초록 ▼

$\bullet$ 과학적인 피해조사 및 평가시스템 구축
- 사고유류 확인을 위한 첨단 핵심분석기술 개발의 일환으로 소형선박 연료유 오염지표 화합물인 세스키터페인(sesquiterpane) 분석기술 개발.
- 유류오염에 의한 생물 영향평가 기술개발의 일환으로 유처리제로 분산된 유류에 노출된 수산물의 영향여부 입증 기술 확립.
$\bullet$ 유류오염 다발해역에 대한 유류오염 자료 확보
- 부산 해역의 유류계 PAHs 오염실태 파악 및 D/B 구축을 위해 표층퇴적물 및 어패

$\bullet$ 과학적인 피해조사 및 평가시스템 구축
- 사고유류 확인을 위한 첨단 핵심분석기술 개발의 일환으로 소형선박 연료유 오염지표 화합물인 세스키터페인(sesquiterpane) 분석기술 개발.
- 유류오염에 의한 생물 영향평가 기술개발의 일환으로 유처리제로 분산된 유류에 노출된 수산물의 영향여부 입증 기술 확립.
$\bullet$ 유류오염 다발해역에 대한 유류오염 자료 확보
- 부산 해역의 유류계 PAHs 오염실태 파악 및 D/B 구축을 위해 표층퇴적물 및 어패류를 각각 40개, 20개 정점에서 채취하여 유류계 PAHs를 분석.
- 분석방법은 해양환경공정시험방법에 준하여 실시하며, 정도관리 프로그램에 의거하여 분석.

Abstract ▼

1. Analytical method setup and environmental occurrence of sesquiterpanes
$\bullet$ Sesquiterpanes ranges from C14 to C16 were analysed using gas chromatography-mass spectrometry. Detection limit for sesquiterpanes ranged from 0.2 ng/g to 0.5 ng/g depending on the compounds.
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1. Analytical method setup and environmental occurrence of sesquiterpanes
$\bullet$ Sesquiterpanes ranges from C14 to C16 were analysed using gas chromatography-mass spectrometry. Detection limit for sesquiterpanes ranged from 0.2 ng/g to 0.5 ng/g depending on the compounds.
$\bullet$ For the identification of sesquiterpanes, retention characteristics, target ions and spectrums from previous works were used. Calibration curve was made using decalin and $decalin-_{d18}$, and target sesquiterpanes were quantitated by relative response factor between each compound and $decalin-_{d18}$.
$\bullet$ Total sesquiterpanes in the 40 kinds of imported crude oils had the ranges of $0.29{\sim}8.38\;mg/g$, whose concentration and indices varied depending on their origin.
$\bullet$ Sesquiterpane concentrations in 14 marine fuel oil ranged from 0.27 mg/g to 4.02 mg/g, whose concentrations and indices varied depending on their raw material`s origin and refineries.
$\bullet$ Sesquiterpanes in sediment samples from small port varied 129 ng/g to 818 ng/g, whose indices were relatively similar indicating the similar input source.
$\bullet$ Forty sediment samples from Pusan area were analyzed for sesquiterpanes. Their concentration had ranges of not detected ${\sim}429\;ng/g$ and showed significant correlation with total PAHs, indicating the influence of the petrogenic PAHs.
2. Effects of dispersed crude oil to fish: biomarker approach
$\bullet$ The use of oil dispersant is a controversial countermeasure in the effort to minimize the impact of oil spills. To evaluate the relative effects of fish exposed to water-accommodated fraction of oil, WAFs and chemically enhanced WAF(dispersant: Corexit 9500 and one domestic product), CYP1A induction in rockfish were measured.
$\bullet$ To evaluate expression of hepatic detoxification enzyme, the cDNA library were constructed after the b-NF exposure to rockfish. CYP1A primers designed to a conserved region in GenBank were used. The deduced amino acid sequences of the CYP1A were submitted to Genbank(Genbank no. AY568556).
$\bullet$ Level of CYP1A and EROD activity in rockfish exposed to CEWAF(Corexit 9500) increased in response to concentration of 0.1 %(v/v) after 48 h. After 72 h of exposure, level of CYP1A and EROD activity increased at all concentration level of CEWAF(Corexit 9500 and A domestic product) significantly.
$\bullet$ The uptake of polycyclic aromatic hydrocarbones(PAHs) by fish can be determined by screening the PAH metabolites in fish file. In the study, a bile sample exposed CEWAF showed the 350 nm peak, which is attributable to 1-OH-py. But sample exposed to WAF showed low fluorescence at 350 nm. Fish samples exposed to both Corexit 9500 and A product CEWAF showed highly increased fluorescence responses than fish exposed WAF and control fish after 72 h of exposure.
$\bullet$ These experiments suggest that the use of oil dispersants will increase the harmful effects of spilled oil to fish, which can be effectively detected using CYP1A enzyme system assay.
3. Monitoring of petrogenic PAHs in coastal regions of Busan and nearby areas
$\bullet$ The background distribution of PAHs in the surface sediment and bivalves samples from Busan area had been investigated.
$\bullet$ Total PAHs concentration in surface sediments were in the range of $5.72{\sim}953\;ng{\cdot}g^{-1}$ (mean value; $175\;ng{\cdot}g^{-1})$. Levels of PAHs in this study is similar to those from national monitoring study.
$\bullet$ Total PAHs concentration in bivalves were in the range of $71.8{\sim}742\;ng{\cdot}g^{-1}$ (mean value; $232\;ng{\cdot}g^{-1})$. Lipid normalized PAHs concentration were in the range of $1{\sim}10\;ug{\cdot}g^{-1}$.
$\bullet$ Elevated level of PAHs were found at harbor and nearby regions where various ship related activities exist. And source recognition index (FLRT/PYR vs PHEN/ANTH) indicated that burning of fossil fuel is the major source of PAHs in this region. But the signal of petrogenic sources was also found at the harbor.
$\bullet$ This study will contribute to the source identification and risk assessment when unexpected oil spill accident occurs in this region, where the biggest harbor in Korea, Busan harbor is located.

목차 Contents

• 제 1 장 연구개발과제의 개요...24
• 제 1 절 연구개발의 목적...26
• 제 2 절 연구개발의 필요성...26
• 제 3 절 연구개발의 내용 및 범위...30
• 제 2 장 국내외 기술개발현황 및 과학기술정보...32
• 제 1 절 국내외 기술개발현황...34
• 제 2 절 현 기술상태의 취약성...37
• 제 3 절 앞으로의 전망...38
• 제 3 장 연구개발 수행내용 및 결과...40
• 제 1 절 세스키터페인 분석기술 개발 및 적용...42
• 1. 서론...42
• 2. 재료 및 시약...44
• 3. 분석방법의 정립...44
• 가. 유류 시료...44
• 나. 퇴적물 시료...50
• 4. 유류시료내 세스키터페인의 분포특성...52
• 가. 국내 수입원유...53
• 나. 해상용 연료유...56
• 다. 세스키터페인을 활용한 지수노출과정...58
• 5. 환경시료내 세스키터페인의 분포특성...63
• 가. 소형 어항...63
• 나. 부산해역...65
• 제 2 절 유처리제로 분산된 유류에 노출된 유용수산물의 생물영향 평가...71
• 1. 서론...71
• 2. 재료 및 방법...77
• 가. 유류와 유분산제...77
• 나. 실험어...79
• 다. 노출실험...79
• 라. 노출과정...82
• 마. CYP1A 유전자 발현 조사를 위한 cDNA 라이브러리 제작...82
• 바. 형광스펙트럼을 이용한 1-hydroxypyrene분석...83
• 사. Total mRNA추출과 $poly(A)^+$ mRNA 정제...83
• 아. DNA sequencing과 분석...87
• 자. Northern blotting 분석...87
• 차. EROD(ethoxyresorufin O-deethylase)의 활성측정...90
• 카. CYP1A 단백질 발현의 측정...90
• 타. 뇌와 근육의 AChE 활성 분석...90
• 파. 단백질 함량의 정량...91
• 3. 결과 및 고찰...93
• 4. 결론...97
• 제 3 절 부산해역 내 다환방향족탄화수소의 분포특성...116
• 1. 조사지역개관...116
• 가. 부산 해역의 특성...116
• 나. 해안 방제 자료에 따른 지역 정보...119
• 2. 시료채취 및 분석방법...138
• 가. 시료채취...138
• 나. PAHs 분석...144
• 다. 자료질검정(quality assurance/quality control; QA/QC)...152
• 3. 분석결과 및 고찰...154
• 가. 표층퇴적물 내 PAHs...154
• 나. 이매패류 내 PAHs...166
• 제 4 장 연구개발 목표달성도 및 관련분야에의 기여도...178
• 제 1 절 연구개발 목표달성도...180
• 제 5 장 연구개발 결과의 활용계획...182
• 제 6 장 참고문헌...186
• 부록...194