목차

국문 초록
Chapter 1 : Literature Review 1
1.1. Polycyclic aromatic hydrocarbons 2
1.1.1. Characteristics of polycyclic aromatic hydrocarbons 2
1.1.2. Benzo[a]anthracene 5
1.1.3. Chrysene 7
1.1.4. Benzo[b]fluroanthene 9
1.1.5. Benzo[a]pyrene 11
1.2. Chemistry and formation of polycyclic aromatic hydrocarbons 13
1.3. Polycyclic aromatic hydrocarbons in herbal medicines 15
1.4. Analysis method using chromatography 17
1.5. Analysis method using enzyme-linked immunosorbent assay 20
1.6. Reduction of the benzo[a]pyrene-induced hepatotoxicity 25
1.7. Conclusions 29
Chapter 2 : Evaluation of polycyclic aromatic hydrocarbons content of herbal medicine products in Korea by HPLC-FLD 30
2.1. Introduction 31
2.2. Materials and methods 33
2.2.1. Chemicals and reagents 33
2.2.2. Sample preparation for analysis of PAH4 33
2.2.3. Clean-up and extraction for herbal medicine products preprocessing 34
2.2.4. HPLC-FLD analysis of PAH4 35
2.2.5. Quantification and identification of PAH4 36
2.2.6. Method validation for analytical quality assurance 36
2.3. Results and discussions 37
2.3.1. Validation of analytical method for PAHs analysis 37
2.3.2. Comparison of PAH4 contents in herbal medicine product samples 45
2.4. Conclusions 53
Chapter 3 : Sandwich enzyme-linked immunosorbent assay (ELISA) to quantify monoclonal antibody (B[a]P-13) for herbal medicine products 54
3.1. Introduction 55
3.2. Materials and methods 58
3.2.1. Chemicals and reagents 58
3.2.2. Preparation of B[a]P derivatives 59
3.2.3. Sampling and extraction procedure for herbal medicines 59
3.2.4. Sandwich ELISA procedure 61
3.2.5. Evaluation of cross-reactivity for determination 63
3.2.6. Matrix effect of the organic solvents 63
3.2.7. Correlation of benzo[a]pyrene detection ELISA compared to HPLC-FLD in herbal medicine products 63
3.2.8. Statistical analysis 64
3.3. Results and discussions 65
3.3.1. Limit of detection, limit of quantification in the ELISA calibration curve 65
3.3.2. Evaluation of cross-reactivity for determination 69
3.3.3. Matrix effect of the organic solvents 72
3.3.4. Correlation of benzo[a]pyrene detection ELISA compared to HPLC-FLD in herbal medicine products 75
3.4. Conclusions 83
Chapter 4 : Reduction of the benzo[a]pyrene-induced hepatotoxicity by neferine in lotus and daidzein, genistein in soybean 84
4.1. Introduction 85
4.2. Materials and methods 87
4.2.1. Chemicals and reagents 87
4.2.2. Sample preparations for extraction of neferine 88
4.2.3. Sample preparations for extraction of daidzein, genistein 89
4.2.4. HepG2 cells culture and treatment 89
4.2.5. Cell viability assay and proliferation assay 90
4.2.6. Cell extraction 90
4.2.7. HPLC-UV analysis for method validation 91
4.2.8. BPDE-DNA adduct formation analysis 92
4.2.9. The typical intracellular metabolites of B[a]P were measured by high performance liquid chromatography (HPLC) 93
4.2.10. Statistical analysis 93
4.3. Results and discussions 94
4.3.1. Reduction of B[a]P-induced hepatotoxicity by neferine, daidzein, genistein 94
4.3.2. Reduction of intracellular B[a]P metabolites by neferine, daidzein, genistein 101
4.3.3. Validation of analytical method for neferine, daidzein, genistein analysis 107
4.3.4. Comparison of neferine, daidzein and genistein concentrations in lotus and soybean samples 115
4.4. Conclusions 122
Chapter 5. Conclusions 123
References 126
Abstract 156