• Title/Summary/Keyword: Analytical Tool

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Bioluminescence immunoassay for neurotransmitter, serotonin using aequorin as a Label (Aequorin을 표지물질로 사용한 신경전달물질, 세로토닌에 대한 생물발광면역분석법)

  • Ryu, Ji-Eun;Choi, Hee-Seon;Park, Ho-Young;Rhee Paeng, In-Sook
    • Analytical Science and Technology
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    • v.23 no.1
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    • pp.60-67
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    • 2010
  • A sensitive competitive heterogeneous bioluminescence immunoassay for serotonin was developed using photoprotein, aequorin as a label for the first time with the optimal assay conditions; especially, serotoninavidin conjugate was prepared by Mannich reaction and the synthetic process of serotonin-avidin conjugate was optimized by controlling the initial molar ratios of serotonin, formaldehyde and avidin (1:12,000:25). The developed bioluminescence immunoassay for serotonin showed good sensitivity (LOD of 0.68 ng/mL) with wide area of dynamic range ($5.0{\times}10^{-10}\;M\sim5.0{\times}10^{-7}\;M$). (cf. the range for serotonin in human blood serum is $151{\pm}45\;ng$/mL). In addition, cross-reactivity studies demonstrated that 5-methoxytryptamine showed some cross-reactivity (28.0%), whereas 3-methylindole, melatonin and 5-hydroxylindole-3-acetic acid showed no crossreactivity, and good recoveries were obtained in serum. Thus, this developed method provides a good tool to monitor serotonin in serum.

Development of Dissolution Test Method for Acebrophylline Capsules and Bromhexine Hydrochloride Tablets in Korean Pharmaceutical Codex (고시수재 의약품 중 아세브로필린 캡슐 및 브롬헥신염산염 정의 용출시험법 개발)

  • Lee, Tae-Woong;Jeong, Rae-Seok;Jeong, Seung-A;Kim, Jeong-Hyun;Shim, Young-Hun;Kim, In-Kyu;Park, Chang-Won
    • YAKHAK HOEJI
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    • v.57 no.3
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    • pp.226-233
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    • 2013
  • Although the dissolution test can serve as an effective tool for quality control and predictor of in vivo performance, there are a number of drugs with no established dissolution specification in Korean Pharmaceutical Codex (KPC). So, with each reference and test drugs, the dissolution test method and an analytical procedure by HPLC were developed and validated to establish dissolution specification for acebrophylline capsules and bromhexine hydrochloride tablets. The dissolution condition was determined based on the "Guidelines on Specifications of Dissolution tests for Oral dosage forms" of Ministry of Food and Drug Safety (MFDS). The analytical method of HPLC was validated in specificity, linearity, precision and accuracy. Final dissolution test was performed with commercially available samples of 3 lots to establish specification. In addition, no difference was observed by the inter-laboratory evaluation. Dissolution specifications and conditions will be used for revising the monograph of acebrophylline capsules and bromhexine hydrochloride tablets in next supplement of KPC.

Functional Integration of Serial Dilution and Capillary Electrophoresis on a PDMS Microchip

  • Chang, Jun-Keun;Heo, Yun-Seok;Hyunwoo Bang;Keunchang Cho;Seok Chung;Chanil Chung;Han, Dong-Chul
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.4
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    • pp.233-239
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    • 2003
  • For the quantitative analysis of an unknown sample a calibration curve should be obtained, as analytical instruments give relative, rather than absolute measurements. Therefore, researchers should make standard samples with various known concentrations, measure each standard and the unknown sample, and then determine the concentration of the unknown by comparing the measured value to those of the standards. These procedures are tedious and time-consuming. Therefore, we developed a polymer based microfluidic device from polydimethylsiloxane, which integrates serial dilution and capillary electrophoresis functions in a single device. The integrated microchip can provide a one-step analytical tool, and thus replace the complex experimental procedures. Two plastic syringes, one containing a buffer solution and the other a standard solution, were connected to two inlet holes on a microchip, and pushed by a hydrodynamic force. The standard sample is serially diluted to various concentrations through the microfluidic networks. The diluted samples are sequentially introduced through microchannels by electro-osmotic force, and their laser-induced fluorescence signals measured by capillary electrophoresis. We demonstrate the integrated microchip performance by measuring the fluorescence signals of fluorescein at various concentrations. The calibration curve obtained from the electropherograms showed the expected linearity.

One-Step Simultaneous Immunochromatographic Strip Test for Multianalysis of Ochratoxin A and Zearalenone

  • Shim, Won-Bo;Dzantiev, Boris B.;Eremin, Sergei A.;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.19 no.1
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    • pp.83-92
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    • 2009
  • Individual immunochromatographic assays (ICG) for ochratoxin A (OTA) and zearalenone (ZEA) were optimized and used in the development of a one-step simultaneous immunochromatographic assay (OS-ICG) for the rapid multianalysis of two mycotoxins in corn samples. The nitrocellulose membrane of the OS-ICG was treated with OTA-bovine serum albumin (BSA), ZEA-ovalbumin (OVA), and anti-mouse IgG in the OTA test, ZEA test, and control zones, respectively. Monoclonal antibody-gold conjugates (OTA3 MAb-gold and ZEA2C5 MAb-gold) were sprayed onto the conjugate pad. The visual detection limits were 2.5 and 5 ng/ml for OTA and ZEA, respectively, and the results were obtained within 15 min after starting the analysis. An efficient, simple, and rapid extraction method using 30% MeOH/PBS was established and validated by analyzing the corn samples spiked with OTA/ZEA mixtures (0/0, 5/10, 10/20, and $20/30\;{\mu}g/kg$). The cut-off values of the OS-ICG for the spiked corn were 5 and $10\;{\mu}g/kg$ for OTA and ZEA, respectively. Natural corn samples were analyzed by OS-ICG, direct competitive enzyme-linked immunosorbent assay (DC-ELISA), and HPLC. Results of the OS-ICG were in good agreement with those obtained by DC-ELISA and HPLC. The developed OS-ICG offers a rapid, easy-to-use, and portable analytical system and can be used as a convenient qualitative tool for the on-site simultaneous determination of OTA and ZEA in cereals, food, and agricultural products in one analytical cycle.

A Numerical Analysis of Land Use-Transportation Model as a Form of Analytical Tool (수치해석적 토지이용-교통모형의 이론연구 도구화: 교통수요의 내생화를 중심으로)

  • Yu, Sang-Gyun;Rhee, Hyok-Joo
    • Journal of Korean Society of Transportation
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    • v.31 no.2
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    • pp.33-44
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    • 2013
  • The land use-transportation models typically have complicated model structure that is good for empirical execution but bad for theoretical probe. This complexity makes it very difficult to derive the first-order conditions for system optimization in tractable forms. Yu and Rhee (2011) and Rhee (2012) show how to simplify the derivative of the model's objective function with respect to policy variables in the computable general equilibrium model of land use and transportation. However, the travel demand in their model was fixed. This drawback fundamentally limits the applicability of their methodology in the planning field. We relax this restriction. Once this is done, we can employ the methodology developed in analyzing the impacts of various types of policy instruments in the models where land market is treated endogenously and transportation network is embedded.

Observation of the silicon acrylate effect on the photo-polymerization reaction using micro raman spectroscopic technique (마이크로 라만을 사용한 실리콘 아크릴레이트가 광중합 반응에 미치는 영향 관찰)

  • Oh, HyangRim;Hong, Jin-Who;Yu, Jeong-A
    • Analytical Science and Technology
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    • v.17 no.3
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    • pp.225-229
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    • 2004
  • The effect of the silicon acrylate as a reactive additive on the UV-curing photopolymerization reaction was studied by micro raman technique. For the study, acrylate systems and Darocur 1173 were used as oligomer and monomers, and a photo initiator, respectively. The content of silicon acrylate was within the range of 0-3 wt%. The extent of photo-polymerization reaction as a function of depth from the air interface was obtained from the conversion ratio of acrylate double bond calculated from the intensities of measured bands at $1410cm^{-1}$ and at $1635cm^{-1}$. Micro raman spectroscopic technique can be an useful tool for the investigation of the factors, which can affect the reaction progress, such as oxygen inhibition, composition of the formulations, depth, etc.

Toward High-Resolution Cryo-Electron Microscopy: Technical Review on Microcrystal-Electron Diffraction

  • Lee, Sangmin;Chung, Jeong Min;Jung, Hyun Suk
    • Applied Microscopy
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    • v.47 no.4
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    • pp.223-225
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    • 2017
  • Cryo-electron microscopy (cryo-EM) is arguably the most powerful tool used in structural biology. It is an important analytical technique that is used for gaining insight into the functional and molecular mechanisms of biomolecules involved in several physiological processes. Cryo-EM can be separated into the following three groups according to the analytical purposes and the features of the biological samples: cryo-electron tomography (cryo-ET), cryo-single-particle reconstruction, and cryo-electron crystallography. Cryo-tomography is a unique EM technique that is used to study intact biomolecular complexes within their original environments; it can provide mechanistic insights that are challenging for other EM-methods. However, the resolution of reconstructed three-dimensional (3D) models generated by cryo-ET is relatively low, while single-particle reconstruction can reproduce biomolecular structures having near-atomic resolution without the need for crystallization unless the samples are large (>200 kDa) and highly symmetrical. Cryo-electron crystallography is subdivided into the following two categories according to the types of samples: one category that deals with two-dimensional (2D) crystalline arrays and the other category that uses 3D crystals. These two categories of electron-crystallographic techniques use different diffraction data obtained from still diffraction and continuous-rotation diffraction. In this paper, we review crystal-based cryo-EM techniques and focus on the recently developed 3D electron-crystallographic technique called microcrystal-electron diffraction.

Animal species identification by co-amplification of hypervariable region 1 (HV1) and cytochrome b in mitochondrial DNA

  • Lim, Si Keun;Park, Ki Won
    • Analytical Science and Technology
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    • v.18 no.3
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    • pp.257-262
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    • 2005
  • Mitochondrial DNA (mt DNA) sequence analysis has been a useful tool for species identification of animals and human individuals. Two hypervariable regions (HV1 and HV2) in control region of mitochondrial genome were analyzed for human individual identification. In case of animal species identification, several genes on mt DNA such as cytochrome b (cytb), RNAs, cytochrome oxidases (CO) were used. In this study, co-amplification of HV1 and cytb was carried out in order to check the contamination of animal DNA and to verify the human DNA. The primer sets used in PCR were H15997/L16236 for HV1 and H14724/L15149 for cytb. PCR products for HV1 and cytb were 239 bp and 425 bp, respectively. The appearance of two bands on agarose gel implied the DNA came from human, however the single band of cytb gene represented the non-human animal DNA.

Comparison of Cell Lysis Techniques via Q-TOF LC/MS

  • Kaplan, Ozan;Oncul, Selin;Ercan, Ayse;Celebier, Mustafa
    • Mass Spectrometry Letters
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    • v.11 no.2
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    • pp.36-40
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    • 2020
  • Untargeted metabolomics is a useful tool for drug development focusing on novel chemotherapeutic and chemopreventative agents against cancer cells. In recent years, quadrupole time of flight liquid chromatography-mass spectrometry (Q-TOF LC/MS)-based untargeted metabolomic approaches have gained importance to evaluate the effect of these agents at the molecular level. The researchers working on cell culture studies still do not apply standardized methodologies on sample preparation for untargeted metabolomics approaches. In this study, the rough and wet lysis techniques performed on MCF-7 breast cancer cells were compared with each other via the Q-TOF LC/MS-based metabolomic approach. The C18 and hydrophilic interaction liquid chromatography (HILIC) columns were used for the separation of the metabolites in MCF-7 cell lysates. 505 peaks were detected through the HILIC column and 551 peaks were found through the C18 column for the wet lysis technique. This situation supported by the base peak chromatograms showed that the wet lysis technique allowed us to extract higher number of non-polar metabolites. Almost equal number of metabolites was found for the C18 and HILIC columns (697 peaks for the HILIC column and 695 peaks for the C18 column) when the rough lysis technique was used. However, the intensities of polar metabolites were higher for the rough lysis technique on base peak chromatograms for both the HILIC and C18 columns. Although cell lysis technique, which is the first step in the sample preparation for cell culture studies, does not cause dramatic differences in the number of the detected metabolite peaks, it affects the polar and non-polar metabolite ratio significantly. Therefore, it must be considered carefully especially for in vitro drug development studies.

Hands-on Tools to Prevent Human Errors in Highway Construction (고속도로 건설현장의 인적오류 예방을 위한 실무자용 도구 개발)

  • Kim, Jung-Yong;Yoon, Sang-Young;Cho, Young-Jin
    • Journal of the Ergonomics Society of Korea
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    • v.30 no.1
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    • pp.19-28
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    • 2011
  • Objective: The aim of this study is to reclassify human errors and to develop hands-on tools to apply the new classification for preventing human error accidents in highway construction site. Background: The main cause of accidents in highway construction was reported as the carelessness of workers. However, such diagnosis could not help us operationally prevent accidents in real workplace. Method: The accidents in highway construction were reanalyzed and the causes of human error were reclassified in order to educate and improve the awareness of human error in highway construction. Field survey and interview with safety managers and workers were conducted to find the causal relationship between the actual accidents and the human errors. Results: The most frequently observed human errors in highway construction were classified into six categories such as mis-perception, distraction, memory fail, slip, cognition error and mis-judgment. In order to provide hands-on tools to increase the awareness of human error in construction field, the human error checklist and card sorting diary were developed. Especially, the card sorting diary was designed to increase the ability in human error inspection of safety manager at construction site. Moreover, posters were developed based on actual accident cases. Conclusion: We suggested that the improved awareness and analytical report on checklist, card sorting diary and posters for construction field could collectively prevent the accident. Application: The classification of human error, hands-on tools and posters can be directly applicable on highway construction site. This analytical and collective approach preventing human error-related accident could be extended to other construction workplaces.