• Title/Summary/Keyword: base-isolated

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In Vitro Differentiation of Human Amniotic Membrane-derived Stem Cells into Hepatocyte-like Cells (양막 유래 줄기세포의 간세포로의 분화 유도)

  • Kook, Min-Ji;Park, Soo-Yeon;Kang, Hyun-Mi;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.10 no.1
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    • pp.63-73
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    • 2006
  • This study aimed to find out suitable culture conditions for the differentiation of human amniotic membrane-derived stem cells(HAM) into hepatocyte-like cells. Almost homogenous population of fibroblast-like cells was successfully isolated from the amniotic membrane. In comparison to the non-coated plates and in the absence of insulin/transferrin/selenium(ITS), HAM cultured on the fibronectin-coated plates and in the presence of ITS showed the more intense immunocytochemical staining against the albumin. Addition of both fibroblast growth factor(FGF)-1 and -2 to the differentiation medium gave stronger staining compared to the treatment with FGF-1 or -2 alone. Periodic acid Schiff's base staining of glycogen and morphological turnover of fibroblast-like appearance of HAM into round shape matched the results of immunocytochemical studies. When the efficiency of two-step culture method was examined on the differentiation of HAM into hepatocyte-like cells, all of the results of immunocytochemical staining, periodic acid Schiff's staining and morphological change exhibited effective hepatic differentiation of HAM compared to the continuous culture method. Immunoblot analyses of HAM- conditioned media against the albumin showed that the culture of HAM in the presence of both ITS and fibronectin always gave a stronger staining intensity than those in the absence of them, and that the addition of ether mixture of FGF-4 and either FGF-2 or transforming growth $factor(TGF)-{\alpha}$ to the culture medium significantly enhanced the albumin secretion by HAM. Based on these observations, it is suggested that HAM could differentiate into hepatocyte-like cells under a culture condition consisting of fibronectin and ITS, and addition of FGF-4 with either one of FGF-2 or $TGF-{\alpha}$ could enhance the hepatic differentiation of HAM.

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Effect of Silicate-Coated Rice Seed on Healthy Seedling Development and Bakanae Disease Reduction when Raising Rice in Seed Boxes (벼 상자육묘에서 규산코팅볍씨의 건묘육성과 벼키다리병 경감효과)

  • Kang, Yang-Soon;Kim, Wan Jung;Roh, Jae-Hwan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.62 no.1
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    • pp.1-8
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    • 2017
  • We investigated the effect of silicate coating of rice seeds on bakanae disease incidence and the quality of seedlings raised in seedling boxes and transplanted into pots. The silicate-coated rice seed (SCS) was prepared as follows. Naturally infested rice seeds not previously subjected to any fungicidal treatment were dressed with a mixture of 25% silicic acid at pH 11 and 300-mesh zeolite powder at a ratio of 50 g dry seed - 9 mL silicic acid - 25 g zeolite powder. The following nursery conditions were provided : Early sowing, dense seeding in a glass house with mulching overnight and no artificial heating, which were the ideal conditions for determining the effect on the seed. The nursery plants were evaluated for Gibberella. fujikuroi infection or to determine the recovery to normal growth of infected nursery plants in the Wagner pot. Seedlings emerged 2-3 days earlier for the SCS than they did for the non-SCS control, while damping-off and bakanae disease incidence were remarkably reduced. Specifically, bakanae disease incidence in the SCS was limited to only 7.8% for 80 days after sowing, as compared to 91.6% of the non-SCS control. For the 45-days-old SCS nursery seedlings, the fresh weight was increased by 11% and was two times heavier, with only mild damage compared to that observed for non-SCS. Even after transplanting, SCS treatment contributed to a lower incidence of further infections and possibly to recovery of the seedlings to normal growth as compared to that observed in symptomatic plants in the pot. The active pathogenic macro-conidia and micro-conidia were considerably lower in the soil, root, and seedling sheath base of the SCS. In particular, the underdeveloped macro-conidia with straight oblong shape without intact septum were isolated in the SCS ; this phenotype is likely to be at a comparative etiological disadvantage when compared to that of typical active macro-conidia, which are slightly sickle-shaped with 3-7 intact septa. A active intact conidia with high inoculum potential were rarely observed in the tissue of the seedlings treated only in the SCS. We propose that promising result was likely achieved via inhibition of the development of intact pathogenic conidia, in concert with the aerobic, acidic conditions induced by the physiochemical characteristics associated with the air porosity of zeolite, alkalinity of silicate and the seed husk as a carbon source. In addition, the resistance of the healthy plants to pathogenic conidia was also important factor.

Genetic Differences and Variation in Two Largehead Hairtail (Trichiurus lepturus) Populations Determined by RAPD-PCR Analysis (RAPD-PCR 분석에 의해 결정된 갈치 (Trichiurus lepturus) 2 집단의 유전적 차이와 변이)

  • Park, Chang-Yi;Yoon, Jong-Man
    • Korean Journal of Ichthyology
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    • v.17 no.3
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    • pp.173-186
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    • 2005
  • Genomic DNA was isolated from two geographic populations of largehead hairtail (Trichiurus lepturus) in Korea and the Atlantic Ocean. The eight arbitrarily selected primers were found to generate common, polymorphic, and specific fragments. The complexity of the banding patterns varied dramatically between primers from the two locations. The size of the DNA fragments also varied widely, from 150 bp (base pairs) to 3,000 bp. Here, 947 fragments were identified in the largehead hairtail population from Korea, and 642 in the largehead hairtail population from the Atlantic Ocean: 148 specific fragments (15.6%) in the Korean population, and 61 (9.5%) in the Atlantic population. In the Korean population, 638 common fragments with an average of 79.8 per primer were observed.; 429 common fragments, with an average of 53.6 per primer, were identified in the Atlantic population. The number of polymorphic fragments in the largehead hairtail population from Korea and the Atlantic Ocean was 76 and 27, respectively. Based on the average bandsharing values of all samples, the similarity matrix ranged from 0.784 to 0.922 in the Korean population, and from 0.833 to 0.990 in the Atlantic population. The bandsharing value of individuals within the Atlantic population was much higher than in the Korean population. The dendrogram obtained by the eight primers indicated two genetic clusters: cluster 1 (KOREAN 01~KOREAN 11), and cluster 2 (ATLANTIC 12~ATLANTIC 22). Individual KOREAN no. 10 from Korea was genetically most closely related to KOREAN no. 11 in the Korean population (genetic distance = 0.038). Ultimately, individual KOREAN no. 01 of the Korean population was most distantly related to ATLANTIC no. 16 of the Atlantic population (genetic distance = 0.708).

Current status, mechanism and control of herbicide resistant weeds in rice fields of Korea (한국 논에서 제초제 저항성잡초의 발생 현황, 메카니즘 및 방제)

  • Park, Tae Seon;Seong, Ki Yeong;Cho, Hyun Suk;Seo, Myung Chul;Kang, Hang Won;Park, Kee Woong
    • Korean Journal of Agricultural Science
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    • v.41 no.2
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    • pp.85-99
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    • 2014
  • Sulfonylurea (SU)-resistant weeds of eight annual weeds, Monochoria vaginalis, Scirpus juncoides and Cyperus difformis, etc., and four perennial weeds, Scirpus planiculmis, Sagittaria pigmaea, Eleocharis acicularis and Sagittaria trifolia as of 2013 since identification Monochoria korsakowii in the reclaimed rice field in 1998. And the resistant Echinochloa oryzoides to ACCase and ALS inhibitors has been confirmed in rice fields of the southern province, Korea in 2009. In the beginning, the M. vaginalis, S. juncoides and C. difformis of these SU-resistant weeds were rapidly and individually spreaded in different fields, however, these resistant weeds have been occurring simultaneously in the same filed recently. The resistant biotype by weed species demonstrated about 10-to 1,000-fold resistance, based on $GR_{50}$ values of the SU herbicides tested. And the resistant biotype of E. oryzoides to cyhalofop-butyl, pyriminobac-methyl, and penoxsulam was about 14, 8, and 11 times more resistant than the susceptible biotype base on $GR_{50}$ values. The products and applied area of SU-included herbicides have been increased rapidly, and have accounted for about 69% and 96% in Korea, respectively. In Korea, the main cause of SU-resistant weed is extensive use of these herbicides. The top ten herbicides by applied area were composed of all SU-included herbicides by 2003. The concentrated and successive treatment of ACCase and ALS inhibitors for control of barnyardgrass in rice led up to the resistance of E. oryzoides. Also, SU-herbicides like pyrazosulfuron-ethyl and imazosulfuron which effective to barnyardgrass can be bound up with the resistance of E. oryzoides. The ALS activity isolated from the resistant biotype of M. korsakowii to SU-herbicides tested was less sensitive than that of susceptible biotype. The concentration of herbicide required for 50% inhibition of ALS activity ($I_{50}$) of the SU-resistant M. korsakowii was 14-to 76-fold higher as compared to the susceptible biotype. No differences were observed in the rates of [$^{14}C$]bensulfuron uptake and translocation. Acetolactate synthase (ALS) genes from M. vaginalis resistant and susceptible biotypes against SU-herbicides revealed a single amino acid substitution of proline (CCT), at 197th position based on the M. korsakowii ALS sequence numbering, to serin (TCT) in conserved domain A of the gene. Carfentrazone-ethyl and pyrazolate were used mainly to control SU-resistant M. vaginalis by 2006 in Korea. However, the alternative herbicides such as benzobicyclone, to be possible to control simultaneously the several resistant weeds, have been developing and using broadly, because the several resistant weeds have been occurring simultaneously in the same fieled. The top ten herbicides by applied area in Korea have been occupied by products of 3-way mixture type including herbicides with alternative mode of action for the herbicide resistant weeds. Mefenacet, fentrazamide and cafenstrole had excellent controlling effects on the ACCase and ALS inhibitors resistant when they were applied within 2nd leaf stage.

THE TASTE COMPOUNDS FERMENTED ACETES CHINENSIS (새우젓의 정미성분에 관한 연구)

  • CHUNG Seung-Yong;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.9 no.2
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    • pp.79-110
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    • 1976
  • In Korea fermented fish and shellfish have traditionally been favored and consumed as seasonings or further processed for fish sauce. Three major items in production quantity among more than thirty kinds which are presently available in the market are fermented anchovy, oyster and small shrimp. They are usually used as a seasoning mixture of Kimchi in order to provide a distinctive flavor. Fermented small shrimp, Acetes chinensis is most widely and largely used ana occupies an important position in food industry of this country. But no study on its taste compounds has been reported. This study was attempted to establish the basic data for evaluating taste compounds of fermented small shrimp. The changes of such compounds during fermentation as free amino acids, nucleotides and their related compounds, TMAO, TMA, and betaine were analysed. In addition, change in microflora during the fermentation under the halophilic circumstance was also investigated. The samples were prepared with three different salt contents of 20, 30 and $40\%$ to obtain the proper degree of fermentation at a controlled tempeature of $20{\pm}2^{\circ}C$. The results are summarized as follows: Volatile basic nitrogen increased rapidly until 108 days of fermentation and afterwards it tended to increase slowly. Amino nitrogen also increased rapidly until 43 days of fermentation and then increased slowly. Extract nitrogen increased and marked the maximum value at 72 day fermentation and then decreased slowly. ADP, AMP and IMP tended to degrade rapidly while hypoxanthine increased remarkably at 27 day fermentation but slightly decreased at 72 day fermentation. It is presumed that the characteristic flavor of fermented small shrimp might be attributed to the relatively higher content of hypoxanthine. In the free amino acid composition of fresh small shrimp abundant amino acids were proline, arginine, alanine, glycine, lysine, glutamic acid, leucine, valine and threonine in order. Such amino acids like serine, methionine, isoleucine, phenylalanine, aspartic acid, tyrosine and histidine were poor. In small shrimp extract, proline, arginine, alanine, glycine, lysine and glutamic acid were dominant holding $18.5\%,\;14.6\%,\;10.8\%,\;8.7\%,\;8.1\%\;and\;7.7\%$ of total free amino acids respectively. The total free amino acid nitrogen in fresh small shrimp was $63.9\%$ of its extract nitrogen. The change of free amino acid composition in the extract of small shrimp during fermentation was not observed. Lysine, alanine glutamic acid, proline, glycine and leucine were abundant in both fresh sample and fermented products. The increase of total free amino acids during 72 day fermentation reached approximately more than 2 times as compared with that of fresh sample and then decreased slowly. Fermented small shrimp with $40\%$ of salt was too salty to be commercial quality as the results of organoleptic test showed. It is found that 72 day fermentation with $20\%\;and\;30\%$ of salt gave the most favorable flavor. It is convinced that the characteristic flavor of fermented small shrimp was also attributed to such amino acids as lysine, proline, alanine, glycine and serine known as sweet compounds, as glutamic acid with meaty taste, and as leucine known as bitter taste. The amount of betaine increased during fermentation and reached the maximum at 72 day fermentation and then decreased slowly TMA increased while TMAO decreased during fermentation. The amount of TMAO nitrogen in fermented small shrimp was $200mg\%$ on moisture and salt free base. Betaine and TMAO known as sweet compounds were abundant in fermented small shrimp. It is supposed that these compounds could also play a role as important taste compounds of fermented small shrimp. At the initial stage of fermentation, Achromobacter, Pseudomonas, Micrococcus denitrificans which belong to marine bacteria were isolated. After 40 day fermentation, they disappeared rapidly while Halabacterium, Pediococcus, Sarcian, Micrococcus morrhuae and the yeasts such as Saccharomyces sp. and Torulopsis sp. dominated. It is concluded that the most important taste compounds of fermented small shrimp were amino acids such as lysine, proline, alanine, glycine, serine, glutamic acid, and leucine, betaine, TMAO and hypoxanthine.

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A Study on Safety of Ready-to-eat Compound Foods with a By-products of Meat as the Base (식육부산물을 주재료로한 복합즉석조리식품의 안전성 연구)

  • Song, Sung-Min;Lee, Gil-Bong;Kim, Myeong-Hee;Jeung, Ji-Yeol;Hwang, Won-Mu;Yun, Ga-Ri;Kim, Sun-Hoi;Go, Jong-Myeung;Kim, Yong-Hee
    • Journal of Food Hygiene and Safety
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    • v.22 no.2
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    • pp.82-87
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    • 2007
  • This research was performed to investigate chemical and microbiological hazards of Ready-to-eat(RTE) compound foods which uses the by-product of meat. For this experiment, 51 samples of RTE compound foods in by-product of meat distributed in Incheon from January to December 2006 were tested. The contents of heavy metals in the main ingredient of RTE compound foods were in the range of $0.073{\sim}0.112ppm$ for lead (Pb), $0.006{\sim}0.013ppm$ for cadmium (Cd) and $0.746{\sim}0.978ppb$ for mercury (Hg). The concentrations of residual ABS(alkyl benzene sulfate) in the small intestine which is a main ingredient of Gopchang-casserole were $$0.8ppm{\sim}57.6ppm$ (Ave. 10.3ppm). Staphylococcus aureus was isolated from 11 samples (21.6%) among 51 main ingredients of RTE compound foods. The isolation rates of Salmonella spp. and Clostridium perfringens were 2.0% (1/51) and 5.9% (2/51), respectively. By types of main ingredient, the small intestine was showed the highest isolation rate as 35.3% (12/34), ham and the sausage which are main ingredients of the Budae-pot stew were 25% (2/8) and other meat products were 20% (1/5). Food poisoning bacteria was not found in the blood of pig which is a main ingredient of the Sunji-pot stew. 28.4% (27/95) of sauce included in each RTE compound foods were coliform bacteria positive. Pesticide residues were found in four of 45 vegetables which are the additional ingredient of RTE compound foods. The concentrations of pesticide were chlorothalonil 2.8 ppm, EPN 10.3 ppm, chlorpyrifos 0.4ppm and indoxacarb 0.7ppm. In 33 bean sprout samples, captan and carbendazim were not detected.

Cis-acting Replication Element Variation of the Foot-and-mouth Disease Virus is Associated with the Determination of Host Susceptibility (구제역바이러스의 숙주 특이성 결정에 연관되어있는 구제역바이러스 cis-acting replication element 변이 분석 연구)

  • Kang, Hyo Rin;Seong, Mi So;Ku, Bok Kyung;Cheong, JaeHun
    • Journal of Life Science
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    • v.30 no.11
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    • pp.947-955
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    • 2020
  • The foot-and-mouth disease virus (FMDV), a member of the Aphthovirus genus in the Picornaviridae family, affects wild and domesticated ruminants and pigs. During replication of the FMDV RNA (ribonucleic acid) genome, FMDV-encoding RNA polymerase 3D acts in a highly location-specific manner. This suggests that specific RNA structures recognized by 3D polymerase within non-coding regions of the FMDV genome assist with binding during replication. One such region is the cis-acting replication element (CRE), which functions as a template for RNA replication. The FMDV CRE adopts a stem-loop conformation with an extended duplex stem, supporting a novel 15-17 nucleotide loop that derives stability from base-stacking interactions, with the exact RNA nucleotide sequence of the CRE producing different RNA secondary structures. Here, we show that CRE sequences of FMDVs isolated in Korea from 2010 to 2017 exhibit A and O genotypes. Interestingly, variations in the RNA secondary structure of the Korean FMDVs are consistent with the phylogenetic relationships between these viruses and reveal the specificity of FMDV infections for particular host species. Therefore, we conclude that each genetic clade of Korean FMDV is characterized by a unique functional CRE and that the evolutionary success of new genetic lineages may be associated with the invention of a novel CRE motif. Therefore, we propose that the specific RNA structure of a CRE is an additional criterion for FMDV classification dependent on the host species. These findings will help correctly analyze CRE sequences and indicate the specificity of host species for future FMDV epidemics.

Analysis of rpoB Gene in Rifampin-Resistant M. Tuberculosis by Direct Sequencing and Line Probe Assay (염기서열결정과 Line Probe 분석법에 의한 Rifampin내성 결핵균의 rpoB 유전자 분석)

  • Lee, Min-Ki;Kim, Yun-Seong;Lee, Hyo-Jin;Cheon, Du-Su;Yun, Sang-Myung;Park, Sam-Seok;Kim, Cheol-Min;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.2
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    • pp.251-263
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    • 1997
  • Background : The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture. Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in $\beta$subunit of RNA polymerase is main mechanism of resistance. Method : In this study, rpoB gene for the $\beta$subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. Results : All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S531L) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not confirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513~515 were new identified mutations for the first time. Conclusion : Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.

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Expression of Anion Exchanger and CFTR in the Hepatocyte and Cholangiocytes in Bile Duct-Ligated Rat (담관 결찰 쥐의 간세포와 담관세포의 anion exchanger와 CFTR 발현)

  • Lee, Jae-Dong;Wang, Joon-Ho;Ki, Seung-Seog;Choe, Won-Hyeok;Park, Jae-Seung;Cho, Won-Kyoo;Park, Jung-Jun;Kim, Hong-Su
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1772-1777
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    • 2011
  • Cystic fibrosis transmembrane conductance regulator (CFTR) gene and sodium-independent $Cl^-/HCO_3^-$ anion exchanger (AE) genes are expressed in a wide variety of mammalian tissues including cholangiocytes. They play an important role in the regulation of intracellular pH (pHi) as well as in transepithelial acid/base transport necessary for biliary bicarbonate secretion. The aim of this study was to examine the expression level of CFTR gene and AE genes (AE1, AE2 and AE3) in the cholangiocytes and the hepatocytes, and also measure AE2 gene expression level after bile duct ligation (BDL). As we previously described, isolated hepatocytes and cholangiocytes from the liver of normal and BDL rats were prepared and gene expression levels were measured by using RT-PCR. We found that AE1, AE2, and AE3 genes were expressed in both hepatocytes and cholangiocytes, but CFTR was only in cholangiocytes. AE2 gene expression level was higher in the BDL hepatocytes than normal hepatocytes, which was significantly different between two groups. AE2 gene expression level was lower in the BDL cholangiocytes than normal cholangiocytes. However, AE2 gene expression level in both hepatocytes and cholangiocytes were not changed with a longer duration of BDL. These results suggest that CFTR and AE2 may play an important role in the pathogenetic mechanism of biliary cholestatic liver disease.

Chemical and Spectroscopic Characterization of Peat Moss and Its Different Humic Fractions (Humin, Humic Acid and Fulvic Acid) (피트모스에서 추출한 휴믹물질(휴믹산, 풀빅산, 휴민)의 화학적 및 분광학적 물질특성 규명)

  • Lee Chang-Hoon;Shin Hyun-Sang;Kang Ki-Hoon
    • Journal of Soil and Groundwater Environment
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    • v.9 no.4
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    • pp.42-51
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    • 2004
  • Peat humin(p-Humin), humic acid(p-HA) and fulvic acid(p-FA) were isolated from Canadian Sphagnum peat moss by dissolution in 0.1M NaOH followed by acid precipitation. After purification cycles, they are characterized for their elemental compositions and, acid/base properties. Functionalities and carbon structures of the humic fractions were also characterized using FT-IR and solid state $^{13}C$-NMR spectroscopy. Those results are compared with one another and with soil humic substances from literatures. Main purpose of this study was to present a chemical and spectroscopic characterization data of humic substance from peat moss needed to evaluate its environmental applicability. The relative proportions of the p-Humin, p-HA and p-FA in the peat moss was $76\%,\;18\%,\;and\;3\%$, respectively, based on the total organic matter content ($957{\pm}32\;g/kg$). Elemental composition of p-Humin were found to be $C_{1.00}H_{1.52}O_{0.79}N_{0.01}$ and had higher H/C and (N+O)/C ratio compared to those of p-HA($C_{1.00}H_{1.09}O_{0.51}N_{0.02}$) and p-FA($C_{1.00}H_{1.08}O_{0.65}N_{0.01}$). Based on the analysis of pH titration data, there are two different types of acidic functional groups in the peat moss and its humic fractions and their proton exchange capacities(PEC, meq/g) were in the order p-FA(4.91) >p-HA(4.09) >p-Humin(2.38). IR spectroscopic results showed that the functionalities of the peat moss humic molecules are similar to those of soil humic substances, and carboxylic acid(-COOH) is main function group providing metal binding sites for Cd(II) sorption. Spectral features obtained from $^{13}C$-NMR indicated that peat moss humic molecules have rather lower degree of humification, and that important structural differences exist between p-Humin and soluble humic fractions(p-HA and p-FA).