• Title/Summary/Keyword: system separation method

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Determination of methamphetamine, 4-hydroxymethamphetamine, amphetamine and 4-hydroxyamphetamine in urine using dilute-and-shoot liquid chromatography-tandem mass spectrometry (시료 희석 주입 LC-MS/MS를 이용한 소변 중 메스암페타민, 4-하이드록시메스암페타민, 암페타민 및 4-하이드록시암페타민 동시 분석)

  • Heo, Bo-Reum;Kwon, NamHee;Kim, Jin Young
    • Analytical Science and Technology
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    • v.31 no.4
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    • pp.161-170
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    • 2018
  • The epidemic of disorders associated with synthetic stimulants, such as methamphetamine (MA) and amphetamine (AP), is a health, social, legal, and financial problem. Owing to the high potential of their abuse and addiction, reliable analytical methods are required to detect and identify MA, AP, and their metabolites in biological samples. Thus, a dilute-and-shoot liquid chromatography-tandem mass spectrophotometry (LC-MS/MS) was developed for simultaneous determination of MA, 4-hydroxymethamphetamine (4HMA), AP, and 4-hydroxyamphetamine (4HA) in urine. Urine sample ($100{\mu}L$) was mixed with $50{\mu}L$ of mobile phase consisting of 0.4 % formic acid and methanol and $50{\mu}L$ of working internal-standard solution. Aliquots of $8{\mu}L$ diluted urine was injected into the LC-MS/MS system. For all analytes, chromatographic separation was performed using a C18 reversed-phase column with gradient elution and a total run time of 5 min. The identification and quantification were performed by multiple reaction monitoring (MRM). Linear least-squares regression was conducted to generate a calibration curve, with $1/x^2$ as the weighting factor. The linear ranges were 2.0-200, 1.0-800, and 10-2500 ng/mL for 4HA and 4HMA, AP, and MA, respectively. The inter- and intraday precisions were within 6.6 %, whereas the inter- and intraday accuracies ranged from -14.9 to 11.3 %. The low limits of quantification were 2.0 ng/mL (4HA and 4HMA), 1.0 ng/mL (AP), and 10 ng/mL (MA). The proposed method exhibited satisfactory selectivity, dilution integrity, matrix effect, and stability, which are required for validation. Moreover, the purification efficiency of high-speed centrifugation was clearly higher than 6-15 % for QC samples (n=5), which was higher than that of the membrane-filtration method. The applicability of the proposed method was tested by forensic analysis of urine samples from drug abusers.

Method Validation and Quantification of Lutein and Zeaxanthin from Green Leafy Vegetables using the UPLC System (UPLC를 이용한 lutein과 zeaxanthin의 분석법 검증 및 엽채류에서의 정량적 평가)

  • Kim, Suna;Kim, Ji-Sun
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.686-691
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    • 2012
  • The objective of this research is to present method development and validation for the simultaneous determination of lutein and zeaxanthin using ultra performance liquid chromatography (UPLC). Also, rapid quantification was performed on six green leafy vegetables (Allium tuberosum, Aster scaber, Hemerocallis fulva, Pimpinella brachycarpa, Sedum sarmentosum and Spinacia oleracea) that are commonly consumed in Korea. Separation and quantification were successfully achieved with a Waters Acquity BEH C18 ($50{\times}2.1mm$, $1.7{\mu}m$) column by 85% methanol within 5 min. Two compounds showed good linearity ($r^2$ > 0.9968) in $1-150{\mu}g/mL$. Limit of detection (LOD) and quantification (LOQ) for lutein and zeaxanthin were 1.7 and 5.1 g/mL and 2.1 and 6.3 g/mL, respectively. The RSD for intra- and inter-day precision of each compound was less than 10.69%. The recovery of each compound was in the range of 91.75-105.13%. Aster scaber and Spinacia oleracea contained significantly higher amounts of lutein ($4.06{\pm}0.24$ and $3.97{\pm}0.10mg$/100 g of fresh weight), respectively.

A Study of material analysis and its experimentation of metamorphosis and its utilities in Copper Alloy plates for contemporary metal craft (현대금속공예용 동합금판의 재료분석과 형질변환 실험 및 응용에 관한 연구)

  • Lim, Ock-Soo
    • Archives of design research
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    • v.17 no.4
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    • pp.241-250
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    • 2004
  • In this research, the copper alloy plates C2200, C5210, C7701, C8113 were selected to make datum and to identify further usage of metal craft experimentation. For its experimentation, the general welding and TIG welding methods were researched; for 2nd experimentation, the Reticulation and Electroforming skill's differences in color and temperature were researched. With these methods 3 different kinds of works are introduced for sample studies. For this research, Dr. Lee, Dong-Woo who works in Poongsan Metal Co, supported 4 kinds of copper alloy metals. Which are Commercial bronze (Cu-Zn), Deoxidiged Copper(Cu-Sn-P), Nickel Silver (Cu-Ni-Zn), and White Bronze (Cu-Ni); they were applied partly and wholly by the method of Laminatin, Reticulation, Fusing, and Electroforming skills. In case of C2200, the brass, the A. C. TIG welding method is better under 2mm slight plate; the D.C. TIG welding is better upper 2mm plate; and 250~300$^{\circ}C$ is recommended for remain heat treatment. In case of C5210, not having Hydrogen in high temperature return period, doesn't need Oxygen in high temperature and hardening in comparative high temperature neither, it is good for welding. It contains Sn 2-9% ad P 0.03-0.4% generally; and in accordance with the growth rate of Sn contain amount, the harden temperature boundary become broad. In case of cold moment after welding, they are recommended that higher speed TIG welding, smaller melting site and less than 200$^{\circ}C$ for pre-heating temperature. In case of C7701, the 10-20% Ni, 15-30% Zn are widely used.. If it is upper 30% Zn, it become (${\alpha}+{\beta}$) system and adhesive power rate become lower, and the productivity become lower in low temperature but the productivity become higher in high temperature. Nickel Silver's resistance of electricity is well; and the heatproof and incorrodibility is good, too. Lastly, in case of C8113, good at persistence in salty and grind; high in strength of high temperature. In case of white brass, contain 10-30% Nickel and hardened in high temperature and become single phrase. For these reason, the crystallization particles easily become large, if the resistance become higher small amount of Pb, P, S separation rate become higher.

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Liver Splitting Using 2 Points for Liver Graft Volumetry (간 이식편의 체적 예측을 위한 2점 이용 간 분리)

  • Seo, Jeong-Joo;Park, Jong-Won
    • The KIPS Transactions:PartB
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    • v.19B no.2
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    • pp.123-126
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    • 2012
  • This paper proposed a method to separate a liver into left and right liver lobes for simple and exact volumetry of the river graft at abdominal MDCT(Multi-Detector Computed Tomography) image before the living donor liver transplantation. A medical team can evaluate an accurate river graft with minimized interaction between the team and a system using this algorithm for ensuring donor's and recipient's safe. On the image of segmented liver, 2 points(PMHV: a point in Middle Hepatic Vein and PPV: a point at the beginning of right branch of Portal Vein) are selected to separate a liver into left and right liver lobes. Middle hepatic vein is automatically segmented using PMHV, and the cutting line is decided on the basis of segmented Middle Hepatic Vein. A liver is separated on connecting the cutting line and PPV. The volume and ratio of the river graft are estimated. The volume estimated using 2 points are compared with a manual volume that diagnostic radiologist processed and estimated and the weight measured during surgery to support proof of exact volume. The mean ${\pm}$ standard deviation of the differences between the actual weights and the estimated volumes was $162.38cm^3{\pm}124.39$ in the case of manual segmentation and $107.69cm^3{\pm}97.24$ in the case of 2 points method. The correlation coefficient between the actual weight and the manually estimated volume is 0.79, and the correlation coefficient between the actual weight and the volume estimated using 2 points is 0.87. After selection the 2 points, the time involved in separation a liver into left and right river lobe and volumetry of them is measured for confirmation that the algorithm can be used on real time during surgery. The mean ${\pm}$ standard deviation of the process time is $57.28sec{\pm}32.81$ per 1 data set ($149.17pages{\pm}55.92$).

Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Ultrasensitive Enzymeimmunoassay for Testosterone in Human Saliva (사람 타액내 Testosterone의 초감도 효소면역측정법)

  • 윤용달;전은현;이창주;도병록;이준영
    • Development and Reproduction
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    • v.4 no.1
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    • pp.115-123
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    • 2000
  • A few enzymeimmunoassay (EfA) for testosterone (T) have been reported but was not suitable for all biological samples. The present study was designed to develop a rapid, ultrasensitive EIA and to apply this technique for study the physiological changes of T in biological samples. Saliva samples were collected at 06:00~09:00 hour during one menstrual cycle from 18 normally menstruating women and on 09:00~10:00 hour from 20 normal men. The present study shows an established EIA for testosterone, using horseradish peroxidase (HRP), which was covalently bonded to testosterone-3-carboxymethyloxime (T-3-CMO). One batch of T-antisera was also covalently linked to microcrystalline cellulose particles by a mixed anhydride method in order to facilitate separation of bound and free steroids. The established EIA was validated in terms of sensitivity, accuracy, specificity, precisions etc., comparing with conventional radioimmunoassay. The sensitivity of the established EIA was less than 25 pg/tube. The correlation coefficients between the expected T-values and observed T-values measured by EIA or RIA were r=0.985 and r=0.941 respectively. The cross reactivity of antiserum in EIA was a little higher than that of RIA, especially by 5 ${\alpha}$-DHT. The intra- and inter-assay precisions of the present EIA were similar to those of RIA. The present study also demonstrates that the normal T-values in saliva of Korean male & female samples are 265.65${\pm}$15.80 pmol/l and 109.74${\pm}$ 12.01 pmol/l, respectively. The present EIA seems to be established and suitable for use in the endocrinological studies. The advantages of this EIA system also might make the present T-EIA an ideal procedure for use in a routine assay of ordinary laboratory with a conventional spectrophotometer.

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Practical Applications of DNA Marker-Assisted Selection and OPU-Derived IVF Embryo Transfer for the Production of High Quality Meat in Hanwoo II. Production of IVF Embryos Derived Transvaginal Ovum Pick-up from DNA Marker-Proved Hanwoo (유전자 분석을 통하여 선발된 한우로부터 초음파 유래 체외수정란 이식에 의한 고품질 한우 생산기술의 실용화 II. DNA 검정우로부터 초음파 유래 체외수정란의 생산에 관한 연구)

  • 박희성;이지삼;진동인;박준규;홍승표;이명열;정장용
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.193-201
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    • 2001
  • This study was designed to examine the factors affecting in fertilization and development of embryos in vitro, and to examine whether zone drilling by laser irradiation can improve the hatching rate of IVF embryos from DNA marker-proved Hanwoo. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local anesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analyzed using the GLM procedure of SAS. Sperm separation methods did not have any significant effect on cleavage or developmental abilities of IVF embryos. Significantly(P<0.05) higher cleavage rate was observed in embryos from GI(60.0%, 3/5), GII(69.2%, 18/26) and GIII(62.1%, 59/95) compared to embryos from GIV oocytes(36.2%, 25/69). And the developmental rate to blastocyst stage was higher(P<0.05) in embryos from GI(33.3%, 1/3) and GII oocytes(38.9%, 7/18) than those from GIII(16.9%,10/59) and GIV oocytes(4.0%, 1/25). There was no significant difference in development of IVF embryos to blastocyst by media for in vitro culture. Proportion of hatched blastocyst was significantly(P<0.05) higher in embryos received zona drilling by laser than those of non-drilled.

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Survey of Beet Red Contents in Foods using TLC, HPLC (TLC, HPLC를 이용한 식품 중 비트레드 함량조사)

  • Jang, Yaung-Mi;Lee, Tal-Soo;Hong, Ki-Hyoung;Park, Sung-Kwan;Park, Sung-Kug;Kwon, Yong-Kwan;Park, Jae-Seok;Chang, Sun-Young;Hwang, Hye-Shin;Kim, Eun-Jeong;Han, Yeun-Jeong;Kim, Byung-Sub;Won, Hye-Jin
    • Journal of Food Hygiene and Safety
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    • v.20 no.4
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    • pp.244-252
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    • 2005
  • This study was performed far development of new analytical method of beet red in foods. In this study, analysis of beet red in foods has been carried out by detection of betanine and isobetanine, the main color component of beet red as indicator compounds. The qualitative analysis technique consisted of clean-up of the colors with a $C_{18}$ cartridge, separation of the colors by cellulose TLC plate using acetone:3-methyl-1-butanol:distilled water (7:7:6) as a solvent system. Also, the quantitative analysis was performed using X-terra RP at wavelength 538 nm and $0.1\%$ phosphoric acid : methanol (90:10) as a solvent. The quantitative results of beet .ed were as follows:900.22$\∼$27701.60 $\mu$g/g for 60 item in nutrient supplement food, $21.95\∼713.40{\mu}g/g$ for 30 items and N.D. for 18 items in cindy, and $155.85{\∼}505.37{\mu}g/g$ for 12 items in ice creams, $43.52\∼64.75{\mu}g/g$ for 18 items and N.D. for 54 item in sauce, N.D. for 12 items in retort food.

Feasibility Study of Isodose Structure Based Field-in-Field Technique for Total Body Irradiation (전신조사방사선치료 시 Isodose Structure를 이용한 Field-in-Field Technique의 유용성 평가)

  • Lee, Yoon Hee;Ban, Tae Joon;Lee, Woo Seok;Kang, Tae Young;Back, Geum Mun
    • The Journal of Korean Society for Radiation Therapy
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    • v.25 no.1
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    • pp.15-24
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    • 2013
  • Purpose: In Asan Medical Center, Two parallel opposite beams are employed for total body irradiation. Patients are required to be in supine position where two arms are attached to mid axillary line. Normally, physical compensators are required to compensate the large dose difference for different parts of body due to the different thicknesses compared to the umbilicus separation. There was the maximum dose difference up to 30% in lung and chest wall compared to the prescription dose. In order to resolve the dose discrepancy occurring on different body regions, the feasibility of using Fieid-in-Field Technique is investigated in this study. Materials and Methods: CT scan was performed to The RANDO Phantom with fabricated two arms and sent to Eclipse treatment planning system (version 10.0, Varian, USA). Conventional plan with physical lead compensator and new plan using Field-in-Field Technique were established on TPS. AAA (Anisotropic Analytical Algorithm) dose calculation algorithm was employed for two parallel opposite beams attenuation. Results: The dose difference between two methods was compared with the prescription dose. The dose distribution of chest and anterior chest wall uncovered by patient arms was 114~124% for physical lead compensator while Field-in-Field Technique gave 106~107% of the dose distribution. In-vivo dosimetry result using TLD showed that the dose distribution to the same region was 110~117% for conventional physical compensator and 104~107% for Field-in-Field Technique. Conclusion: In this study, the feasibility of using FIF technique has been investigated with fabricated arms attached Rando phantom. The dose difference was up to 17% due to the attached arms. It is shown that the dose homogeneity is within ${\pm}10%$ with the CT based 3-dimensional 4 step FIF technique. The in-vivo dosimetry result using TLD was showed that 95~107% dose distribution compared to prescription dose. It is considered that CT based 3-dimensional Field-in-Field Technique for the total body irradiation gives much homogeneous dose distribution for different body parts than the conventional physical compensator method and might be useful to evaluate the dose on each part of patient body.

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Developmental Plan of Man-Overboard Alert Devices of Small Fishing Vessels: A Study (소형어선의 선외추락 경보장치 개발 방안 연구)

  • Kim, Jae-Won;Kim, Byung-Ok;Lim, Jung-Gyun;Lee, Ju-Han;Yim, Jea-Hong;Park, Dong-Kook
    • Journal of Navigation and Port Research
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    • v.42 no.4
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    • pp.245-252
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    • 2018
  • A method of transmitting an alert signal in case of man-overboard (MOB) systems in a small fishing vessel navigating within coastal area is being operated as VHF-DSC equipment via a distress alert button and V-P ass Equipment via alert button or beacon separation. However, a small fishing vessel with a couple of crews on board is an inappropriate way to alert a man-overboard condition. However, internationally, MOB equipment using VHF-DSC, AIS, and Bluetooth technologies is used to transmit alert signals directly to the mother ship and other radio stations. In order to analyze the performance and technology of the MOB equipment operating in foreign countries, it was confirmed that the alarm signal can be received within a maximum of one nautical mile when the MOB device is on the water surface. An MOB device that meets domestic conditions needs to send an alarm signal to a station within the VHF communication range. However, in order to reduce the false alert signal, it is most appropriate to operate the VHF-DSC radio equipment installed on the ship remotely. Analysis of various technologies connecting the MOB and the VHF-DSC revealed that the Bluetooth system has advantages such as device miniaturization. When an emergency signal is transmitted from the MOB device, it can be received by a dedicated receiver and recognized through an external input terminal of the VHF-DSC equipment generating its own alarm. If the emergency situation cannot be processed at the mother ship, a distress alert is sent to all radio stations via VHF-DSC for response under emergencies faced by small fishing vessels.