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Development and Validation of HPLC Method for Pharmacokinetic Study of Promethazine in Human (염산프로메타진 체내동태 연구를 위한 혈청 중 프로메타진의 HPLC 정량법 개발 및 검증)

  • Cho, Hae-Young;Kang, Hyun-Ah;Lee, Hwa-Jeong;Choi, Hoo-Kyun;Lee, Yong-Bok
    • Journal of Pharmaceutical Investigation
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    • v.36 no.1
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    • pp.23-29
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    • 2006
  • A rapid, selective and sensitive reversed-phase HPLC method for the determination of promethazine in human serum was developed, validated, and applied to the pharmacokinetic study of promethazine. Promethazine and internal standard, chlorpromazine, were extracted from human serum by liquid-liquid extraction with n-hexane containing 0.8% isopropanol and analyzed on a Capcell Pak CN column with the mobile phase of acetonitrile-0.2 M potassium dihydrogen phosphate (42:58, v/v, adjusted to pH 6.0 with 1 M NaOH). Detection wavelength of 251 nm and flow rate of 0.9 mL/min were fixed for the study. The assay robustness for the changes of mobile phase pH, organic solvent content, and flow rate was confirmed by $3^{3}$ factorial design using a fixed promethazine concentration (10 ng/mL) with respect to its peak area and retention time. In addition, the ruggedness of this method was investigated at three different laboratories using same quality control (QC) samples. This method showed linear response over the concentration range of 1-40 ng/mL with correlation coefficients greater than 0.999. The lower limit of quantification using 1 mL of serum was 1 ng/mL, which was sensitive enough for pharmacokinetic studies. The overall accuracy of the quality control samples ranged from 96.15 to 105.40% for promethazine with overall precision (% C.V.) being 6.70-11.22%. The relative mean recovery of promethazine for human serum was 63.54%. Stability (freeze-thaw and short-term) studies showed that promethazine was stable during storage, or during the assay procedure in human serum. However, the storage at $-80^{\circ}C$ for 4 weeks showed that promethazine was not stable. Extracted serum sample and stock solution were not allowed to stand at ambient temperature for 12 hr prior to injection. The peak area and retention time of promethazine were not significantly affected by the changes of mobile phase pH, organic solvent content, and flow rate under the conditions studied. This method showed good ruggedness (within 15% C.V.) and was successfully used for the analysis of promethazine in human serum samples for the pharmacokinetic studies of orally administered Himazin tablet (25 mg as promethazine hydrochloride) at three different laboratories, demonstrating the suitability of the method.

Validation of enzyme immunoassay for the quantitative measurement of human IgG antibodies specific for Haemophilus influenzae Type b capsular polysaccharide (Haemophilus influenzae type b 피막 다당질 특이 인간 IgG 항체의 정량적 측정을 위한 enzyme immunoassay의 타당성 연구)

  • Kim, Kyung Hyo;Lim, Soo Young
    • Clinical and Experimental Pediatrics
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    • v.50 no.2
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    • pp.143-150
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    • 2007
  • Purpose : This study was conducted to validate enzyme immunoassay (EIA) for the quantitative measurement of human IgG antibodies specific for Haemophilus influenzae type b (Hib) capsular polysaccharide. Method : We evaluated specificity, repeatability, intermediate precision, accuracy, lower limit of quantification (LLOQ), and stability to validate standardized EIA for the quantitative measurement of human anti-polyribosylribitol phosphate (PRP) IgG antibodies. Results : The results indicated that this EIA showed specificity to HbO-HA antigen and repeatability and intermediate precision were within acceptance criteria (repeatability: $CV{\leq}15%$, intermediate precision: $CV{\leq}20%$). The EIA-derived results from this laboratory were equivalent to those obtained by the standard radioactive antigen binding assay (RABA) for quantitation of anti-PRP antibodies in the 28 sera. Spiking recovery result was within acceptance criteria ($100{\pm}20%$). The precision and accuracy of samples in LLOQ were from -14.7 to -4.7% in nominal values, which were within acceptance criteria (precision: $CV{\leq}25%$, accuracy: ${\pm}25%$). Freeze-thaw stability and short term temperature stability were within ${\pm}20%$ of acceptance criteria. Conclusions : The EIA which is performed at the Center for Vaccine Evaluation and Study Ewha Medical Research Institute, is an appropriate serologic assay which can be used for quantitation of anti-PRP IgG antibodies in human sera.

A Study on the Material Characteristics and the Welding Properties of 600MPa Grade Steel (SM 570 TMC) (600MPa급(SM 570 TMC) 강재의 소재 및 용접특성에 관한 연구)

  • Kim, Jong Rak;Kim, Sang Seup;Lee, Chul Ho;Lee, Eun Taik;Beak, Ki Youl
    • Journal of Korean Society of Steel Construction
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    • v.20 no.6
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    • pp.773-781
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    • 2008
  • As buildings are becoming taller and longer-spanned, the requirements of high-strength and reliable steels are becoming increasingly stringent. Structural steels, however, acquire significantly different mechanical properties when their strength becomes higher. In this study, the mechanical properties, welding characteristics, and conformities of the 600MPa-grade high-strength steel were tested. The 600MPa-grade steel plates exhibited stable criterion strengthvalues and showed low carbon equivalents (${\mathcal{Ceq}}$) and composition (${\mathcal{Pcm}}$) as well as excellent welding hardness. In the tensile strength test, all the specimens were found to have strengths of over 600MPa. In the Sharphy impact test, the impact-absorbed energy of the V-notch specimens was shown to be 47J at the KS limit. Moreover, the maximum hardness of the specimens in the weld-heat-affected zone at a normal temperature was the same as that before welding. Their weld metal properties, however, were found not to be as good as those of high-strength steel. As such, the details of high-strength steel must be determined.

Study on VHCF Fatigue Behaviors and UNSM Effects of Hydrogen Attacked STS 316L (수소취화된 스테인리스강 316L의 VHCF(Very High Cycle Fatigue) 피로특성과 UNSM 효과에 관한 연구)

  • Nahm, Seung-Hoon;Baek, Un-Bong;Suh, Chang-Min;Pyun, Young-Sik
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.41 no.11
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    • pp.1011-1020
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    • 2017
  • This study was conducted to investigate the material properties of stainless steel 316L specimens of untreated and UNSM treated material, hydrogen attacked material(100 bar, $300^{\circ}C$ at 120 h) and UNSM treated hydrogen attacked material at room temperature. Results demonstrated that the hydrogen attacked materials showed a tendency toward a slightly decreased fatigue strength, while the hydrogen embrittlement effect was smaller than the S-N curve of conventional untreated material. As compared to untreated material, the fatigue limit of the UNSM treated material increased by 43.8%, while it was 57.1% higher in the UNSM treated hydrogen attacked material than in untreated hydrogen attacked material. The plastic deformation layer was ${\sim}152{\mu}m$ thick, as confirmed by maps showing the level of local plastic deformation affected by the UNSM treatment in three ways: an image quality map, inverse pole figure map, and kernel average misorientation map captured via electron back scatter diffraction. Owing to hydrogen embrittlement, about 90% of surface cracks were smaller than the average grain size of $35{\mu}m$.

Assessment of the Habitability for a Cabinet Fire in the Main Control Room of Nuclear Power Plant using Sensitivity Analysis (민감도 분석을 이용한 원전 주제어실의 케비닛 화재에 대한 거주성 평가)

  • Han, Ho-Sik;Lee, Jae-Ou;Hwang, Cheol-Hong;Kim, Joosung;Lee, Sangkyu
    • Fire Science and Engineering
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    • v.31 no.2
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    • pp.52-60
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    • 2017
  • Numerical simulations were performed to evaluate the habitability of an operator for a cabinet fire in the main control room of a nuclear power plant presented in NUREG-1934. To this end, a Fire Dynamics Simulator (FDS), as a representative fire model, was used. As the criteria for determining the habitability of operator, toxic products, such as CO, were also considered, as well as radiative heat flux, upper layer temperature, smoke layer height, and optical density of smoke. As a result, the probabilities of exceeding the criteria for habitability were evaluated through the sensitivity analysis of the major input parameters and the uncertainty analysis of fire model for various fire scenarios, based on V&V (Verification and Validation). Sensitivity analyses of the maximum heat release rate, CO and soot yields, showed that the habitable time and the limit criterion, which determined the habitability, could be changed. The present methodology will be a realistic alternative to enhancing the reliability for a habitability evaluation in the main control room using uncertain information of cabinet fires.

Occupational Factors Influencing the Forklift Operators' Exposure to Black Carbon (지게차 운전원의 블랙카본(black carbon, BC) 노출에 영향을 미치는 직업적 요인)

  • Lee, Hyemin;Lee, Seunghee;Ryu, Seung-Hun;Park, Jihoon;Park, Dong-Uk
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.27 no.4
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    • pp.313-323
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    • 2017
  • Objectives: This study aimed to assess exposure to black carbon(BC) among forklift operators and to identify environmental and occupational factors influencing their BC exposure. Methods: We studied a total of 23 forklift operators from six workplaces manufacturing paper boxes. A daily BC exposure assessment was conducted during working hours from January to April 2017. A micro-aethalometer was used to monitor daily BC exposure, and information on work activities was also obtained through a time-activity diary(TAD) and interviews. BC exposure records were classified into four categories influencing BC exposure level: working environment, workplace, forklift operation, and job characteristics. Analysis of variance(ANOVA) was used to compare average BC exposure levels among the four categories and the relationships between potential factors and BC exposure were analyzed using a multiple linear regression model. Results: The operators' daily exposure was $12.9{\mu}g/m^3$(N=9,148, $GM=7.5{\mu}g/m^3$) with a range: $0.001-811.4{\mu}g/m^3$. The operators were exposed to significantly higher levels when they operate a forklift in a room ${\leq}20,000m^3$($AM=12.3{\mu}g/m^3$), in indoor workplaces($AM=16.3{\mu}g/m^3$), when they operate a forklift manufactured before 2006 ($AM=13.2{\mu}g/m^3$), a forklift with a loading limit of four-tons($AM=27.1{\mu}g/m^3$), with a roll and bale type clamp($AM=17.1{\mu}g/m^3$), and with no particulate filter($AM=15.7{\mu}g/m^3$). Conclusions: Occupational factors including temperature, smoking, season, daytime, room volume($m^3$), location of operating, and manufacturing era and model of forklift influenced the BC exposure of forklift operators. The results of this study can be used to minimize the BC exposure of forklift operators.

Optimization of Indole-3-Acetic production by phosphate solubilization bacteria isolated from waste mushroom bed of Agaricus bisporus

  • Walpola, Buddhi Charana;Noh, Jae-Geun;Kim, Chan Kyem;Kyung, Ki-Cheon;Kong, Won-Sik;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.53-62
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    • 2013
  • A total of 35 phosphate solubilizing bacterial strains were isolated from waste mushroom bed of Agaricus bisporus in Buyeo-Gun, Chungnam and screened for the production of indole acetic acid (IAA). The best IAA producing strain was identified as Pantoea rodasii using 16S rRNA analysis. In addition to the IAA production, this strain could act as an efficient phosphate solubilizer (1100 ${\mu}g$ $ml^{-1}$ after 5 days of incubation) also. The selected strain was cultured under different conditions in order to assess the optimum conditions for maximum IAA production. The nutrient broth (NB) medium was recorded as the best medium, where the maximum IAA production (229 ${\mu}g$ $ml^{-1}$) was recorded at the start of stationary phase (12 hours after inoculation) of the bacteria growth. The performance of the strain was found to be maximum at the temperature of $30^{\circ}C$ followed by $25^{\circ}C$. IAA production was found to be increased with increasing tryptophan concentration (from 0.1 to 0.6%), however beyond this limit, a slight reduction in IAA production was observed. The strains' ability to produce IAA was further confirmed by extraction of crude IAA and subsequent TLC analysis. A specific spot from the extracted IAA preparation was found corresponding with the standard spot of IAA with same $R_f$ value. The results of HPLC analysis conducted in identifying and quantifying the IAA production more precisely, are in agreement with the results of the assessment done with colorimetric method. As revealed by the results of the pot experiment, the isolated strain could significantly enhance the growth (as measured by shoot and root growth) of mung bean plants compared to that of non-inoculated plants. Therefore it can be concluded that the present strain, Pantoea rodasii has great potential to be used as bio-inoculants.

Rapid Detection for Shiga Toxin Type 1 (Stxl) by Using Two-Step Ultra-Rapid Real-Time (URRT) PCR (초고속 이단계 PCR에 의한 Shiga 독소 타입 1의 신속 검출법)

  • Kim, Il-Wook;Kang, Min-Hee;Kwon, Soon-Hwan;Cho, Seung-Hak;Yoon, Byoung-Su
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.203-211
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    • 2008
  • Rapid detection-method for Shiga toxin type 1 that was produced from Shiga toxin-producing Escherichia coli (STEC) was developed by two-step ultra-rapid real-time (URRT) PCR. The specific primers were deduced from 80 bp stable region of stx type 1 (stxl) gene among various informations of STEC strains. URRT PCR is a microchip-based real-time PCR using 6 ${\mu}l$ of reaction volume with extremely short denaturation step and annealing/extension step (1 sec, 3 sec, respectively) in each cycle of PCR. Using the stx1-specific URRT PCR, 35 cycled PCR were finished in time of 6 min and 38 see, also measured 7 min and 28 see including melting temperature (Tm) analysis. The detection-limit of stxl-specific URRT-PCR was estimated until 3 colony forming units / PCR with products with stable Tm at $81.42{\pm}0.34^{\circ}C$. In the applications to various STEC strains and contaminated genomic DNAs, stx1-specific URRT-PCR were tested and shown that it would be expected an useful method for the rapid detection of stx1-coded STEC strains.

Applied Analysis for Metabolic Profiling of Trace-level Amino Acid in Biological Fluid (생체시료 중 미량 아미노산 대사 프로필을 위한 분석법 응용)

  • Nam, Hyung Wook;Park, Song-Ja;Pyo, Hee Soo;Paeng, Ki Jung
    • Analytical Science and Technology
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    • v.16 no.5
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    • pp.349-357
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    • 2003
  • The universality of low molecular weight metabolites (i.e. amino acids, steroid hormones) allows rapid and straightforward investigation of biochemistry of genetically un-characterized species. Thus in vivo metabolic profiling of amino acid in combination with multivariate data analysis (metabolomics) offers great potential in comparative biology. In this paper, amino acid profiles in biological fluid (media) were studied by using HPLC/FLD. HPLC procedure for amino acids require the formation of derivatives due to the low absorption of the free compounds. o-Phthalaldehyde (OPA) used in association with a thiol, such as 3-mercaptopropionic acid (3-MPA), is one of the most popular and sensitive reagents, which yield quickly fluorescent iso-indoles at room temperature. To improve unstability of OPA/3-MPA derivatization, we optimized injector programs for fixed injection times. Linear regressions for the standard curves were linear in the range 0.5 - 100.0 ppb, giving correlation coefficents above 0.99. The detection limit were 1.70 pmol(GLU) - 23.81 pmol(SER). It is practically useful when the amount of sample is very low on single cells.

Determination of Amitrole in Water Samples by Evaporation and GC/MS (증발법과 GC/MS를 이용한 수질 시료 중의 Amitrole의 분석법 연구)

  • Yoon, So Hee;Hong, Ji Eun;Pyo, Hee Soo;Park, Song Ja
    • Analytical Science and Technology
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    • v.16 no.6
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    • pp.483-487
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    • 2003
  • Amitrole is well-known as a non-selective herbicide and it is able to cause contamination of driking water as well as pollution of ground water and surface water. However, it is difficult to extract from water because it has a high solubility for water whereas a low solubility for general organic solvents. This method is described for the determination of amitrole in water samples by GC/MS. After evaporation of 10 mL water sample by a vacuum evaporator, amitrole was derivatized with isobutyl chloroformate (iso-BCF) on room temperature for 15~20 min. As a result, the sensitivity for GCfMS was improved as N-isobutoxycarbonyl amitrole derivative was formed. The linearity of the calibration curve showed good as 0.997. The recoveries were obtained more than 94.9% and relative standard deviations were less than 2.8% at $1.0{\mu}g/L$, $10.0{\mu}g/L$ and $100.0{\mu}g/L$. The limit of detection showed $0.1{\mu}g/L$ with a signal-to-noise ratio (S/N) of 3.