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Development and Validation of the Analytical Method for Oxytetracycline in Agricultural Products using QuEChERS and LC-MS/MS (QuEChERS법 및 LC-MS/MS를 이용한 농산물 중 Oxytetracycline의 잔류시험법 개발 및 검증)

  • Cho, Sung Min;Do, Jung-Ah;Lee, Han Sol;Park, Ji-Su;Shin, Hye-Sun;Jang, Dong Eun;Cho, Myong-Shik;Jung, ong-hyun;Lee, Kangbong
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.227-234
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    • 2019
  • An analytical method was developed for the determination of oxytetracycline in agricultural products using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After the samples were extracted with methanol, the extracts were adjusted to pH 4 by formic acid and sodium chloride was added to remove water. Dispersive solid phase extraction (d-SPE) cleanup was carried out using $MgSO_4$ (anhydrous magnesium sulfate), PSA (primary secondary amine), $C_{18}$ (octadecyl) and GCB (graphitized carbon black). The analytes were quantified and confirmed with LC-MS/MS using ESI (electrospray ionization) in positive ion MRM (multiple reaction monitoring) mode. The matrix-matched calibration curves were constructed using six levels ($0.001{\sim}0.25{\mu}g/mL$) and coefficient of determination ($r^2$) was above 0.99. Recovery results at three concentrations (LOQ, $10{\times}LOQ$, and $50{\times}LOQ$, n=5) were from 80.0 to 108.2% with relative standard deviations (RSDs) less than of 11.4%. For inter-laboratory validation, the average recovery was in the range of 83.5~103.2% and the coefficient of variation (CV) was below 14.1%. All results satisfied the criteria ranges requested in the Codex guidelines (CAC/GL 40-1993, 2003) and the Food Safety Evaluation Department guidelines (2016). The proposed analytical method was accurate, effective and sensitive for oxytetracycline determination in agricultural commodities. This study could be useful for safety management of oxytetracycline residues in agricultural products.

The Usefulness of PCR Study in AFB Smear Negative Patients on Admission (내원시 항산균도말검사상 음성인 환자에서 실시한 PCR검사방법의 유용성에 대한 연구)

  • Kim, C.S.;Son, H.D.;Park, M.R.;Seo, J.Y.;Cho, M.D.;Rheu, N.S.
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.5
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    • pp.1001-1010
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    • 1997
  • Background : PCR technique is useful in diagnosis of pulmonary tuberculosis. But, its sensitivity and specificity is some different among several studies. Our aim is compare our PCR results with other's previous PCR results in AFB smear negative patients. Methods : PCR were performed in patients that their disease were suspected as active pulmonary tuberculosis and that their initial serial sputum AFB smear results were negative. Total number of patients studied by PCR technique was 177. Also, we analyzed the data only in patients whose bronchial washing fluid AFB smear was negative. And the primer had been used was IS 6110. Results : In our retrograde study, the number of patients who are diagnosed as having active pulmonary tuberculosis, inactive pulmonary tuberculosis and nontuberculous pulmonary disease was 99, 28, 50, respectively. In the sputum study, the sensitivity of PCR is 41.5% (27 PCR positive cases/65 active TBc cases). And the sensitivity of TB culture is 53.8% (35 TB culture positive cases/65 active TBc cases). In the bronchial washing specimen study, the sensitivity of PCR is 53.8% (21 PCR positive cases/39 active TBc cases). And the sensitivity of TB culture is 43.6% (17 TB culture Positive cases/39 active TBc cases). The specificity of PCR in our study is 94.9%. (74 PCR negative cases/78 inactive TBc or nontubereulous cases) In the cases of patients who were never takened anti-TBc medication, the sensitivity of PCR (45.6%--25 positive cases/55 cases) is some lower than culture (58.2%--32 positive cases/55 cases). In the cases of patients who had been takened anti-TBC medication. the sensitivity of PCR (60%--18 positive cases/30 cases) is some superior than culture (50%--15 positive cases/30 cases). Conclusion : We think that PCR results in cases of sputum AFB smear negative patients is nearly same as culture. And PCR is especially useful in patients who had been takened anti-TBc medication on admission.

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Intramuscular Administration of Zinc Metallothionein to Preslaughter Stressed Pigs Improves Anti-oxidative Status and Pork Quality

  • Li, L.L.;Hou, Z.P.;Yin, Y.L.;Liu, Y.H.;Hou, D.X.;Zhang, B.;Wu, G.Y.;Kim, S.W.;Fan, M.Z.;Yang, C.B.;Kong, X.F.;Tang, Z.R.;Peng, H.Z.;Deng, D.;Deng, Z.Y.;Xie, M.Y.;Xiong, H.;Kang, P.;Wang, S.X.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.761-767
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    • 2007
  • This study was conducted to determine the effects of exogenous zinc-metallothionein (Zn-MT) on anti-oxidative function and pork quality. After feeding a corn-soybean meal-based diet for two weeks, 48 pigs ($Duroc{\times}Landrace{\times}Chinese\;Black Pig$) were assigned randomly to four groups. Pigs in Group 1 were maintained under non-stress conditions, whereas pigs in Groups 2, 3 and 4 were aggressively handled for 25 min to produce stress. Pigs in Groups 1, 2, 3, and 4 received intramuscular administration of saline (control group; CON), 0 (negative control group; NCON), 0.8 (low dose group; LOW), and 1.6 (high dose group; HIGH) mg rabbit liver Zn-MT per kg body weight, respectively. Pigs were slaughtered at 3 and 6 h post-injection. Zn-MT treatment increased (p<0.05) the activities of superoxide dismutase (SOD) and glutathione-peroxidase (GSH-PX) while decreasing the concentration of malondialdehyde (MDA) in liver. These responses were greater (p<0.05) at 6 h than at 3 h post Zn-MT injection. Zn-MT treatment increased (p<0.05) hepatic SOD mRNA levels in a time and dose-dependent manner and decreased (p<0.05) serum glutamate-pyruvate transaminase and lactate dehydrogenase activities (indicators of tissue integrity). Zn-MT administration decreased (p<0.05) lactate concentration and increased (p<0.05) pH and water-holding capacity in the longissimus thorasis meat. Collectively, our results indicate that intramuscular administration of Zn-MT to pre-slaughter stressed pigs improved tissue anti-oxidative ability and meat quality.

Physicochemical Changes of Food Waste Slurry Co-fermented with Pig Manure Slurry (음식물쓰레기와 돈분 액상물의 혼합부숙시 이화학적 특성 변화)

  • So, Kyu-Ho;Seong, Ki-Seog;Hong, Seung-Gil
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.4
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    • pp.242-248
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    • 2007
  • To find a feasibility of utilization of food waste slurry (FWS) generated during composting, FWS was combined with pig manure slurry (PMS) in various ratios and the change of nutrient contents and offensive odor of the combined slurries before and after fermentation were studied. The initial pH was 7.67 for PMS and 8.45 for FWS. However, during the fermentation, pH increased in the combined slurries with the higher FWS rate among the treatments while decreased in thosewith higher PMS rate. EC of each slurry sample showed that the difference among combined slurry samples has been reduced during fermentation and became stabilized in $21{\sim}23dS\;m^{-1}$ after 180 days. After 180 days fermentation, total nitrogen (T-N) decreased. T-N of mixture with a half and more FWS decreased up to 0.1%, less than the critical level (0.3%). The contents of O.M., T-N, phosphorus, calcium and magnesium decreased with fermentation while those of potash and salinity increased. From initial fermentation until 30 days, a lot of $NH_3$, as an offensive odor, was produced. However, it decreased steadily, except in higher PMS rate. In terms of producing $50{\mu}g\;ml^{-1}$ of $NH_3$, the top layer took 30 days after fertilization with FWS only, 45 days for utilized treatment with F75 (25 % of PMS), 75 days for utilized with F50 (50%) and F25 (75%) and 90 days for PMS only, respectively. $RNH_2$ also had similar trend with $NH_3$ but it was produced continuously as long fermentation proceeded. In terms of $RNH_2$, the decrease in concentration up to $50{\mu}g\;ml^{-1}$ were; 45 days for FWS only(F100), 105 days for F75 utilization, 120 daysfor F50, 165 days for F25, respectively. ethyl mercaptan was produced in PMS until 180 days after fertilization but it was not produced in FWS. Sensory tests as an integrated test of offensive odor were also done. FWS showed lower than 1 after 30 days from initial fermentation, while PMS had still offensive odor even up to 180 days from initial fermentation. It is probably affected by the continuous production of ethyl mercaptan and amines. However, considering in decrease T-N content caused by volatilization while offensive odor intensity according to official standard of fertilizer is lower than 2. Further study on controlling offensive odor needs to be done.

The Clinical Study of the Lower Respiratory Tract Infection by Respiratory Syncytial Virus on Children under 2 Year of Age (2세 이하 어린이에서 Respiratory Syncytial Virus에 의한 하기도 감염에 관한 연구)

  • Cho, Jung Ik;Choi, Hyung Chul;Kim, Jong Duck;Cho, Ji Hyun
    • Pediatric Infection and Vaccine
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    • v.7 no.2
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    • pp.193-200
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    • 2000
  • Purpose : This study was designed and performed for evaluations of clinical manifestation and course of the children under 2 year of age with respiratory tract infection and positive respiratory syncytial virus(RSV) antigen. Methods : The selection criteria of the patients were children under 24 month-of-age, Clinical manifestation of respiratory tract infection, and positive RSV antigen that was detected by Vitek ImmunoDiagnostic Assay System(VIDAS) from nasal cavity. The additional laboratory and simple chest X-ray findings were reviewed from the chart of children who were admitted Wonkwang university hospital from October 1999 to March 2000. Results : Total number of patients enrolled on this study was 102. The 48(47%) children were RSV antigen positive by VIDAS method. Abnormal chest X-ray findings were noticed in 38 cases. The male to female sex ratio of 48 RSV antigen positive cases was 1.2 : 1. The mean and range of age was $10.2{\pm}5.9$ and 1.0~24 months. The peak outbreak of cases was noticed on November, 1999. All of the cases shows coughing but rale was audible in 30 cases(60%). Dyspnea, wheezing, and intercostal retraction were noticed 11(23%), 15(31%), and 10(21%) cases respectively. The most common chest X-ray finding was scattered patch infiltration that was noticed in 30 cases(63%). The mean total white blood cell counts in peripheral blood was $12,608{\pm}4,686/mm^3$. The mean blood level of IgA and IgE were $50.8{\pm}20.9$ and $72.1{\pm}98.3mg/dL$ respectively. The C-reactive protein was $16.0{\pm}18.5mg/L$. Total 5 cases need a mechanical respiraton. The duration of admission was under 7 days in 36 cases(75%). Conclusion : The RSV antigen was detected commonly in late fall and winter season. The severity of children under 2 years old with RSV respiratory tract infection take in some degree a gave courses.

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PROSPECTIVE CLINICAL EVALUATION OF THREE DIFFERENT BONDING SYSTEMS IN CLASS V RESIN RESTORATIONS WITH OR WITHOUT MECHANICAL RETENTION (접착제와 와동형성의 차이에 따른 5급 복합레진 수복의 전향적 임상연구)

  • Lee Kyung-Wook;Choung Sae-Joon;Han Young-Chul;Son Ho-Hyun;Um Chung-Moon;Oh Myoung-Hwan;Cho Byeong-Hoon
    • Restorative Dentistry and Endodontics
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    • v.31 no.4
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    • pp.300-311
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    • 2006
  • The purpose of this study is to evaluate prospectively the effect of different bonding systems and retention grooves on the clinical performance of resin restorations in non-carious cervical lesions (NCCLs). Thirty-nine healthy adults who had at least 2 NCCLs in their premolar areas were included in this study. One hundred and fifty teeth were equally assigned to six groups: (A) Scotchbond Multi-Purpose (SBMP, 3M ESPE, St. Paul, MN, USA, 4th generation bonding system) without retention grooves; (B) SBMP with retention grooves; (C) BC Plus (Vericom Co., Anyang, Gyeonggido, Korea, 5th generation bonding system) without retention grooves; (D) BC Plus with retention grooves; (E) Adper Prompt (3M ESPE, Seefeld, Germany, 6th generation bonding system) without retention grooves; (F) Adper Prompt with retention grooves. All cavities were filled with a hybrid composite resin. Denfil (Vericom Co., Anyang, Gyeonggido, Korea) by one operator. Restorations were evaluated at baseline and at 6-month recall, according to the modified USPHS (United States Public Health Service) criteria. Additionally, clinical photographs were taken and epoxy resin replicas were made for SEM evaluation. At 6-month recall, there were some differences in the number of alpha ratings among the experimental groups. But, despite the differences in the number of alpha ratings, there was no significant difference among the 3 adhesive systems (p < 0.05). There was also no significant difference between the groups with or without mechanical retention (p < 0.05). Follow-ups for longer periods than 6 months are needed to verify the clinical performance of different bonding systems and retention grooves.

Thought Experiments: on the Working Imagination and its Limitation (사고실험 - 상상의 작용과 한도에 대해)

  • Hwang, Hee-sook
    • Journal of Korean Philosophical Society
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    • v.146
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    • pp.307-328
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    • 2018
  • The use of thought experiments has a long history in many disciplines including science. In the field of philosophy, thought experiments have frequently appeared in the pre-existing literature on the contemporary Analytic Philosophy. A thought experiment refers to a synthetic environment where the designer of the experiment-with his or her intuition and imagination-tests common-sense knowledge. It can be understood as a conceptual tool for testing the validity of the common understanding of an issue or a phenomenon. However, we are not certain about the usefulness or efficacy of a thought experiment in knowledge production. The design of a thought experiment is meant to lure readers into believing as intended by the experiment itself. Thus, regardless of the purpose of a thought experiment, many readers who encounter the experiment could feel deceived. In this paper, to analyze the logic of thought experiments and to seek the source of uneasiness the readers and critics may feel about thought experiments, I draw lessons from three renowned thought-experiments: Thomson's 'ailing violinist', Putnam's 'brain in a vat', and Searle's 'Chinese room'. Imaginative thought experiments are usually constructed around a gap between the reality and the knowledge/information at hand. From the three experiments, several lessons can be learned. First, the evidence of the existence of a gap provided via thought experiments can serve as arguments for counterfactual situations. At the same time, the credibility and efficacy of the thought experiments can be damaged as soon as the thought-experiments are carried out with inappropriate and/or murky directions regarding the procedures of the experiment or the background of the study. According to D. R. Hofstadter and D. C. Dennett(1981), the 'knob setting' in a thought experiment can be altered in the middle of a simulation of the experimental condition, and then the implications of the thought experiment change altogether, indicating that an entirely different conclusion can be deduced from thought experiment. Lastly, some pre-suppositions and bias of the experiment designers play a considerable role in the validity and the chances of success of a thought experiment; thus, it is recommended that the experiment-designers refrain from exercising too much of their imagination in order to avoid contaminating the design of the experiment and/or wrongly accepting preconceived/misguided conclusions.

Cardio-pulmonary Adaptation to Physical Training (운동훈련(運動訓練)에 대(對)한 심폐기능(心肺機能)의 적응(適應)에 관(關)한 연구(硏究))

  • Cho, Kang-Ha
    • The Korean Journal of Physiology
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    • v.1 no.1
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    • pp.103-120
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    • 1967
  • As pointed out by many previous investigators, the cardio-pulmonary system of well trained athletes is so adapted that they can perform a given physical exercise more efficiently as compared to non-trained persons. However, the time course of the development of these cardio-pulmonary adaptations has not been extensively studied in the past. Although the development of these training effects is undoubtedly related to the magnitude of an exercise load which is repeatedly given, it would be practical if one could maintain a good physical fitness with a minimal daily exercise. Hence, the present investigation was undertaken to study the time course of the development of cardio-pulmonary adaptations while a group of non-athletes was subjected to a daily 6 to 10 minutes running exercise for a period of 4 weeks. Six healthy male medical students (22 to 24 years old) were randomly selected as experimental subjects, and were equally divided into two groups (A and B). Both groups were subjected to the same daily running exercise (approximately 1,000 kg-m). 6 days a week for 4 weeks, but the rate of exercise was such that the group A ran on treadmill with 8.6% grade for 10 min daily at a speed of 127 m/min while the group B ran for 6 min at a speed of 200 m/min. In order to assess the effects of these physical trainings on the cardio-pulmonary system, the minute volume, the $O_2$ consumption, the $CO_2$ output and the heart rate were determined weekly while the subject was engaged in a given running exercise on treadmill (8.6% grade and 127 m/min) for a period of 5 min. In addition, the arterial blood pressure, the cardiac output, the acid-base state of arterial blood and the gas composition of arterial blood were also determined every other week in 4 subjects (2 from each group) while they were engaged in exercise on a bicycle ergometer at a rate of approximately 900 kg m/min until exhaustion. The maximal work capacity was also determined by asking the subject to engage in exercise on treadmill and ergometer until exhaustion. For the measurement of minute volume, the expired gas was collected in a Douglas bag. The $O_2$ consumption and the $CO_2$ output were subsequently computed by analysing the expired gas with a Scholander micro gas analyzer. The heart rate was calculated from the R-R interval of ECG tracings recorded by an Offner RS Dynograph. A 19 gauge Cournand needle was inserted into a brachial artery, through which arterial blood samples were taken. A Statham $P_{23}AA$ pressure transducer and a PR-7 Research Recorder were used for recording instantaneous arterial pressure. The cardiac output was measured by indicator (Cardiogreen) dilution method. The results may be summarized as follows: (1) The maximal running time on treadmill increased linearly during the 4 week training period at the end of which it increased by 2.8 to 4.6 times. In general, an increase in the maximal running time was greater when the speed was fixed at a level at which the subject was trained. The mammal exercise time on bicycle ergometer also increased linearly during the training period. (2) In carrying out a given running exercise on treadmill (8.6%grade, 127 m/min), the following changes in cardio·pulmonary functions were observed during the training period: (a) The minute volume as well as the $O_2$ consumption during steady state exercise tended to decrease progressively and showed significant reductions after 3 weeks of training. (b) The $CO_2$ production during steady state exercise showed a significant reduction within 1 week of training. (c) The heart rate during steady state exercise tended to decrease progressively and showed a significant reduction after 2 weeks of training. The reduction of heart rate following a given exercise tended to become faster by training and showed a significant change after 3 weeks. Although the resting heart rate also tended to decrease by training, no significant change was observed. (3) In rallying out a given exercise (900 kg-m/min) on a bicycle ergometer, the following change in cardio-vascular functions were observed during the training period: (3) The systolic blood pressure during steady state exercise was not affected while the diastolic blood Pressure was significantly lowered after 4 weeks of training. The resting diastolic pressure was also significantly lowered by the end of 4 weeks. (b) The cardiac output and the stroke volume during steady state exercise increased maximally within 2 weeks of training. However, the resting cardiac output was not altered while the resting stroke volume tended to increase somewhat by training. (c) The total peripheral resistance during steady state exercise was greatly lowered within 2 weeks of training. The mean circulation time during exorcise was also considerably shortened while the left heart work output during exercise increased significantly within 2 weeks. However, these functions_at rest were not altered by training. (d) Although both pH, $P_{co2}\;and\;(HCO_3-)$ of arterial plasma decreased during exercise, the magnitude of reductions became less by training. On the other hand, the $O_2$ content of arterial blood decreased during exercise before training while it tended to increase slightly after training. There was no significant alteration in these values at rest. These results indicate that cardio-pulmonary adaptations to physical training can be acquired by subjecting non-athletes to brief daily exercise routine for certain period of time. Although the time of appearance of various adaptive phenomena is not identical, it may be stated that one has to engage in daily exercise routine for at least 2 weeks for the development of significant adaptive changes.

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Changes of Weed Community in Lowland Rice Field in Korea (한국(韓國)의 논 잡초분포(雜草分布) 현황(現況))

  • Park, K.H.;Oh, Y.J.;Ku, Y.C.;Kim, H.D.;Sa, J.K.;Park, J.S.;Kim, H.H.;Kwon, S.J.;Shin, H.R.;Kim, S.J.;Lee, B.J.;Ko, M.S.
    • Korean Journal of Weed Science
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    • v.15 no.4
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    • pp.254-261
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    • 1995
  • The nationwide weed survey was conducted in lowland rice fields over whole country of Korea in 1992 in order to determine a change of weed community and to identify a major dominant weed species and/or problem weed. Based on morphological characteristics of weeds, population ratio of broad leaf weed was 42.6%, grasses weed-9.0%, sedges-33.4% and others were 15.0%. Annual weed was 33.4% while perennial weed was 66.6% in terms of life cycle of weeds. Meanwhile, there was different weed occurrence as affected by planting method of the rice plant. In hand transplanted paddy fields predominant weed species was Sagittaria trifolia L., Monochoria vaginalis Presl., and Aneilema japonica Kunth. In machine transplanted rice fields of infant and young rice seedling Eleocharis kuroguwai Ohwi. and S. trifolia L. were more predominant. There was high occurrence of M. vaginalis, Echinochloa crus-galli L., and Leesia japonica Makino in water seeding while E. crus-galli and Cyperus serotinus Rottb. were predominant weed species in dry seeded rice. Monoculture of the rice plant would cause to high occurrence of E. kuroguwai, S. trifolia, M. vaginalis, E. crus-galli, and Sagittaria pygmaea Miq and there was higher population of S. trifolia, S. pygmaea, M. vaginalis, E crus-galli, and E. kuroguwai in double cropping system based on rice culture. In particular, there was high different weed occurrence under different transplanting times. E. kuroguwai, S. trifolia, S. pygmaea, M. vaginalis, and C. serotinus were higher population at the transplanting of 25 May and S. trifolia, E crus-galli, C. serotinus, and M. vaginalis at 10 June and S. pygmaea, E. kuroguwai, M. vaginalis, S. trifolia, and E. crusgalli at 25 June in Korea, respectively. Autumn tillage in terms of tillage time would cause more predominant weed species such as S. trifolia, E. kuroguwai, M. vaginalis, and S. pygmaea while spring tillage was higher population of E. kuroguwai, S. trifolia, E. crusgalli, M. vaginalis, and S. pygmaea. In plain area of paddy field there was higher occurrence of E. kuroguwai, S. trifolia, M. vaginalis, E. crus-galli, and S. pygmaea and in mid-mountainous area S. trifolia, E. kuroguwai, M. vaginalis, E. crus-galli, and Ludwigia prostrate Roxb. while in mountainous area S. trifolia, M. vaginalis, Potamogeton distinctus Ben., E. kuroguwai, and E. crus-galli were. In 1992 the most ten predominant weed species at the rice field of Korea based on summed dominant ratio(SDR) were E. kuroguwai > S. trifolia > E. crus-galli > M. vaginalis > S. pygmaea > C. serotinus > L. prostrate > P. distinctus > A. japonica > Scirpus juncoides Roxb.

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A Study on the Extraction of Monasil PCA using Liquid CO2 (액체 이산화탄소 이용한 Monasil PCA 추출에 대한 연구)

  • Cho, Dong Woo;Oh, Kyoung Shil;Bae, Won;Kim, Hwayong;Lee, Kab-Soo
    • Korean Chemical Engineering Research
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    • v.50 no.4
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    • pp.684-689
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    • 2012
  • Poly(acrylic acid) (PAA) microspheres is one of the widely-used polymeric materials for the bio-field application and the electric materials. For the synthesis of PAA microspheres, the polymerization technique using surfactants is applied. After the synthesis, the purification and separation processes are required for the removal of surfactant. When general organic solvents were used, many problems, such as huge amount of waste solvent, additional separation processes, and the possibility of residual media, were occurred. Thus, High-pressure Soxhlet extraction using liquid $CO_2$ was developed to solve these problems. In this study, High-pressure Soxhlet extraction of the synthesized PAA microspheres using liquid $CO_2$ was conducted for the removal of Monasil PCA which is used for the dispersion polymerization of acrylic acid in compressed liquid Dimethyl ether (DME). The morphology of the extracted PAA particles was checked by field emission scanning electron microscopy (FE-SEM) and the residual concentration of Monasil PCA was analyzed by inductively coupled plasma - Optical Emission Spectrometer (ICP-OES). For studying the effect of the solvent effect, Soxhlet extraction was conducted using n-hexane, liquid DME, and liquid $CO_2$. In case of n-hexane, some extracted PAA microspheres were produced. However, deformation was also occurred due to the high thermal energy of n-hexane vapor. Liquid DME could not remove Monasil PCA. When using liquid $CO_2$, the extracted PAA microspheres which were free for the residual solvent were produced without deformation. For finding the optimum operating condition, high-pressure Soxhlet extraction was conducted for 8 hours with changing the temperature of reboiler and condenser. When the extractor temperature is $19.6{\pm}0.2^{\circ}C$ and the pressure is $51.5{\pm}0.5$ bar, the best removal efficiency was obtained.