• Title/Summary/Keyword: Purification method

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Effect of Antibody Immobilization Method to Magnetic Micro Beads on its Immunobinding Characteristics (자성 미세입자에의 항체 고정화 방법이 면역결합반응에 미치는 영향)

  • Choi, Hyo Jin;Hwang, Sang Youn;Jang, Dae Ho;Cho, Hyung Min;Kang, Jung Hye;Seong, Gi Hun;Choo, Jae Bum;Lee, Eun Kyu
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.65-72
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    • 2006
  • Recent technical advances in the biorecognition engineering and the microparticle fabrication may enable us to develop the single step purification using magnetic particle, because of its simplicity, efficacy, ease of automation, and process economics. In this study, we used commercial magnetic particles from Seradyn, Inc. (Indianapolis, USA). It was ca. 2.8 micron in diameter, consisted of polystyrene core and magnetite coating, and its surface had carboxyl groups. The model, capture protein was IgG and anti-IgG was used as the ligand molecule. We studied the different surfaces ('nude', ester-activated, and anti-IgG coated) for their biorecognition of IgG. At a high pH condition, we could reduce non-specific binding. Also anti-IgG immobilized magnetic particle could capture IgG more selectively. We attempted 'oriented immobilization' of anti-IgG, in which the polysaccharides moiety near the C-terminus was selectively oxidized and linked to the hydrazine-coated MP, to improve the efficacy of biorecognitive binding. Using this method, the IgG capturing ability was improved by ca. 2 fold. From the binary mixture of the IgG-insulin, IgG could be more selectively captured. In summary, the oriented immobilization of oxidized anti-IgG proved to be as effective as the streptavidin-biotin system and yet simpler and cost-effective. This immobilization method can find its applications in protein biochips and biotargeting.

Study of Utilization of Natural Zeolites as Functional Materials for Water Purification (I): Cation Exchange Property of Domestic Zeolites (천연 제올라이트의 수환경 개선용 기능성 소재로의 활용에 관한 연구 (I): 국내산 제올라이트의 양이온 교환 특성)

    • Journal of the Mineralogical Society of Korea
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    • v.16 no.2
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    • pp.135-149
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    • 2003
  • Domestic zeolite ores are mostly composed of Ca-type clinoptilolite, accompanying a little amounts of mordenite. However, other types of zeolite ores rich in ferrierite, heulandite, or mordenite are less commonly found. Based on the quantitative XRD analysis, zeolite contents are determined to be nearly 50∼90 wt%. Impurities (mostly > 10 wt%) in the zeolite ores chiefly consist of quartz, feldspar, smectite, and opal-CT. The determined CEC values ($CEC_{AA}$ ) of powdery samples (grain size: < 125 $\mu\textrm{m}$) of zeolite ores by the Ammonium Acetate method are mostly higher than 100 meq/100 g. Some zeolites from the Guryongpo area, corresponding to the clinoptilolite ore, are measured to be dominantly high in CEC values ranging 170∼190 meq/100 g. Cation exchange property of the zeolite ores varies greatly depending on the types or zeolite species present in the ores. Despite of the lower grade in zeolite content, the $CEC_{AA}$ of ferrierite ore is comparatively high. Compared to this, the $CEC_{AA }$ of heulandite ore is very low, though the zeolite ore exhibits the highest grade ranging up to about 90 wt%. In addition, the CEC values calculated theoretically from the framework composition of clinoptilolite-heulandite series are not consistent with those determined by the cation exchage experiment. The measured $CEC_{AA}$ of clinoptilolite ores are generally higher than those of heulandite ores. This may be due to the higher Ca abundance in exchangeable cation composition and the presence of probable stacking faults in heulandite. The variation of $CEC_{CEC}$ is roughly proportional, though not strictly compatible, to the zeolite contents in clinoptilolite ores. It seems to be caused by the fact that the $CEC_{AA}$ of clinoptilolite locally varies depending on crystal-chemical diversity, i. e., the variation in framework composition (Si/Al) and exchangeable cation composition (especially, the contents of Ca and K). In addition, the determined CEC values ($CEC_{MB}$ ) of zeolite ores by the Methylene Blue method are much higher than those calculated from smectite contents. It suggests a probable reaction of Methylene Blue ion ($C_{16}$ $H_{18}$ $N_3$S+) with larger-pore zeolites than clinoptlolite-heulandite series, i.e., ferrierite and mordenite as well as with smectite. This can be supported by the fact that the ferrierite ore accompanying little amount of smectite has the highest value in CE $C_{MB}$ .

Synthesis of Visible-working Pt-C-TiO2 Photocatalyst for the Degradation of Dye Wastewater (염료폐수 분해를 위한 가시광 감응형 Pt-C-TiO2 광촉매의 합성)

  • Hahn, Mi Sun;Yun, Chang Yeon;Yi, Jongheop
    • Clean Technology
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    • v.11 no.3
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    • pp.123-128
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    • 2005
  • Among various metal oxides semiconductors, $TiO_2$ is the most studied semiconductor for environmental clean-up applications due to its unique ability in photocatalyzing various organic contaminants, its chemical inertness, and nontoxicity. $TiO_2$, however, has a few drawbacks to be solved such as reactivity mainly working under ultraviolet irradiation (${\lambda}$ < 387 nm) and electron - hole recombination on $TiO_2$. In this study, to extend the absorption range of $TiO_2$ into the visible range and enhance electron - hole separation, we synthesized platinum (Pt) deposited $C-TiO_2$. The presence of Pt as an electron sink has been known to snhance the separation of photogenerated electron-hole pairs and induce the thermal decomposition. The characterization of as-synthesized $Pt-C-TiO_2$ was performed by Transmission Electron Microscopic (TEM), the Brunuer-Emmett-Teller (BET) method, X-ray Diffractometer (XRD), UV-vis spectrometer (UV-DRS), and X-ray Photoelectron Spectroscopy (XPS). In order to estimate the photocatalytic activity of the synthesized materials, the photoelectron Spectroscopy (XPS). In order to estimate the photocatalytic activity of the synthesized materials, the photodegradation experiment of an azo dye (Acid Red 44; $C_{10}H_7N=NC_{10}H_3(SO_3Na)_2OH$)was carried out by using an Xe arc lamp (300 W, Oriel). A 420 nm cut-off filter was used for visible light irradiation. From the results, Pt-deposited $C-TiO_2$ showed a far superior phothdegradation activity to Degussa P25, the commercial product under the irradiation of visible light and enhanced photocatalytic activity of visible-working $C-TiO_2$. This is a useful result into the application for the purification system of dye wastewater using visible energy of sun light.

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Determination of Carazolol and Azaperone in Livestock and Fishery Products Using Liquid Chromatography-tandem Mass Spectrometry (축수산물에서 LC-MS/MS를 이용한 카라졸롤 및 아자페론 분석)

  • Choi, Soo Yeon;Kang, Hui-Seung;Kim, Joohye;Cheon, So-Young;Jeong, Jiyoon;Cho, Byung-Hoon;Lee, Kang-Bong
    • Journal of Food Hygiene and Safety
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    • v.33 no.3
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    • pp.176-184
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    • 2018
  • The aim of the present work was to develop simultaneous methods of quantification of carazolol, azaperone, and azaperol residues in livestock and fishery products using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Samples were extracted from beef, pork, chicken, egg, milk and shrimp using acetonitrile (ACN); while flat fish and eel were extracted using 80% ACN. For purification, ACN saturated n-hexane was used to remove fat composition. The standard calibration curves showed good linearity as correlation coefficients; $r^2$ was > 0.99. Average recoveries expressed were within the range of 67.9-105% for samples fortified at three different levels ($0.5{\times}MRL$, $1{\times}MRL$ and $2{\times}MRL$). The correlation coefficient expressed as precision was within the range of 0.55-7.93%. The limit of quantification (LOQ) was 0.0002-0.002 mg/kg. The proposed analytical method showed high accuracy and acceptable sensitivity based on Codex guideline requirements (CAC/GL71-2009). This method can be used to analyze the residue of carazolol, azaperone, and azaperol in livestock and fishery products.

Mechanisms of Lipopolysaccharide-induced Lipopolysaccharide Tolerance in the Expression of TNF-$\alpha$ and IL-8 in Peripheral Blood Monocytes (말초 혈액 단핵구의 TNF-$\alpha$와 IL-8 발현에서 내독소에 대한 내성 기전에 관한 연구)

  • Park, Gye-Young;Kim, Jae-Yeol;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.601-610
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    • 1997
  • Background : Monocytes/macrophages play a central role in determining the host response during Gram-negative infection through secretion of a variety of mediators after stimulation of LPS. Even though cytokine production has been shown to play an important role in host defense during sepsis, cytokine release may also lead to tissue injury. Thus, regulation of macrophage response to LPS is critical for host survival during Gram-negative sepsis. In animals exposed to nonlethal doses of endotoxin, a characteristic hyporesponsiveness to subsequent administration of endotoxin has been observed. This phenomenon was known as 'LPS tolerance'. However, little information is available regarding the underlying mechanism of LPS tolerance. Method : Peripheral blood monocyte(PBMC) was isolated from peripheral blood of normal volunteers by adhesion purification method. To evaluate the conditions to obtain LPS tolerance, preculture was carried out with LPS at 10ng/ml for 24 hours. For stimulation, culture plates were washed two times and were stimulated with LPS at $1{\mu}g/ml$ for 4, 6 and 26 hours. To assess the underlying mechanisms of LPS tolerance, autologous serum, PMA, anti-CD14 Ab, Indomethacin or $PGF_2$ were added to preculture solution respectively. Cytokine concentrations in culture supernatants were measured using ELISA for TNF-$\alpha$ and IL-8 and mRNA of TNF-$\alpha$ and IL-8 were determined by Northern blot analysis. Results : The exposure of PBMC to low dose of LPS suppressed the cytokine production and mRNA expression of TNF-$\alpha$, but not IL-8. Anti-CD14 Ab partially recovered production of TNF-$\alpha$ which was suppressed by preculture with low dose LPS. The preculture with PMA induces LPS tolerance, as preculture with low dose LPS. Conclusion : LPS tolerance to TNF-$\alpha$ is regulated pretranslationally and is influenced by protein kinase C pathway and CD14.

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A Novel Glycine-Rich Region in Sox4 is a Target for the Proteolytic Cleavage in E. coli (전사활성 인자인 Sox4의 단백질 분해효소에 의한 표적 부위에 관한 연구)

  • 허은혜;최주연;장경희;김인경;임향숙
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.153-161
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    • 2002
  • Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

Optimum Pre-treatment Method in Constructed Wetlands by Natural Purification Method for Treating Livestock Wastewater (자연정화공법에 의한 인공습지에서 효과적인 축산폐수처리를 위한 최적 전처리방법 구명)

  • Park, Jong-Hwan;Seo, Dong-Cheol;Kim, Ah-Reum;Kim, Sung-Hun;Lee, Seong-Tea;Jeong, Tae-Uk;Choi, Jeong-Ho;Lee, Sang-Won;Cho, Ju-Sik;Kim, Hyun-Ook;Heo, Jong-Soo
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.3
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    • pp.425-433
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    • 2011
  • In order to obtain optimum pre-treatment methods and improve T-N and T-P removal efficiencies, removal rates of pollutants in small-scale livestock wastewater treatment apparatus with water plant filtration bed or activated sludge tank were investigated. Based on the results from the optimum pre-treatment in small-scale livestock wastewater treatment apparatus, removal efficiencies of pollutants in livestock wastewater treatment plant with water plant filtration and activated sludge beds. The removal rates of COD, SS, T-N, and T-P in effluent were 83, 89, 63 and 87% in small-scale livestock wastewater treatment apparatus with water plant filtration bed, respectively. The removal rates of COD, SS, T-N, and T-P in effluent were 96, 95, 86 and 92% in small-scale livestock wastewater treatment apparatus with activated sludge tank, respectively. For increasing the COD, SS, T-N, and T-P removals in small-scale livestock wastewater treatment apparatus, the water plant filtration and activated sludge beds are recommended. In livestock wastewater treatment plant with water plant filtration ($1^{st}$ treatment) and activated sludge ($2^{nd}$ treatment) beds, the concentrations of COD, SS, T-N, and T-P in effluent were 39, 15, 42 and $1mg\;L^{-1}$, respectively. It was shown that the concentrations of COD, SS, T-N, and T-P met acceptable effluent quality standard for livestock wastewater. Based on the above results, the removal rates of COD, SS, T-N, and T-P in effluent were over 99.8, 99.9, 99.2, and 99.9% in livestock wastewater treatment plant, respectively.

The control of TiO2 nanofiber diameters using fabrication variables in electrospinning method (전기 방사 공정의 제조 변수를 이용한 TiO2 나노섬유의 직경 제어)

  • Yoon, Han-Sol;Kim, Bo-Sung;Kim, Wan-Tae;Na, Kyeong-Han;Lee, Jung-Woo;Yang, Wan-Hee;Park, Dong-Cheol;Choi, Won-Youl
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.31 no.1
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    • pp.8-15
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    • 2021
  • TiO2 has been used in various fields such as solar cells, dental implants, and photocatalysis, because it has high physical and chemical stability and is harmless to the body. TiO2 nanofibers which have a large specific surface area also show a good reactivity in bio-friendly products and excellent photocatalysis in air and water purification. To fabricate TiO2 nanofibers, an electrospinning method was used. To observe the diameter of TiO2 nanofibers with fabrication variables, the fabrication variables was divided into precursor composition variables and process variables and microstructure was analyzed. The concentrations of PVP (Polyvinylpyrrolidone) and TTIP (Titanium(IV) isopropoxide) were selected as precursor composition variables, and inflow velocity and voltage were also selected as process variables. Microstructure and crystal structure of TiO2 nanofibers were analyzed using FE-SEM (Field emission scanning electron microscope) and XRD (X-ray diffraction), respectively. As-spun TiO2 nanofibers with an average diameter of about 0.27 ㎛ to 1.31 ㎛ were transformed to anatase TiO2 nanofibers with an average diameter of about 0.22 ㎛ to 0.78 ㎛ after heat treatment of 3 hours at 450℃. Anatase TiO2 nanofibers with an average diameter of 0.22 ㎛ can be expected to improve the photocatalytic properties by increasing the specific surface area. To change the average diameter of TiO2 nanofibers, the control of precursor composition variables such as concentrations of PVP and TTIP is more efficient than the control of electrospinning process variables such as inflow velocity and voltage.

Development and Validation of an Analytical Method for Fungicide Fluoxastrobin Determination in Agricultural Products (농산물 중 살균제 Fluoxastrobin의 시험법 개발 및 유효성 검증)

  • So Eun, Lee;Su Jung, Lee;Sun Young, Gu;Chae Young, Park;Hye-Sun, Shin;Sung Eun, Kang;Jung Mi, Lee;Yun Mi, Chung;Gui Hyun, Jang;Guiim, Moon
    • Journal of Food Hygiene and Safety
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    • v.37 no.6
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    • pp.373-384
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    • 2022
  • Fluoxastrobin a fungicide developed from Strobilurus species mushroom extracts, can be used as an effective pesticide to control fungal diseases. In this study, we optimized the extraction and purification of fluoxastrobin according to its physical and chemical properties using the QuEChERS method and developed an LC-MS/MS-based analysis method. For extraction, we used acetonitrile as the extraction solvent, along with MgSO4 and PSA. The limit of quantitation of fluoxastrobin was 0.01 mg/kg. We used 0.01, 0.1, and 0.5 mg/kg of five representative agricultural products and treated them with fluoxastrobin. The coefficients of determination (R2) of fluoxastrobin and fluoxastrobin Z isomer were > 0.998. The average recovery rates of fluoxastrobin (n=5) and fluoxastrobin Z isomer were 75.5-100.3% and 75.0-103.9%, respectively. The relative standard deviations (RSDs) were < 5.5% and < 4.3% for fluoxastrobin and fluoxastrobin Z isomer, respectively. We also performed an interlaboratory validation at Gwangju Regional Food and Drug Administration and compared the recovery rates and RSDs obtained for fluoxastrobin and fluoxastrobin Z isomer at the external lab with our results to validate our analysis method. In the external lab, the average recovery rates and RSDs of fluoxastrobin and fluoxastrobin Z isomer at each concentration were 79.5-100.5% and 78.8-104.7% and < 18.1% and < 10.2%, respectively. In all treatment groups, the concentrations were less than those described by the 'Codex Alimentarius Commission' and the 'Standard procedure for preparing test methods for food, etc.'. Therefore, fluoxastrobin is safe for use as a pesticide.

A Study on Environmental Standards of School Building (교사환경기준에 관한 연구)

  • Hong, Seok-Pyo;Park, Young-Soo
    • The Journal of Korean Society for School & Community Health Education
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    • v.1 no.1
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    • pp.11-43
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    • 2000
  • The purpose of this study was, through analyzing the previous researches, to grasp the present status of environment of school building(ESB), research the sundry records of each element and, through comparative analysis of the standard of ESB in Korea, the United States, and Japan, select the normative standard of ESB, to clarify the point at issue presented in Regulation of Construction & facility Management for Elementary and and Secondary School in Korea, and to suggest an alternative preliminary standard of ESB. To carry out a research for this purpose, these were required: 1. to investigate the existing present status of ESB, 2. to make a comparative analysis of the standard of ESB in each country, 3. to suggest the normative standard of preliminary standard of ESB, 4. to analyze the controversial points of the standard of ESB in Korea, 5. to suggest an alternative preliminary standard of ESB. The conclusions were as follows: 1. Putting, through analyzing the previous researches, the existing present status of ESB together, it seemed that lighting environment, indoor air environment and noise environment were all in poor conditions. 2. In the result of a comparative analysis of the standard of ESB in Korea, Japan and the United States, in Korea the factors of each lighting and indoor air environment were not presented properly, in Japan, in lighting environment aspect, the standard on natural lighting and the factors on brightness were not presented., and in the USA the essential factors of each environment were throughly presented. In the comparison of the standards on each factor, Korea showed that the standard level presented was less properly prescribed than those of the USA and Japan but it also showed that the standard levels prescribed in the USA and in Japan were mostly similar to the standard levels in records investigated. 3. With the result of the normative standard selection on School Builiding environment factor of prescribed in this study, the controversial points of the standard of ESB in Korea were analyzed and the result was utilized to suggest new preliminary standard of ESB. 4. As the result of the analysis of the controversial points of the standard of ESB in Korea, it was found that the standard of ESB in Korea should be established on a basis of School Health Act and be concretely presented in School Health Regulation and School Health Rule. The factors of each environment was improperly presented in the existing standard of ESB in Korea. Moreover the standard of them was inferior to that of the records investigated and those of in the USA and in Japan and it also showed that the standard of it in Korea was improper to maintain Comfortable Learning Environment. 5. A suggested preliminary standard of ESB acquired through above study as follows: 1) In this study a new kind of preliminary standard of ESB is divided into lighting environment, indoor air environment, noise environment, odor environment and for above classification, reasonable factor and standard should be established and the controling way on each standard and countermeasures against it should be considered. 2) In lighting environment, the factors of natural lighting are divided into daylight rate, brightness, glare. In the standard on each factor, daylight rate should secure 5% of a mean daylight rate and 2% of a minimum daylight rate, brightness ratio of maximum illumination to minimum illumination should be under 10:1, and in glare there should not be an occurrence factor from a reflector outside of the classroom. And the factors of unnatural lighting are illumination, brightness, and glare. In the standard on each factor, illumination should be 750 lux or more, brightness ratio should be under 3 to 1, and glare should not occur. And Optimal reflection rate(%) of Colors and Facilities of Classroom which influences lighting environment should be considered. 3) In indoor air environment factors, thermal factors are divided into (1) room temperature, (2) relative humidity, (3) room air movement, (4) radiation heat, and harmful gases (5) CO, (6) $CO_2$ that are proceeded from using the heating fuel such as oval briquettes, firewood, charcoal being used in most of the classroom, and finally (7) dust. In the standard on each factor, the next are necessary; room temperature: $16^{\circ}C{\sim}26^{\circ}C$(summer : $E.T18.9{\sim}23.8^{\circ}C$, winter: $E.T16.7{\sim}21.7^{\circ}C$), relative humidity: $30{\sim}80%$, room air movement: under 0.5m/sec, radiation heat: under $5^{\circ}C$ gap between dry-bulb temperature and wet-bulb temperature, below 1000 ppm of ca and below 10ppm of $CO_2$, dust: below 0.10 $mg/m^3$ of Volume of dust in indoor air, and ventilation standard($CO_2$) for purification of indoor air : once/6 min.(about 7 times/40 min.) in an airtight classroom. 4) In the standard on noise environment, noise level should be under 40 dB(A) and the noise measuring way and the countermeasures against it should be considered. 5) In the standard on odor environment, odor level under Physical Method should be under 2 degrees, and the inspecting way and the countermeasures against it should be considered.

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