residue limits of imicyafos are set to watermelon and korean melon as each 0.05 mg/kg. Extremely reliable and sensitive analytical method is required for ensuring food safety on imicyafos residues in agricultural commodities. Imicyafos residues in samples were extracted with acetone, partitioned with hexane and dichloromethane, and then purified with florisil. The purified samples were analyzed by HPLC-UVD and confirmed with LC-MS. Linear range was between 0.1~5 mg/kg with the correlation coefficient ($r^2$) 0.99997. Average recoveries of imicyafos ranged from 77.0 to 115.4% at the spiked levels of 0.02 and 0.05 mg/kg with the relative standard deviations of 2.2~9.6%. Limit of detection and quantification were 0.005 and 0.02 mg/kg, respectively. An inter-laboratory study was conducted to validate the determination method in depth, and the results were satisfactory. All of the validation results revealed that the developed analytical method in this study is relevant for imicyafos determination in agricultural commodities and will be used as an official analytical method.

Development of a Simultaneous Analytical Method for Determination of Trinexapac-ethyl and Trinexapac in Agricultural Products Using LC-MS/MS (LC-MS/MS를 이용한 농산물 중 식물생장조절제 Trinexapac-ethyl과 대사산물 Trinexapac의 동시분석법 개발)

  • Jang, Jin;Kim, Heejung;Ko, Ah-Young;Lee, Eun-Hyang;Ju, Yunji;Chang, Moon-Ik;Rhee, Gyu-Seek;Suh, Saejung
    • Korean Journal of Environmental Agriculture
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    • v.34 no.4
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    • pp.318-327
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    • 2015
  • BACKGROUND: Trinexapac-ethyl is a plant growth regulator (PGR) that inhibits the biosynthesis of plant growth hormone (gibberellin). It is used for the prevention of lodging, increasing yields of cereals, and reducing mowing of turf. The experiment was conducted to establish a determination method for trinexapac-ethyl and its metabolites trinexapac in agricultural products using LC-MS/MS.METHODS AND RESULTS: Trinexapac-ethyl and trinexapac were extracted from agricultural products with methanol/ distilled water and the extract was partitioned with dichloromethane and then detected by LC-MS/MS. Limit of detection(LOD) was 0.003 mg/kg and limit of quantification(LOQ) was 0.01 mg/kg, respectively. Matrix matched calibration curves were linear over the calibration ranges (0.01-1.0 mg/L) for all the analytes into blank extract withr2> 0.997. For validation purposes, recovery studies were carried out at three different concentration levels (LOQ, 10LOQ, 50LOQ,n=5). Recoveries of trinexapacethyl and trinexapac were within the range of 73.6-106.9%, 72.7-99.2%, respectively. The relative standard deviations (RSDs) were less than 9.0%. All values were consistent with the criteria ranges requested in the CODEX guideline(CAC/GL 40, 2003).CONCLUSION: The proposed analytical method was accurate, effective and sensitive for trinexapac-ethyl and trinexapac determination and it can be used to as an official method in Korea.

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.

Risk Assessment Aluminum Levels of Circulating Agricultural Products in Korea (국내 다소비 농산물의 알루미늄 농도 모니터링 및 농산물 섭취로 인한 위해도 평가)

  • An, Jae-Min;Hong, Kyong-Suk;Kim, Sung-Youn;Kim, Ji-Hye;Yu, Kyong-Eun;Kim, Hyo-Young;Park, Hyoung-Dal;Lee, Jae-Hwon;Kim, Dong-Ho
    • Korean Journal of Environmental Agriculture
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    • v.34 no.4
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    • pp.336-344
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    • 2015
  • BACKGROUND: This study was carried out to collect occurrence data on aluminum in 12 type agricultural products and assess dietary exposure risk to the Korean population health for aluminum concentration in agricultural products.METHODS AND RESULTS: Aluminum analysis samples were performed using microwave device and Inductively Coupled Plasma Optical Emission Spectrometer. The LOD(Limit of Detection) for aluminum was 0.851 μg/kg, while the LOQ(Limit of Quantitation) was 2.838 μg/kg and recovery was 97.6% for aluminum. The average levels of aluminum in mg/kg were 0.526 for rice, 0.546 for Korean cabbage, 1.316 for corn, 6.207 for soybean, 0.549 for sweet potato, 0.257 for potato, 6.963 for spinach, 1.213 for carrot, 0.524 for garlic, 0.950 for radish, 1.015 for leek, and 3.511 for Welsh onion. The dietary exposures of aluminum through usual intake were polished rice 89.31 μg/day, Korean cabbage 33.14 μg/day, corn 0.66 μg/day, soybean 3.72 μg/day, sweet potato 6.86 μg/day, potato 4.96 μg/day, spinach 45.96 μg/day, carrot 6.79 μg/day, garlic 2.36 μ g/day, radish 7.32 μg/day, leek 2.23 μg/day and Welsh onion 43.89 μg/day, taking 0.57%, 0.21%, 0.00%, 0.02%, 0.04%, 0.03%, 0.04%, 0.04%, 0.02%, 0.05%, 0.01% and 0.28% of PTWI(2 mg/kg b.w./week), respectively.CONCLUSION: The levels of overall dietary exposure to aluminum for Korean population through intake of agricultural product was far below the recommended JECFA level, indicating of least possibility of risk.

Detection of Metagonimus sp. (Family Heterophyidae) Metacercariae from Freshwater Fishes in the Vicinity of Tong River, Kangwon Province, Korea (동강유역 담수어에 기생하는 흡충(Metagonimus sp.)의 피낭유출 감염 조사)

  • 전계식;김태선
    • Korean Journal of Environmental Biology
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    • v.17 no.2
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    • pp.169-173
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    • 1999
  • The present study was undertaken to survey the infection status of Metagonimus sp. metacercariae in freshwater fishes from Tong river located in Kangwon Province, a total of 178 freshwater fishes of 11 species were collected by a fish net and traps from the end of September to early October 1998. They were brought to the laboratory and examined under a stereomicroscope after artificial digestion with pepsin-HCI solution for one hour. Ten out of eleven species of freshwater fishes examined were found to contain metacercariae of Metagonimus sp. The most frequently infected fish was Liobagrus andersoni Regan 91.7% (11 of 12 fishes), followed by Gobiobotia breviburba Mori 75.0% (6 of 8 fishes), Zacco temmincki 66.2% (49 of 74 fishes), Cobitis koreensis Kim 50.0% (1 of 2 fishes), Microphysogobio longidorsails Mori 42.9% (9 of 21 fishes), Cobitis rotundicudata Wakiya et Mori 33.3% (2 of 6 fishes), Hemibarbus longirostris 33.3% (1 of 3 fishes), Pungtungia tenuicorpus 30.0% (3 of 10 fishes), Coreoleuciscus splenddus Mori 28.6% (4 of 14 fishes), Coreoperca herri Herzeanstein 11.1% (1 of 9 fishes) respectively Metagonimus metacercariae was not found from Moroco kumgangensis Uchida of freshwater fish. The total number of Metagonimus sp. metacercariae was 81, 17, 216, 1, 14, 2, 1, 4, 10, 1, respectively. Infection rates of Metagonimus sp. in the five districts was shown as follows : Chongson-up Kyulam-ri 50.0% (12 of 24 fishes), Chongson-up Kasu-ri 34.0% (17 of 50 fishes), Shindong-up Unchi-ri 56.0% (14 of 25 fishes), Shingdong-up Kosong-ri 57.1% (20 of 35 fishes), Shingdong-up Duckchon-ri 54.5% (24 of 44 fishes) respectively. Infection rate of freshwater fish was 48.9% (87 of 178 fishes) and from the results obtained in this survey, it was confirmed that freshwater fishes from Tong river was infected with metacercariae of Metagonimus sp.

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Urban Climate Impact Assessment Reflecting Urban Planning Scenarios - Connecting Green Network Across the North and South in Seoul - (서울 도시계획 정책을 적용한 기후영향평가 - 남북녹지축 조성사업을 대상으로 -)

  • Kwon, Hyuk-Gi;Yang, Ho-Jin;Yi, Chaeyeon;Kim, Yeon-Hee;Choi, Young-Jean
    • Journal of Environmental Impact Assessment
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    • v.24 no.2
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    • pp.134-153
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    • 2015
  • When making urban planning, it is important to understand climate effect caused by urban structural changes. Seoul city applies UPIS(Urban Plan Information System) which provides information on urban planning scenario. Technology for analyzing climate effect resulted from urban planning needs to developed by linking urban planning scenario provided by UPIS and climate analysis model, CAS(Climate Analysis Seoul). CAS develops for analyzing urban climate conditions to provide realistic information considering local air temperature and wind flows. Quantitative analyses conducted by CAS for the production, transportation, and stagnation of cold air, wind flow and thermal conditions by incorporating GIS analysis on land cover and elevation and meteorological analysis from MetPhoMod(Meteorology and atmospheric Photochemistry Meso-scale model). In order to reflect land cover and elevation of the latest information, CAS used to highly accurate raster data (1m) sourced from LiDAR survey and KOMPSAT-2(KOrea Multi-Purpose SATellite) satellite image(4m). For more realistic representation of land surface characteristic, DSM(Digital Surface Model) and DTM(Digital Terrain Model) data used as an input data for CFD(Computational Fluid Dynamics) model. Eight inflow directions considered to investigate the change of flow pattern, wind speed according to reconstruction and change of thermal environment by connecting green area formation. Also, MetPhoMod in CAS data used to consider realistic weather condition. The result show that wind corridors change due to reconstruction. As a whole surface temperature around target area decreases due to connecting green area formation. CFD model coupled with CAS is possible to evaluate the wind corridor and heat environment before/after reconstruction and connecting green area formation. In This study, analysis of climate impact before and after created the green area, which is part of 'Connecting green network across the north and south in Seoul' plan, one of the '2020 Seoul master plan'.

Sex Detection and In Vitro Development of Biopsied Bovine Embryo for LAMP Based Embryo Sexing (LAMP 방법에 의한 소 수정란의 성 판별과 Biopsy에 따른 수정란의 체외발달)

  • Cho S. R.;Choi S. H.;Kim H. J.;Han M. H.;Choe C. Y.;Chung Y. G.;Son D. S.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.169-176
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    • 2005
  • Loop-mediated isothermal amplification (LAMP) is novel DNA amplification methods that amplifies a target sequence specifically under isothemal condition. The present study was to assess the in vitro viability afier biopsy and sexing rate of different types of embryo biopsied. In vivo compact morulae and blastocyst embryos were obtained from Korean Native Cow (KNC) superovulated with FSH (Antorin, R-10) on 7 Day after artificial insemination. in vitro compact morulae and blastocyst embryos were obtained with KNC or Holsteins that were gained on 6, 7 or 8 day after in vitro fertilization(IVF) with frozen semen. Biopsy of bovine embryo was carried out in a $80{\mu}l$ drop with $Ca^{2+}-Mg^{2+}$ free D-PBS and the viability of biopsied embryos were evaluated in IVMD (IFP, Japan) medium at 12 hrs culture time. The sex ratio of biopsied Hanwoo embryos were male vs. female of $43.5\%\;vs.\;56.5\%$ in vivo and $33.9\%\;vs.\;49.2\%$ in vitro respectively, and male rate of biopsied Holstein embryos were significantly higher than female $(70.8\;vs.\;29.2\%)$. and indefinite rate of in vitro embryos was $16.9\%$ and in vivo was not. The degeneration rate of biopsied embryo, in vitro embryos were significantly higher than in vivo $(13.2\%\;vs,\;0.0\%,\;p<0.05)$. The survivability of in vivo embryo were between biopsied following punching method was significantly (P<0.05) higher than bisection method produced embryos $(100\%\;vs.\;83.3\%)$ and in vitro had no difference. However, the degeneration rate of biopsied embryo by bisection method was significantly higher than punching methods between in vivo and in vitro $(16.7\;vs.\;22.6\%,\;respectively,\;p<0.05)$. In conclusion, these results indicate that punching method was optimal and survivability after embryo biopsy was useful for reducing the damage caused by the embryo biopsy procedure for LAMP-based embryo sexing.

Embryo Production in Superior Hanwoo Donors and Embryo Transfer (우수 한우의 수정란 생산 및 이식)

  • Son D.S.;Han M.H.;Choe C.Y.;Choi S.H.;Cho S.R.;Kim H.J.;Ryu I.S.;Choi S.B.;Lee S.S.;Kim Y.K.;Kim S.K.;Kim S.H.;Shin K.H.
    • Journal of Embryo Transfer
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    • v.21 no.2
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    • pp.147-156
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    • 2006
  • The objective of this study was to supply excellent genetic resources to livestock farms by transferring embryos produced by genetically superior Korean cows (Hanwoo). Eighty Hanwoo donors were superovulated with gonadotropin ($Folltrpin^(R)\;or\;Antorin^(R)$) for 4 days combined with or without progesterone releasing intravaginal device (CIDR) insertion. The collected fresh or frozen-thawed embryos were transferred to 226 farm recipients. In this study, the effect of CIDR insertion in combination with gonadotropin ($Folltrpin^(R)$) treatments initiated at the random stage of estrous cycle on embryo production was evaluated and compared to conventional superovulation protocol. Moreover, the effect of gonadotropin ($Antorin^(R)$) dose in CIDR-treated Hanwoo donors on the embryo yield was determined. In addition, the effects of embryos (fresh vs. frozen-thawed), embryo transfer person, seasons and farms on the pregnancy rate were evaluated. In Hanwoo donors, CIDR insertion in combination with $Folltrpin^(R)$ treatments regardless of estrous detection resulted in increased numbers of total ova (6.5 vs. 5.8) and transferable embryos (3.9 vs. 3.2) compared to the conventional superovulation protocol (p<0.01). In CIDR-treated Hanwoo donors, the higher dose of $Antorin^(R)$ (36 vs. 28 mg) resulted in the increased number of transferable embryos (8.3 vs. 5.4, p<0.05). The embryos (fresh 43.9% vs. frozen-thawed 23.1%) and embryo transfer person (53.9 vs. $0{\sim}16.7%$) significantly affected the pregnancy rate after embryo transfer (p<0.01). These results suggest that CIDR-based superovulation protocol may be effectively used for production of superior Hanwoo embryos and, multiple ovulation and embryo transfer in Hanwoo might be effectively applied for livestock improvement if pregmancy rate with frozen-thawed embryos and embryo transfer skill would be improved.

In Vivo Embryo Production and Embryo Transfer in Hanwoo and Jeju Black Cattle Using CIDR (CIDR를 이용한 제주 한우 및 흑우의 체내 수정란 생산과 이식)

  • Kim, Y.H.;Koo, J.C.;Oh, C.W.;Kang, S.Y.;Yang, B.S.;Oh, S.J.;Kim, C.N.;Song, J.Y.;Kim, I.H.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.191-198
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    • 2006
  • The objectives of this study was to produce valuable offsprings of Hanwoo and Jeju black cattle using in vivo embryo production and embryo transfer techniques during 5 years ($2001{\sim}2005$) in Jeju. Two hundred and eighty six Hanwoo and sixty nine Jeju black cattles, at random stages of the estrous cycle, received a CIDR. Seven days later, the animals were superovulated with a total of 400 mg pFSH ($Folltropin^{(R)}-V$) administered twice daily in constant doses (each 50 mg) over 4 days. On the 6th administration of FSH, CIDR was withdrawn and 25 mg $PGF_{2{\alpha}}$ was administered. Cows were artificially inseminated thrice after estrous detection at 12 hr intervals. The cows received $250{\mu}g$ GnRH at the time of 2nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Recipients were synchronized with donors by insertion of a CIDR for 7 days and administration of 25 mg $PGF_{2{\alpha}}$ at the time of CIDR removal. The collected embryos were transferred to 1,219 recipients by 6 transfer persons. The mean numbers of total ova and transferable embryos from Hanwoo and Jeju black cattle donors were 7.4 and 4.7, respectively The number of transferable embryos differed between Hanwoo (5.0) and Jeju black cattle (3.5, p<0.05), while that of total ova did not differ. Repeated superovulation treatments decreased (p<0.05) the ratio of numbers of the flushed animals vs. superovulated animals in Jeju black cattle, and the numbers of total ova and transferable embryos in Hanwoo. More transferable embryos were collected at summer (5.6) than winter (2.9, p<0.01). The mean pregnancy rate was 40%. The pregnancy.ate was affected by transfer year (2001<2004, p<0.05) and transfer person ($33.0{\sim}41.9%$, p<0.01), while not by donor (embryo) breed. These results showed that in vivo embryo preduction and embryo transfer techniques using CIDR for Hanwoo and Jeju black cattle donors as well as recipient, regardless of their estrous cycle, may enable a stable embryo production and recipient preparation.