• Title/Summary/Keyword: $NH_3-NH_4^+$

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Bacillus subtilis를 이용한 대두 발효식품의 혈전용해능

  • Jeong, Yeong-Gi
    • Proceedings of the Korean Society of Life Science Conference
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    • 2001.06a
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    • pp.67-86
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    • 2001
  • A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

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Validation on the Analytical Method of Ginsenosides in Red Ginseng

  • Cho B. G.;Nho K. B.;Shon H. J.;Choi K. J.;Lee S. K.;Kim S. C;Ko S. R.;Xie P. S.;Yan Y. Z.;Yang J. W.
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.491-501
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    • 2002
  • A cross-examination between KT&G Central Research Institute and Guangzhou Institute for Drug Control was carried out in order to select optimum conditions for extraction, separation and determination of ginsenosides in red ginseng and to propose a better method for the quantitative analysis of ginsenosides. The optimum extraction conditions of ginsenosides from red ginseng were as follows: the extraction solvent, $70\%$ methanol; the extraction temperature, $100^{\circ}C;$ the extraction time, 1 hour for once; and the repetition of extraction, twice. The optimum separation conditions of ginsenosides on the SepPak $C_{18}$ cartridge were as follows: the loaded amount, 0.4 g of methanol extract; the washing solvents, distilled water of 25 ml at first and then $30\%$ methanol of 25 ml; the elution solvent, $90\%$ methanol of 5 ml. The optimum HPLC conditions for the determination of ginsenosides were as follows: column, Lichrosorb $NH_2(25{\times}0.4cm,$ 5${\mu}m$, Merck Co.); mobile phase, a mixture of acetonitrile/water/isopropanol (80/5/15) and acetonitrile/water/isopropanol (80/20/15) with gradient system; and the detector, ELSD. On the basis of the optimum conditions a method for the quantitative analysis of ginsenosides were proposed and another cross-examination was carried out for the validation of the selected analytical method conditions. The coefficient of variances (CVs) on the contents of ginsenoside-$Rg_{1}$, -Re and $-Rb_1$ were lower than $3\%$ and the recovery rates of ginsenosides were $89.4\~95.7\%,$ which suggests that the above extraction and separation conditions may be reproducible and reasonable. For the selected HPLC/ELSD conditions, the CVs on the detector responses of ginsenoside-Rg, -Re and $-Rb_1$) were also lower than $3\%$, the regression coefficients for the calibration curves of ginsenosides were higher than 0.99 and two adjacent ginsenoside peaks were well separated, which suggests that the above HPLC/ELSD conditions may be good enough for the determination of ginsenosides.

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Changes of Chemical Characteristics of Soil Solution In Paddy Field from Fifty-Eight Years Fertilization Experiments

  • Kim, Myung Sook;Kim, Yoo Hak;Park, Seong Jin;Lee, Chang Hoon;Yun, Sun Gang;Sonn, Yeon Kyu
    • Korean Journal of Soil Science and Fertilizer
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    • v.48 no.1
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    • pp.22-29
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    • 2015
  • The objectives of this study were to monitor the changes in soil solution nutrients and to evaluate their effect on rice uptake and yield. The changes of chemical characteristics of paddy soil solution were examined from the 58th fertilization experiment in which the continuous rice cropping experiment started in 1954 at the National Academy of Agricultural Science. The treatments were no fertilization (No fert.), inorganic fertilization (NPK), inorganic fertilizer plus rice straw compost (NPKC) and inorganic fertilizer plus silicate and lime fertilizer as a soil amendment (NPKCLS). The fertilizers were added at rates of standard fertilizer application rate in which nitrogen (N), phosphate ($P_2O_5$), potassium ($K_2O$), and sililcate ($SiO_2$) were applied at rates of $75{\sim}150kg\;ha^{-1}$, $70{\sim}86kg\;ha^{-1}$, $75{\sim}86kg\;ha^{-1}$, and $7.5Mg\;ha^{-1}$ respectively and lime was applied to neutralize soil acidity until 6.5. Average Electrical Conductivity (EC) of soil solution in NPKCLS and NPKC ranged from 1.16 to $2.00dS\;m^{-1}$. The $NH{_4}^+$ and $K^+$ levels in NPKCLS and NPKC were higher than that of the other treatments, due to high supply power of rice straw compost. The content of $H_3SiO{_4}^-$ was higher in NPKCLS because of silicate application. The dominant ions in soil solution were $Ca^{2+}$, $Mg^{2+}$ and $Na^+$ among cations and $HCO{_3}^-$, $SO{_4}^{2-}$, and $Cl^-$ among anions in all treatments. The continuous application of inorganic fertilizers plus rice straw compost (NPKC) and silicate fertilizer (NPKCLS) led to the changes of various chemical composition in soil solutions. Also, they had a significant impact on the improvement of rice inorganic uptake and grain yield. Especially, inorganic uptake by rice in NPKC and NPKCLS significantly increased than those in NPK plot; 14~46% for T-N, 32~36% for P, 43~57% for K, and 45~77% for Si. Therefore, the combined application of inorganic fertilizers with organic compost as a soil amendment is considered as the best fertilization practice in the continuous rice cropping for the improvement of crop productivity and soil fertility.

A Multi-Degree of Freedom Measurement System for Determining Geometric Errors in Miniaturized Machine Tool (소형공작기계의 기하학적 오차 평가를 위한 다자유도 측정시스템)

  • S. H., Kweon;Y., Liu;S. H., Yang
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2004.10a
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    • pp.638-643
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    • 2004
  • 소형화된 기계가공시스템은 사용재료의 다양화와 에너지 및 공간의 감소와 같은 장점을 가지고 작고 정밀한 부품을 가공할 수 있는 시스템으로 주목받고 있다. 이러한 시스템이 비록 그 크기가 일반적인 가공시스템에 비해 작지만 정렬 및 조립공정, 기계요소의 불완정성에 의한 기하학적 오차는 여전히 존재한다. 기하학적 오차 평가는 기계시스템의 정밀도를 효과적으로 적은 비용으로 향상시킬 수 있는 오차보정기술을 적용할 수 있는 토대가 된다. 일반적으로, 3 축의 직선축으로 이루어진 공작기계는 21 개의 오차요소를 가진다. 레이져간섭계는 이러한 오차요소를 평가하는데 널리 사용되고 있지만 광학계를 정렬하고 설치하는 데 많은 어려움이 있으며 한번의 설치로 한 개의 오차요소만이 측정 가능하다. 또한, 소형공작기계의 경우, 그 크기로 인해 기존의 레이져 간섭계를 직접적으로 적용할 수 없다. 따라서, 본 연구에서는 소형공작기계를 포함한 소형가공시스템의 기하학적 오차 평가를 위한 새로운 다자유도 측정시스템을 제안하였다. 5 개의 정전용량변위센서를 사용하는 이 시스템을 통해 한 축의 움직임에 따른 5 개의 오차요소를 동시에 측정 가능하다. 균질 변환행렬을 이용한 측정알고리듬을 구성하고 이를 모의시험을 통해 평가하였다. 수학적 모델링을 통해 각 센서의 출력값을 유도하고 이를 이용하여 각 오차요소를 계산하기 위한 식을 유도하였다. 여기서, 단순화된 식을 적용한 경우, 임의의 오차에 대한 측정 알고리듬의 정확도를 평가하였다. 또한, 측정 시스템의 설치시 발생하는 셋업오차에 대한 측정 알고리듬의 민감도 분석을 행하였다. 제안하는 측정 시스템은 구조가 간단하고 고가의 부가장비가 필요치 않다. 또한, 적은 비용으로 구성할 수 있으며 높은 측정 정밀도를 가지고 소형가공시스템에 필요한 오차 평가를 행할 수 있다.가 함유된 계란을 생산하고 섭취하였을 때 특정항체들의 결합을 통해 병원성 미생물의 성장이나 군체를 형성하는 것을 무력화시켜 결과적으로 병원균을 감소시키거나 억제시킨다는 점이다. 오늘날 약물에 내성을 지닌 박테리아의 출현으로 질병감염을 막는데 항생제의 사용효과가 점차 감소하고 있기 때문에 이러한 항생제를 대체할 수 있는 방안으로 계란항체를 이용할 수 있다.한 중공 플랜지 형상의 단조 방법 중 보다 적절한 단조방법인 압조 단조에 있어서 일반적으로 사용되고 있는 SM10C에 대한 유한요소 해석을 수행하였으며, 제품의 형상비에 따라 폴딩 결함의 발생 유무를 검토하고, 폴딩 결함 없이 단조하기 위한 중공 플랜지의 형상한계 비를 제시하였다.도 경미하게 나타났으나, 경련이 나타난 쥐에서는 KA만을 투여한 흰쥐와 구별되지 않았다. 이상의 APT의 항산화 효과는 KA로 인한 뇌세포 변성 개선에 중요한 인자로 작용할 것으로 사료되나, 보다 명확한 APT의 기전을 검색하고 직접 임상에 응응하기 위하여는 보다 다양한 실험 조건이 보완되어야 찰 것으로 생각된다. 항우울약들의 항혈소판작용은 PKC-기질인 41-43 kD와 20 kD의 인산화를 억제함에 기인되는 것으로 사료된다.다. 것으로 사료된다.다.바와 같이 MCl에서 작은 Dv 값을 갖는데, 이것은 CdCl$_{4}$$^{2-}$ 착이온을 형성하거나 ZnCl$_{4}$$^{2-}$ , ZnCl$_{3}$$^{-}$같은 이온과 MgCl$^{+}$, MgCl$_{2}$같은 이온종을 형성하기 때문인것 같다. 한편 어떠한 용리액에서던지 NH$_{4}$$^{+}$의 경

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Studies on the Determination Method of Natural Sweeteners in Foods - Licorice Extract and Erythritol (식품 중 감초추출물 및 에리스리톨 분석법에 관한 연구)

  • Hong Ki-Hyoung;Lee Tal-Soo;Jang Yaung-Mi;Park Sung-Kwan;Park Sung-Kug;Kwon Yong-Kwan;Jang Sun-Yaung;Han Ynun-Jeong;Won Hye-Jin;Hwang Hye-Shin;Kim Byung-Sub;Kim Eun-Jung;Kim Myung-Chul
    • Journal of Food Hygiene and Safety
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    • v.20 no.4
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    • pp.258-266
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    • 2005
  • Licorice Extract and Erythritol, food additives used in korea, are widely used in foods as sweetener. Its application for use in food is regulated by the standard and specification for food additives but official analytical method far determination of these sweetener in food has not been established. Accordingly, we has been carried out to set up analytical method of the glycyrrhizic acid in several foods by the way of thin layer chromatography and high performance liquid chromatography glycyrrhizic acid is qualitative anaylsis technique consists of clean-up with a sep-pak $C_{18}$ cartridge, separation of the sweeteners by Silica gel 60 F254 TLC plate using 1-butanol:4Nammonia solution:ethanol (50:20:10) as mobile solvent. Also, the quantitative analysis for glycyrrhizic acid, was performed using Capcell prk $C_{18}$ column at wavelength 254nm and DW:Acetonitrile (62:38 (pH2.5)) as mobile phase. and we has been carried out to set up analytical method of the erythritol in several foods by the way of high performance liquid chromatography. erythritol is qualitative anaylsis technique consists of clean-up with a DW and hexane. The quantitative analysis for erythritol, was performed using Asahipak NH2P-50 column, Rl and DW:Acetonitrile (25:75) as mobile phase. The glycyrrhizic acid results determined as glycyrrhizic acid in 105 items were as follows; N.D$\∼$48.7ppm for 18 items in soy sauce, N.D$\∼$5.3ppm for 12 items in sauce, N.D$\∼$988.93ppm for 15 items in health food, N.D$\∼$180.7ppm for 26 items in beverages, N.D$\∼$2.6ppm for 8 items in alcoholic beverages repectively and ND for 63 items in the ethers. The erythritol results determined as erythritol in 52 items were as follows; N.D$\∼$155.6ppm for 13 items in gm, N.D$\∼$398.1ppm for 12 items in health foods repectively and ND for 45 items in the others.

Altered Regulation of Renal Acid Base Transporters in Response to Ammonium Chloride Loading in Rats

  • Kim, Eun-Young;Choi, Joon-Seok;Lee, Ko-Eun;Kim, Chang-Seong;Bae, Eun-Hui;Ma, Seong-Kwon;Kim, Suhn-Hee;Lee, Jong-Un;Kim, Soo-Wan
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.2
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    • pp.91-95
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    • 2012
  • The role of the kidney in combating metabolic acidosis has been a subject of considerable interest for many years. The present study was aimed to determine whether there is an altered regulation of renal acid base transporters in acute and chronic acid loading. Male Sprague-Dawley rats were used. Metabolic acidosis was induced by administration of $NH_4Cl$ for 2 days (acute) and for 7days (chronic). The serum and urinary pH and bicarbonate were measured. The protein expression of renal acid base transporters [type 3 $Na^+/H^+$ exchanger (NHE3), type 1 $Na^+/{HCO_3}^-$ cotransporter (NBC1), Na-$K^+$ ATPase, $H^+$-ATPase, anion exchanger-1 (AE-1)] was measured by semiquantitative immunoblotting. Serum bicarbonate and pH were decreased in acute acid loading rats compared with controls. Accordingly, urinary pH decreased. The protein expression of NHE3, $H^+$-ATPase, AE-1 and NBC1 was not changed. In chronic acid loading rats, serum bicarbonate and pH were not changed, while urinary pH was decreased compared with controls. The protein expression of NHE3, $H^+$-ATPase was increased in the renal cortex of chronic acid loading rats. These results suggest that unaltered expression of acid transporters combined with acute acid loading may contribute to the development of acidosis. The subsequent increased expression of NHE3, $H^+$-ATPase in the kidney may play a role in promoting acid excretion in the later stage of acid loading, which counteract the development of metabolic acidosis.

Effect of Different Rumen-degradable Carbohydrates on Rumen Fermentation, Nitrogen Metabolism and Lactation Performance of Holstein Dairy Cows

  • Khezri, A.;Rezayazdi, K.;Mesgaran, M. Danesh;Moradi-Sharbabk, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.5
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    • pp.651-658
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    • 2009
  • Four multiparous lactating Holstein cows fitted with rumen cannulae were fed diets varying in the amount and source of rumen-degradable carbohydrates (starch vs. sucrose) to examine their effects on rumen fermentation, nitrogen metabolism and lactation performance. A $4{\times}4$ Latin square with four diets and four periods of 28 days each was employed. Corn starch and sucrose were added to diets and corn starch was replaced with sucrose at 0 (0 S), 2.5 (2.5 S), 5.0 (5.0 S) 7.5% (7.5 S) of diet dry matter in a total mixed ration (TMR) containing 60% concentrate and 40% forage (DM basis). Replacing corn starch with sucrose did not affect (p>0.05) ruminal pH which averaged 6.41, but the ruminal pH for 7.5 S decreased more rapidly at 2 h after morning feeding compared with other treatments. Sucrose reduced ($p{\leq}0.05$) ruminal $NH_3-N$ concentration (13.90 vs. 17.09 mg/dl) but did not affect peptide-N concentration. There was no dietary effect on total volatile fatty acids (110.53 mmol/L) or the acetate to propionate ratio (2.72). No differences (p>0.05) in molar proportion of most of the individual VFA were found among diets, except for the molar proportion of butyrate that was increased ($p{\leq}0.05$) with the inclusion of sucrose. Total branched chain volatile fatty acids tended to increase ($p{\geq}0.051$) for the control treatment (0 S) compared with the 7.5 S treatment. Dry matter intake, body weight changes and digestibility of DM, OM, CP, NDF and ADF were not affected by treatments. Sucrose inclusion in the total mixed ration did not affect milk yield, but increased milk fat and total solid percentage ($p{\leq}0.05$). Sucrose tended ($p{\geq}0.063$) to increase milk protein percentage (3.28 vs. 3.05) and reduced ($p{\leq}0.05$) milk urea nitrogen concentration (12.75 vs. 15.48 mg/dl), suggesting a more efficient utilization of the rapidly available nitrogen components in the diet and hence improving nitrogen metabolism in the rumen.

Effects of Ammonium, and Nitrate on Callus Growth of Tobacco and Soybean and Activities of Nitrogen Metabolizing Enzymes (암모늄태와 질산태 질소가 담배와 콩의 조직배양시 생육 및 질소대사 효소의 활성에 미치는 영향)

  • 박혜선;설종호;장매희
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.57-61
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    • 1998
  • Soybean and tobacco tissue cultured with modified MS media containing 4 different ratio(as N) of nitrate to ammonium combination which were 3:0, 2:1, 1:2 and 0:3. The highest callus growth in soybean were observed in the 2:1 medium. The medium containing nitrate only was detrimental to soybean callus growth. Tobacco callus grown with nitrate-only grew as well as those in the 2:1 and very slowly with ammonium-only. In tobacco callus, the total nitrogen in the callus increased with the increase of nitrogen concentration in the medium, but in soybean callus, the opposite result was noted. Nitrate reductase activity in tobacco callus was high when grown with nitrate-only but low with ammonium-only. In case of soybean callus, nitrate reductase activity was high in the 2:1 and remarkably low in nitrate-only medium. Both in soybean and tobacco callus, the activity of glutamine synthetase was high with nitrate-only, but low with ammonium.

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Trophic State and Water Quality in Major Lakes of the Sumjin and Youngsan River Systems (섬진강 ${\cdot}$ 영산강 수계 주요 호소의 수질 동향과 영양상태 조사)

  • Yi, Sang-Hyon;Chang, Nam-Ik;Kim, Jong-Min;Kim, Hyun-Ku;Cho, Young-Gwan;Jeong, Jin;Sin, Yong-Sik
    • Korean Journal of Ecology and Environment
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    • v.39 no.3 s.117
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    • pp.296-309
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    • 2006
  • This study aimed to analyze water quality (temperature, secchi depth, DO, $NH_4$, $NO_3$, $PO_4$, TN, TP, chlorophyll a) and trophic state index during the period of year 2000 ${\sim}$ 2004 in Lake Juam, Lake Dongbok and Lake Youngsan. Lakes Juam and Dongbok except Lake Youngsan were stratified during warm seasons. Water turbidity estimated by secchi disk depth was the highest in Lake Youngsan compared with other lakes. DO concentrations were low in the bottom water when chlorophyll a was high in Lake Juam and Dongbok. Nutrient concentrations were higher in Lake Youngsan than other lakes whereas chlorophyll a was highest in Lake Dongbok. Lake Youngsan was the most eutrophic compared to other two lakes based on the Trophic State Idex (TP) and TSI (SD), The TSI (CHL) was high but the TSI (TP) were low in Lake Juam and Dongbok. These results suggest phytoplankton may be limited by phosphates (P) in Lake Juam and Dongbok whereas light availability in the water column may affect growth of phytoplankton in Lake Youngsan.

Study on Soil Extraction Methods for the Human Health Risk Assessment of Crop Intake Pathway around Abandoned Metal Mine Areas (폐금속광산 지역 농작물섭취경로의 인체위해도 산정을 위한 생물농축계수와 토양분석방법에 관한 연구)

  • Lim, Tae-Yong;Lee, Sang-Woo;Yun, Seong-Taek;Kim, Soon-Oh
    • Journal of Soil and Groundwater Environment
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    • v.21 no.6
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    • pp.56-66
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    • 2016
  • Generally, the contribution of crop-intake pathway (CIP) is remarkable in human health assessment (HHA) of heavy metal contamination. Although the crop exposure concentrations (Cp) should directly be used for calculating the average daily dose (ADD) of CIP, the soil exposure concentration (Cs) multiplied by soil-crop bio-concentration factor (BCF) has frequently been used instead of using Cp values. Thus, the BCF values are significant in the HHA, and care should be taken to ensure the reasonable acquisition of BCF values. Meanwhile, the BCF values are known to be significantly affected by analytical methods. Nevertheless, they have been calculated from the concentrations of soil and crop analyzed by only one method: total digestion (aqua regia extraction). For this reason, this study was initiated to seek appropriate soil analysis methods for effective computation of the ADD of CIP. The concentrations of 5 metal contaminants (As, Cd, Cu, Pb, and Zn) in 127 soil samples obtained from 4 abandoned metal mine areas were analyzed by several methods including total digestion and partial digestions using 0.1/1 N HCl, 1M $NH_4NO_3$, 0.1 M $NaNO_3$, and 0.01M $CaCl_2$. The heavy metal concentrations in 127 crop samples (rice grains) were analyzed by total digestion as well. Using the concentrations of soils and crops, the BCF values of each contaminant were calculated according to the kind of soil extraction methods applied. Finally, the errors between Cp and $C_s{\times}BCF$ were computed to evaluate the relevance of each method. The results indicate that the partial extraction using 0.1 N and 1 N HCl was superior or equivalent to total digestion. In addition, the 0.1M $NaNO_3$ method combined with total digestion is recommended for improving the reliability of BCF values.