대한환경공학회 2010 춘계학술연구발표회 영산강자연유기물질발생기원규명을 위한특이 Molecules 측정 전강민 황동진 * 조재원 광주과학기술원환경공학과대안수처리엔지니어링연구실, * 영산강물환경연구소
연구배경및목적 연구배경 연구목적
연구배경 자연유기물질수계유입경로 비 농경지 산림지역 도시지역 하수처리장 미생물및생물활동부산물및사체 토양 (http://www.ozcoasts.org.au) 3/24
연구배경 자연유기물질특성 지역별, 계절별, 수계별, 국가별로다른특성 발생기원, 계절영향, 화학적, 생물학적변화반응영향에따라다른특성 => 자연유기물질의분자구조, 반응및거동특성변화 자연유기물질의역활 탄소와질소, 인, 황과같이생물체를구성하는주요성분들의생태계순환에관여 수계에존재하는다른유기및무기화합물과리간드를형성하여오염물질의독성을증감 4/24
연구목적 영산강수계에존재하는자연유기물질의특이 Molecules 을선정하고, 이를화학적으로정밀분석 영산강자연유기물질의발생기원을규명하고, 이를토대로하여영산강수계고유의자연유기물질에대한기초자료를제공하고자함 5/24
시료채취및분석방법 시료채취 분석방법
시료채취 11 개주요지점선정 1. 담양 (DY) 2. 오치 (WC) 3. 광주 1 (GJ1) 4. 광주천 2 (GJS2) 5. 하수처리장방류수 (WE) 6. 광주 2-1 (GJ2) 7. 활룡강 (HRS) 8. 지석천 (JSS) 9. 광산 (GS) 10. 나주 (NJ) 11. 무안 (MA) 7/24
분석방법 Categories Methods Detailed information DOC TOC measurement 800 TOC analyzer, Sievers UV absorbance UVA at 254 nm UV-1601, Shimadzu TN TN measurement TOC-V CPH equipped with TNM-1, Shimadzu Anion and Cation IC ICS-90/DX-120, Dionex Metal and Metalloids ICP-MS 7500ce, Agilent Column: Protein pak 125, Waters MW distribution Component type and complexity HP-SEC 3D FEEM UV/Vis detector: SPD-10A VP, Shimadzu Fluorescence detector: RF-10A XL, Shimadzu Fluorescence spectrophotometer, F-2500, Hitachi NOM structure NOM fractionation XAD 8/4 resins, Amberlite HPLC: LC-9201, JAI Recycling preperative HPLC NOM separation RI detector: RI-50, JAI UV detector: JAI UV 3072, JAI Column: GEL-GS 310, JAI 8/24
실험결과및고찰 Water characteristics High performance size-exclusion chromatography 3D Fluorescence Excitation-Emission matrix NOM fractionation using XAD 8/4 resins Recycling preperative HPLC
Water characteristics ph Conductivity (µs/cm) DOC (mgc/l) UV 254 (cm -1 ) SUVA (L mg -1 m -1 ) 담양 7.3 416.3 2.66 (±0.04) 0.0348 (±0.0006) 1.31 (±0.00) 우치 7.7 370.9 3.30 (±0.02) 0.0475 (±0.0007) 1.44 (±0.03) 광주 1 8.1 356.8 3.82 (±0.02) 0.0494 (±0.0000) 1.29 (±0.01) 광주천 2 8.1 452.3 6.11 (±0.01) 0.0709 (±0.0001) 1.16 (±0.00) 하수처리장방류수 6.8 564.5 7.35 (±0.03) 0.1003 (±0.0000) 1.36 (±0.01) 광주 2-1 7.4 522.8 7.93 (±0.01) 0.0869 (±0.0001) 1.10 (±0.00) 황룡강 8.7 271.1 2.93 (±0.03) 0.0417 (±0.0011) 1.42 (±0.02) 지석천 8.3 353.4 3.71 (±0.02) 0.0456 (±0.0001) 1.23 (±0.01) 광산 7.7 474.1 6.19 (±0.0.1) 0.0718 (±0.0001) 1.16 (±0.00) 나주 8.1 448.7 5.83 (±0.04) 0.0695 (±0.0003) 1.19 (±0.00) 무안 8.0 1,325.0 4.10 (±0.02) 0.0685 (±0.0012) 1.67 (±0.04) 10/24
Nitrogen characteristics TN (mg/l) NO 2 - (mgn/l) NO 3 - (mgn/l) NH 4 + (mgn/l) org-n (mgn/l) org-n/c (M) 담양 3.57 (±0.09) 0.00 0.76 0.00 2.81 0.91 우치 2.96 (±0.00) 0.00 0.58 0.05 2.32 0.60 광주 1 3.05 (±0.06) 0.00 0.53 0.19 2.33 0.52 광주천 2 7.15 (±0.23) 0.15 1.12 1.36 4.53 0.64 하수처리장방류수 11.28 (±0.28) 0.05 1.35 3.44 6.44 0.75 광주 2-1 10.11 (±0.05) 0.13 0.77 4.69 4.52 0.49 황룡강 2.15 (±0.01) 0.00 0.40 0.00 1.75 0.51 지석천 5.02 (±0.02) 0.00 1.36 0.47 3.19 0.74 광산 9.63 (±0.13) 0.10 0.80 3.72 5.01 0.69 나주 9.22 (±0.10) 0.07 0.90 3.30 4.94 0.73 무안 3.13 (±0.05) 0.00 0.59 0.00 2.53 0.53 11/24
High performance size-exclusion chromatography MW distribution of aromatic substances 25000 20000 15000 10000 담양우치광주 1 광주천 2 하수처리장방류수광주 2-1 황룡강지석천광산나주무안 2 620 750 1010 1100 1370 1440 UV response (mv) 5000 0 1e+1 1e+2 1e+3 1e+4 1e+5 Molecular weight (Da) 좁은범위의분자량크기분포를나타냄 12/24
High performance size-exclusion chromatography MW distribution of protein-like substances 80000 60000 40000 담양우치광주 1 광주천 2 하수처리장방류수광주 2-1 황룡강지석천광산나주무안 2 230 330 610 57710 20000 Fluorescence resp 0 1e+0 1e+1 1e+2 1e+3 1e+4 1e+5 Molecular weight (Da) 방향성물질보다작은분자량크기분포를나타냄 13/24
3D fluorescence excitation-emission matrix Fluorescence contour plots of standard substances: (a) BSA, (b) SRHA, and (c) SRFA The maximum peak of SRHA at Ex=270 nm/em=450 nm and Ex=320 nm/em=440 nm The maximum peak of BSA at Ex=280 nm/em=340 nm 500 The maximum peak of SRFA at Ex=260 nm/em=440 nm and Ex=320 nm/em=440 nm 500 (a) 500 (b) (c) 450 450 450 400 400 400 350 350 350 250 250 300 350 400 Emission (nm) 450 500 Excitation (nm) Excitation (nm) Excitation (nm) 300 300 250 250 300 350 400 450 500 300 250 250 Emission (nm) Bovine serum albumin (BSA) (Sigma-aldrich, USA) Suwannee River humic acid (SRHA) (International Humic Substances Society, USA) Suwannee River fulvic acid (SRHA) (International Humic Substances Society, USA) 14/24 300 350 400 Emission (nm) 450 500
3D fluorescence excitation-emission matrix Ex (nm) Em (nm) Type of compounds 담양 330 410 Humic-like substances 우치 315 405 Humic-like substances 광주1 290 400 Humic-like substances 광주천2 280 340 Protein-like substances 310 410 Humic-like substances 하수처리장방류수 280 340 Protein-like substances 320 400 Humic-like substances 광주2-1 280 340 Protein-like substances 320 400 Humic-like substances 황룡강 310 400 Humic-like substances 지석천 310 400 Humic-like substances 광산 280 340 Protein-like substances 310 410 Humic-like substances 나주 280 340 Protein-like substances 315 400 Humic-like substances 무안 310 400 Humic-like substances 15/24
NOM fractionations using XAD 8/4 resins 60 57 Hydrophobic Hydrophilic Transphilic Fractions (%) 50 40 30 20 29 14 42 46 47 36 35 23 18 34 20 10 0 담양하수처리장방류수광주 2-1 무안 영산강수계의자연유기물질형성에 allochthonous 유기물보다 autochthonous 유기물보다는이더큰영향을주었음 16/24
Recycling preperative HPLC 특이 Molecules 의분리 : RI : UV III : RI : UV 담양 I II IV 광주 2-1 Operating conditions Mobile phase DI Water Pressure 14 kgf/cm 2 : RI : UV 하수방류수 : RI : UV 무안 Injection volume Flow rate 3 ml 5 ml/min 17/24
Recycling preperative HPLC 특이 Molecules 의분리 RI III I II IV UV II III IV I 18/24
Recycling preperative HPLC DOC (mgc/l) TN (mg/l) UV 254 (cm -1 ) SUVA (L mg -1 m -1 ) Fe (µg/l) 담양 1 1.76 (±0.07) 0.28 (±0.00) 0.0344 (±0.0001) 1.96 (±0.08) 10.9 (±0.5) 담양 2 4.95 (±0.11) 0.77 (±0.01) 0.0993 (±0.0001) 2.01 (±0.04) 14.1 (±1.0) 담양 3 3.36 (±0.10) 5.64 (±0.03) 0.0647 (±0.0000) 1.93 (±0.06) 20.1 (±0.5) 담양 4 9.77 (±1.88) 69.24 (±0.10) 0.2050 (±0.0000) 2.14 (±0.41) 10.4 (±0.1) 담양원수 5.20 (±0.00) 9.48 (±0.03) 0.0896 (±0.0001) 1.72 (±0.00) 5.5 (±0.7) 하수방류수 1 3.91 (±0.08) 0.44 (±0.01) 0.0637 (±0.0000) 1.63 (±0.03) 19.9 (±0.2) 하루방류수 2 9.11 (±0.11) 10.92 (±0.01) 0.1592 (±0.0001) 1.75 (±0.02) 30.4 (±0.2) 하루방류수 3 7.43 (±0.10) 21.15 (±0.00) 0.1461 (±0.0000) 1.97 (±0.03) 25.0 (±0.2) 하루방류수 4 12.44 (±0.05) 60.19 (±0.25) 0.3382 (±0.0003) 2.72 (±0.01) 19.1 (±1.8) 하루방류수원수 7.34 (±0.01) 8.95 (±0.03) 0.1329 (±0.0001) 1.81 (±0.00) 8.8 (±3.6) 광주 2-1 1 3.63 (±0.02) 0.40 (±0.01) 0.0565 (±0.0000) 1.56 (±0.01) 7.6 (±1.3) 광주 2-1 2 3.99 (±0.12) 0.62 (±0.01) 0.0680 (±0.0001) 1.70 (±0.05) 58.9 (±12.4) 광주 2-1 3 2.92 (±0.11) 1.65 (±0.02) 0.0523 (±0.0001) 1.79 (±0.06) 10.5 (±0.8) 광주 2-1 4 7.63 (±0.03) 30.75 (±0.18) 0.1807 (±0.0000) 2.37 (±0.01) 9.5 (±0.0) 광주 2-1 4-2 3.01 (±0.13) 0.62 (±0.02) 0.0575 (±0.0000) 1.91 (±0.08) 8.6 (±0.4) 광주 2-1 원수 4.74 (±0.07) 3.98 (±0.01) 0.0819 (±0.0000) 1.73 (±0.02) 5.8 (±1.7) 무안 1 2.51 (±0.02) 0.22 (±0.01) 0.0526 (±0.0001) 2.10 (±0.02) 5.2 (±0.6) 무안 2 5.43 (±0.07) 0.62 (±0.00) 0.1154 (±0.0000) 2.13 (±0.03) 5.3 (±0.1) 무안 3 3.44 (±0.07) 1.44 0.0719 (±0.0001) 2.09 (±0.04) 34.9 (±9.7) 무안 4 4.39 (±0.01) 13.76 0.1112 (±0.0000) 2.53 (±0.01) 13.4 (±0.5) 무안원수 5.09 (±0.02) 4.53 0.1020 (±0.0001) 2.00 (±0.00) 7.0 (±0.6) 19/24
Recycling preperative HPLC 담양 하수방류수 60000 50000 40000 담양 1 담양 2 담양 3 담양 4 담양원수 850 920 1320 80000 60000 하수방류수 1 하수방류수 2 하수방류수 3 하수방류수 4 하수방류수원수 840 910 1340 30000 1820 40000 1830 UV response (mv) 20000 10000 0 1 10 100 1000 10000 Molecular weight (Da) UV response (mv) 20000 0 1 10 100 1000 10000 Molecular weight (Da) Peak 별로분자량크기분포에서차이를나타냄. 20/24
Recycling preperative HPLC 광주 2-1 무안 50000 40000 30000 광주 2-1 1 광주 2-1 2 광주 2-1 3 광주 2-1 4 광주 2-1 4-2 광주 2-1 원수 820 860 900 1380 60000 50000 40000 무안 1 무안 2 무안 3 무안 4 무안원수 770 840 1240 20000 1890 30000 20000 1720 UV response (mv) 10000 0 1 10 100 1000 10000 Molecular weight (Da) UV response (mv) 10000 0 1 10 100 1000 10000 Molecular weight (Da) Peak 별로분자량크기분포에서차이를나타냄. 21/24
요약및향후연구방향 요약 현재연구방향
요약및향후연구방향 요약 항목 DOC 및질소화합물 MW distribution 3DFEEM 주요결과 하수방류수와광주 2-1 지점에서높게나타남 Protein-like 물질들이방향성물질보다작은분자량물질로구성 Humic-like substances 친수성 / 소수성비율 특이 Molecules 특성파악 외국의 NOM 에비해높은친수성비율을나타냄 Peak 별로각기다른 DOC, TN, 및 Fe 농도 => 각각의 peak 형성에서로다른조건들이작용 향후연구방향 FTIR 및 Pyrolysis GC/MS 를이용한분리된특이 Molecules 의기능족구성및구조를파악 23/24
감사의글 분연구는한국과학재단의국가지정연구실사업 (NOM lab: R0A-2007-000-20055-0) 의지원을받아영산강물환경연구소와공동으로수행된연구입니다 Thank you for your attention! 발표자 : 전강민, imonmyway@gist.ac.kr 지도교수 : 조재원, jwcho@gist.ac.kr http://www.gistwetland.org 24/24