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Autometallography for Zinc Detection in the Central Nervous System (중추신경계통내 분포하는 Zinc의 조직화학적 동정)

  • Jo, Seung-Mook;Gorm, Danscher;Kim, Sung-Jun;Park, Seung-Kook;Kang, Tae-Cheon;Won, Moo-Ho
    • Applied Microscopy
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    • v.30 no.4
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    • pp.347-355
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    • 2000
  • Zinc is one of the most abundant oligoelements in the living cell. It appears tightly bound to some metalloproteins and nucleic acids, loosely bound to some metallothioneins or even as free ion. Small amounts of zinc ions (in the nanomolar range) regulate a plentitude of enzymatic proteins, receptors and transcription factors, thus rolls need accurate homeostasis of zinc ions. Zinc is an essential catalytic or structural element of many proteins, and a signaling messenger that is released by neural activity at many central excitatory synapses. Growing evidences suggest that zinc may also be a key mediator and modulator of the neuronal death associated with transient global ischemia and sustained seizures, as well as perhaps other neurological disease stoles. Some neurons have developed mechanisms to accumulate zinc in specific membrane compartment ('vesicular zinc') which can be evidenced using histochemical techniques. Substances giving a bright colour or emitting fluorescence when in contact with divalent metal ions are currently used to detect them inside cells; their use leads to the so called 'direct' methods. The fixation and precipitation of metal ions as insoluble salt precipitates, their maintenance along the histological process and, finally, their demonstration after autometallographic development are essential steps for other methods, the so called 'indirect methods'. This study is a short report on the autometallograhical approaches for zinc detection in the central nervous system (CNS) by means of a modified selenium method.

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Effects of Cyclobuxine E on Two Distinct Types of Potassium-Activated Calcium Channels in an Intestinal Smooth Muscle (흰쥐 장관에 있어 칼륨에 의해 활성화되는 칼슘 채널에 대한 Cyclobuxine E의 영향)

  • Lee, Jong-Hwoa;Kwon, Jun-Tack;Cho, Byung-Heon;Choi, Kyu-Hong;Kim, Yu-Jae;Kim, Jong-Bae;Kim, Chun-Sook;Cha, Young-Deog;Kim, Young-Suk
    • The Korean Journal of Pharmacology
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    • v.25 no.1
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    • pp.53-58
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    • 1989
  • For several years, we investigated the pharmacological action of several substances isolated from Buxus microphylla var koreana Nakai, which had been used as folk remedies of malaria and venereal disease. Cyclobuxine $D(C_{25}H_{42}ON_2)$, a steroidal alkaloid, exerted an antiinflammatory action, hypotensive and bradycardic effects in rats. In the present study, we isolated alkaloid from the acetone-insoluble fraction of the strong bases of this plants. This alkaloid $(C_{25}H_{38}ON_2)$ was identified as a steroidal alkaloid contained a cyclopropane ring by physical and chemical methods. It is a derivative of cyclobuxine D and named cyclobuxine E. We examined the effect of cyclobuxine E on the contractile response induced by acetylcholine and two distinct types of potassium-activated calcium channels in an intestinal smooth muscle of the rat. Cyclobuxine E inhibited significantly the Ach-induced contraction. The isolated longitudinal muscle from the rat duodenum was immersed calcium-depleted potassium depolarizing solution. Ten minutes after, 1.8 mM $CaCl_2$ was added to muscle bath and elicited a biphasic increase in muscle tension. Cyclobuxine E produced an appreciable inhibition of both components of the mechanical response. In addition, Cyclobuxine E introduced at a point when the tonic response had reached its maximum level, caused the muscle to exhibit a rapid loss of tension. Based on these experimental results, we proposed the possibility that the inhibitory action of cyclobuxine E on the isolated rat duodenum may be due to inhibiting the transmembrane fluxes of calcium ion in potassium-activated calcium channels.

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Study on the soluble exoression of recombinant human eoidermal growth factor using various fusion oartners in Escherichia coli (재조합 대장균에서 다양한 융합 파트너를 이용한 인간 상피세포성장인자의 발현 연구)

  • Kim, Byung-Lip;Baek, Jung-Eun;Kim, Chun-Sug;Lee, Hyeok-Weon;Ahn, Jung-Oh;Lee, Hong-Weon;Jung, Joon-Ki;Lee, Eun-Gyo;Kim, In-Ho
    • KSBB Journal
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    • v.23 no.3
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    • pp.205-212
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    • 2008
  • The efficient soluble expression of human epidermal growth factor (hEGF) was achieved by using functional fusion partners in cytoplasm and periplasm of Escherichia coli (E. coli). hEGF was over-expressed in inactive inclusion body form in cytoplasm of E. coli due to improper disulfide bond formation and hydrophobic interaction, yielding about 5.9 mg/L in flask culture. Six functional fusion partners were introduced by linking to N-terminal part of hEGF gene for the high-level expression of soluble and active hEGF in cytoplasm and peri plasm region. Three fusion partners for cytoplasmic expression such as acidic tail of synuclein (ATS), thioredoxin (Trx) and lipase, and three fusion partners for periplasmic expression such as periplasmic cystein oxidoreductases (DsbA and DsbC) and maltose binding protein (MBP) were investigated. hEGF fused with ATS and DsbA showed over 90% of solubility in cytoplasm and periplasm, respectively. Especially DsbA was found to be an efficient fusion partner for soluble and high-level expression of hEGF, yielding about 18.1 mg/L and three-fold higher level compared to that of insoluble non-fusion hEGF in cytoplasm. Thus, heterologous proteins containing complex disulfide bond and many hydrophobic amino acids can effectively be produced as an active form in E. coli by introducing a suitable peptide or protein.

Isolation of Polysaccharides Modulating Mouse’s Intestinal Immune System from Peels of Citrus unshiu (귤피로부터 분리한 마우스의 장관면역 활성 다당류의 검색)

  • Yang, Hyun-Seuk;Yu, Kwang-Won;Choi, Yang-Mun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.9
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    • pp.1476-1485
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    • 2004
  • Of solvent-extracts prepared from the 90 kinds of Korean traditional tea and rice gruel plants, cold-water extract from peels of Citrus unshiu (CUI-0) showed the most potent intestinal immune system modulating activity through Peyer’s patch whereas other extracts did not have the activity except for cold-water extracts of Laminaria japonica, Polygonatum japonicum, Poncirus trifoliata, and hot-water extracts of Gardenia jasminoides, Lycium chinense having intermediate activity. CUI-0 was further fractionated into MeOH-soluble fraction (CUI-1), MeOH insoluble and EtOH-soluble fraction (CUI-2), and crude polysaccharide fraction (CUI-3). Among these fractions, CUI-3 showed the most potent stimulating activity for the proliferation of bone marrow cells mediated by Peyer’s patch cells, and contained arabinose, galacturonic acid, galactose, glucose, glucuronic acid and rhamnose (molar ratio; 1.00:0.53:0.45:0.28:0.28:0.19) as the major sugars, and a small quantity of protein (9.4%). In treatments of CUI-3 with pronase and periodate (NaIO₄), the intestinal immune system modulating activity of CUI-3 was significantly reduced, and the activity of CUI-3 was affected by periodate oxidation particularly. The potently active carbohydrate-rich fraction, CUI-3IIb-3-2 was further purified by anion-exchange chromatography on DEAE-Sepharose FF, Sepharose CL-6B and Sephacryl S-200. CUI-3IIb-3-2 was eluted as a single peak on HPLC and its molecular weight was estimated to be 18,000 Da. CUI-3IIb-3-2 was consisted mainly of arabinose, galactose, rhamnose, galacturonic acid and glucuronic acid (molar ratio;1.00:0.54:0.28:1.45:0.63) in addition to a small amount of proteins (3.2%). In addition, CUI-3IIb-3-2 showed the activity only through Peyer’s patch cells, but this fraction did not directly stimulate proliferation of bone marrow cells. It may be concluded that intestinal immune system modulating activity of peels from C. unshiu is caused by pectic polysaccharides having a polygalacturonan moiety with neutral sugars such as arabinose and galactose.

Growth of Landscape Tree Species at Two Planting Densities in a Planting Pilot System for Reclaimed Dredging Areas (임해준설매립지 식물재배공정에서 밀도에 따른 조경수목의 생장)

  • Lee, Deok-Beom;Nam, Woong;Kwak, Young-Se;Jeong, In-Ho;Lee, Sang-Suk
    • Journal of the Korean Institute of Landscape Architecture
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    • v.37 no.2
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    • pp.114-123
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    • 2009
  • To investigate the possible use of plants for landscaping in reclaimed soil, a planting pilot system experiment was performed over the course of four years in reclaimed dredging area with four species: Alnus firma, Alnus hirsuta, Pinus thunbergii, and Pyrachantha angustifolia for 4 years. The physicochemical characteristics of the tested soil showed that it was sandy through coming from a reclaimed dredging area. The average pH of the tested soil was 7.16(slight alkali), and electric conductivity(EC) was relatively low, $294{\mu}S/cm$, even though it came from a saltwater area. To test the effect of planting density vs. phytomass by plant specie from a planting basin, the experiment was designed using four plant species with high and low planting densities over 4 years. The planting conditions of the growth of landscape tree species exhibited growth height as follows: A. hirsuta, A. firma, P. thunbergii, and P. angustifolia, whill the DBH followed the order of A. hirsuta, A. firma, and P. thunbergii. The total phytomass of each plant was higher at low density planting areas than high density planting area in terms of total phytomass production and growth distribution in the reclaimed dredging area. Total phytomass per unit area increased as follows: A. hirsuta, A. firma, P. thunbergii, and P. angustifolia. The total phytomass per each tested plant was 2 times higher in low density planting areas than high density planting areas. Total phytomass per unit area, however, was similar or slighty higher in high density planting areas compared to low density areas. Among the tested plants, A. hirsuta showed the highest phytomass, implying that A. hirsuta adapted very well to the reclaimed area and has the capability of a fast growth, nitrogen fixation tree, and utilizing insoluble nutrients through inoculated root nodule bacteria. The yield of phytomass per individual in low density Alnus species was greater than that of the high density. However, those per unit areas had no difference in the density-dependent planting. The ratio of belowground to aboveground was $0.21{\sim}0.26$. Thus, it could be concluded that the Alnus species are potential candidates for ornamental tree species in reclaimed dredging areas. This study offers baseline data for the use of ornamental tree species in reclaimed dredging areas. Additional research is required for different ornamental species in order to increase phytomass of a planting conditions based on reclaimed dredging areas.

A Study on Public Nuisance in Kwangju City (Survey on Air Pollution and Noise Level) (공해(公害)에 관(關)한 조사연구(調査硏究) -광주시(光州市)의 대기오염(大氣汚染) 및 소음(騷音)에 관(關)한 조사연구(調査硏究)-)

  • Chung, Yo-Han;Kim, Kil-Wng;Moon, Jae-Kyu;Jhoo, Heung-Kyu
    • Journal of Preventive Medicine and Public Health
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    • v.5 no.1
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    • pp.1-7
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    • 1972
  • During the period from June 1st 1971 to November 30 th 1971, studies on air pollution were made in Kwangju city. The city was divided into 6 areas; the downtown area, the semi-downtown area, the heavy traffic area, the commercial area, the residential area, the park area, 13 surveying sites were selected each representing the characteristics of the area. The Measurement methods which were used are described below. Sulfur oxides were measured by $PbO_2$ cylinder method, sulfur dioxides ($SO_2$) and carbon monoxide (CO) by the MSA & Kitakwa detector, dustfall by the Deposit gauge method, and the noise levels by the Kanomax soundlevel meter. The results obtained are as follows: 1. The mean value of sulfur oxides in Kwangju city was $1.16mg\;SO_3/day/100cm^2\;PbO_2$, ranging from $0.45mg\;SO_3/day/100cm^2\;PbO_2$ to $3.10mg\;SO_3/day/100cm^2\;PbO_2$. 2. The mean values of sulfur oxides according to its specific area in the city were $1.45mg\;SO_3/day/100cm^2\;PbO_2$ in heavy traffic area, 1.36 in downtown area, 1.23 in semi-downtown area, 1.11 in commercial area, 0.96 in residential area, and 1.07 in park area, respectively. 3. The average concentration of sulfur dioxide was 0.063 ppm from 2 to 5 P.M in Kwangju city. 4. The average concentrations of sulfur dioxides according to its specific area, from 2 to 5 P.M, in the city were 0.084 ppm in heavy traffic area & downtown area, 0.067 in commercial area, 0.053 in semi-downtown area, 0.052 in residential area, and 0.036 in park area. 5. The average concentration of carbon monoxide was 22.3 ppm from 2 to 5 P.M, in Kwangju city. 6. The average concentrations of carbon monoxide according to its specific area, from 2 to 5 P.M, in the city were 27.0 ppm in downtown area, 26.3 in semi-downtown area, 23.0 in heavy traffic area, 21.7 in commercial area, 20.0 in residential arera, and 17.6 in park area. 7. The mean value of dusifall in Kwangju city was $29.28ton/km^2/month$, ranging from $9.85ton/km^2/month$ to $66.34ton/km^2/month$. 8. The mean values of dustfall according to its specific area in the city were $50.37ton/km^2/month$ in semi-downtown area, 42.76 in heavy traffic area, 34.67 in downtown area, 17.77 in commercial area, 14.40 in park area, and 14.76 in residential area. 9. The mean values of the soluble dust in Kwangju city was $10.23ton/Km^2/month$ and that of the insoluble dust was $19.05ton/Km^2/month$. 10. The mean value of noise level in Kwangju city was 62 phon, ranging from 37 phon to 88 phon. 11. The mean values of noise level according to its specific area in the city were 76 phon in heavy traffic area, 67 in semi-downtown area, 64 in downtown area, 59 in commercial area, 52 in part area, and 50 in residential area.

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Toxicity Assessment of Silver Ions Compared to Silver Nanoparticles in Aqueous Solutions and Soils Using Microtox Bioassay (Microtox 생물검정법을 이용한 은 이온과 은 나노입자의 수용액과 토양에서의 독성 비교 평가)

  • Wie, Min-A;Oh, Se-Jin;Kim, Sung-Chul;Kim, Rog-Young;Lee, Sang-Phil;Kim, Won-Il;Yang, Jae E.
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.6
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    • pp.1114-1119
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    • 2012
  • This study was conducted to assess the microbial toxicity of ionic silver solution ($Ag^+N$) and silver nanoparticle suspension ($Ag^0NP$) based on the Microtox bioassay. In this test, the light inhibition of luminescent bacteria was measured after 15 and 30 min exposure to aqueous solutions and soils spiked with a dilution series of $Ag^+N$ and $Ag^0NP$. The resulting dose-response curves were used to derive effective concentration (EC25, $EC_{50}$, EC75) and effective dose ($ED_{25}$, $ED_{50}$, $ED_{75}$) that caused a 25, 50 and 75% inhibition of luminescence. In aqueous solutions, $EC_{50}$ value of $Ag^+N$ after 15 min exposure was determined to be < $2mg\;L^{-1}$ and remarkably lower than $EC_{50}$ value of $Ag^0NP$ with $251mg\;L^{-1}$. This revealed that $Ag^+N$ was more toxic to luminescent bacteria than $Ag^0NP$. In soil extracts, however, $ED_{50}$ value of $Ag^+N$ with 196 mg kg-1 was higher than $ED_{50}$ value of $Ag^0NP$ with $104mg\;kg^{-1}$, indicating less toxicity of $Ag^+N$ in soils. The reduced toxicity of $Ag^+N$ in soils can be attributed to a partial adsorption of ionic $Ag^+$ on soil colloids and humic acid as well as a partial formation of insoluble AgCl with NaCl of Microtox diluent. This resulted in lower concentration of active Ag in soil extracts obtained after 1 hour shaking with $Ag^+N$ than that spiked with $Ag^0NP$. With longer exposure time, EC and ED values of both $Ag^+N$ and $Ag^0NP$ decreased, so their toxicity increased. The toxic characteristics of silver nanomaterials were different depending on existing form of Ag ($Ag^+$, $Ag^0$), reaction medium (aqueous solution, soil), and exposure time.

Establishment of an Ice Core Processing Method and Analytical Procedures for Fundamental Proxies (빙하코어의 전처리 방법 및 기초 프록시 분석법 확립)

  • Jun, Seong Joon;Hong, Sang Bum;Hur, Soon Do;Lee, Jeonghoon;Kang, Jung-Ho;Hwang, Hee Jin;Chung, Ji Woong;Jung, Hye Jin;Han, Changhee;Hong, Sungmin
    • Ocean and Polar Research
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    • v.36 no.1
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    • pp.13-24
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    • 2014
  • We established the first complete ice core processing method and analytical procedures for fundamental proxies, using a 40.2 m long ice core drilled on the Mt. Tsambagarav glacier in the Mongolian Altai mountains in July 2008. The whole core was first divided into two sub ice core sections and the measurements of the visual stratigraphy and electrical conductivity were performed on the surface of these sub core sections. A continuous sequence of samples was then prepared for chemical analyses (stable isotope ratios of oxygen ($^{18}O/^{16}O$) and hydrogen ($^2H/^1H$), soluble ions and trace elements). A total of 29 insoluble dust layers were identified from the measurement of visual stratigraphy. The electrical conductivity measurement (ECM) shows 11 peaks with the current more than 0.8 ${\mu}A$ Comparing the profiles of $SO_4{^{2-}}$ and $Cl^-$ concentrations to correlate with known volcanic eruptions, the first two ECM peaks appear to be linked to the eruptions (January and June 2007) of Kliuchevskoi volcano on the Kamchatka Peninsula of Russia, which supports the reliability of our ECM data. Finally, the composition of stable isotopes (${\delta}^{18}O$ and ${\delta}D$) shows a well-defined seasonal variation, suggesting that various chemical proxies may have been well preserved in the successive ice layers of Tsambagarav ice core. Our ice core processing method and analytical procedures for fundamental proxies are expected to be used for paleoclimate and paleoenvironmental studies from polar and alpine ice cores.

Organic Matter Analysis and Physicochemical Properties of Leachate from a Foot-and-Mouth Disease Landfill Site (구제역 가축 매몰지 침출수의 물리 화학적특성과 유기물질 성상분석)

  • Kang, Mee-A;Kim, Mi-Sun;Choi, Byung-Woo;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.128-134
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    • 2012
  • Foot and mouth disease (FMD) is one of the most notorious and contagious viral diseases afflicting cloven-hoofed animals. In this study, the physicochemical properties of leachate from a FMD landfill site at 773-1, Waryong, Andong, Korea and the ground water from 777, Waryong, Andong, Korea, were analyzed for 1 year from December $10^{th}$ 2010 to November $17^{th}$ 2011. The leachate was collected from the FMD landfill site during March, May, July, September and November, 2011 and changes in pH, brix, water content, insoluble solids, crude proteins, crude lipids, total and reducing sugars and ash content were determined. Considering the annual profiles of temperature and rainfall at the FMD landfill site, the dramatic changes in the physicochemical properties of the leachate from March to July, and especially from May to July, such as increases in pH, and a rapid reduction of brix and organic matter, may be closely linked to the growth of microorganisms in the leachate. The sharp decreases in the concentration of biominerals, such as Mg, Ca, and Fe from 1073, 4311 and 56.2 ppm in March to 151, 78, and 0.1 ppm in November, further suggest that decreases in organic matter in the leachate result from degradation by microorganisms originating from the intestines of the livestock. Analysis of the profiles of the organic materials in the leachate revealed that the properties of the leachate were similar to those of excremental matter-derived water. These results could be applied to a number of fields for the analysis of organic matter behavior, the development of the degradation process, and risk analysis in the environment for hygiene and food industries, of leachate from FMD landfill sites.

Sorption of PAHs by Soil Humins and Effect of Soil Inorganic Matrixs (PAHs의 토양휴민과의 흡착특성 및 토양 무기물의 영향 해석)

  • Lim, Dong-Min;Lee, Seung-Sik;Shin, Hyun-Sang
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.12
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    • pp.1337-1346
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    • 2006
  • Soil humin is the insoluble fraction of humic materials and play an important roles in the irreversible sorption of hydrophobic organic contaminants onto soil particles. However, there have been limited knowledge about the sorption and chemical properties of humin due to the difficulties in its separation from the inorganic matrix(mainly clays and oxides). In this study, de-ashed soil humins($Hu_1-Hu_6$) were isolated from a soil residues(Crude Hu) after removing alkali-soluble organic fractions followed by consecutive dissolution of the mineral matrix with 2%-HF for 2 hr. The humin samples were characterized by elemental analysis and $^{13}C$ NMR spectroscopic method and their sorption-desorption behavior for 1-naphthol were investigated from aqueous solution. The results were compared one another and that with peat humin. $^{13}C$ NMR spectra features indicate that the soil humin molecules are mainly made up of aliphatic carbons(>80% in total carbon) including carbohydrate, methylene chain. Freundlich sorption parameter, n was increased from 0.538 to 0.697 and organic carbon-normalized sorption coefficient(log $K_{OC}$) values also increased from 2.43 to 2.74 as inorganic matrix of the soil humin removed by HF de-ashing. The results suggest that inorganic phase in humin plays an important, indirect role in 1-naphthol sorption and the effects on the sorption non-linearity and intensity are analyzed by comparison between the results of soil humin and peat humin. Sorption-desorption hysteresis were also observed in all the humin samples and hysteresis index(HI) at low solute concentration($C_e$=0.1 mg/L) are in order of Peat humin(2.67)>De-ashed humin(0.74)>Crude Hu(0.59).