표제지
목차
List of abbreviation 10
I. 서론 11
II. 실험재료 및 방법 17
1. 시약 및 기기 17
2. 표준액 조제 20
3. 뇨 중 아크릴아마이드 및 대사체 분석법 개발 20
3.1. 시료의 전처리 20
3.2. LC-ESI-MS/MS 분석 조건 22
3.3. 분석법 검증(Validation) 23
4. 인체모니터링에 응용 26
4.1. 소변 시료의 채집 및 처리 26
4.2. 모니터링 수행 시 정량 및 평가 26
4.3. 시료군(batch)의 정도관리 26
III. 결과 및 고찰 29
1. 뇨 중 아크릴아마이드 및 대사체 분석법 개발 29
1.1. 시료의 전처리 29
1.2. LC-ESI-MS/MS 분석법 확립 37
1.3. 아크릴아마이드의 정성적 확인 46
1.4. 바탕시료에서 아크릴아마이드 검출 48
2. 분석법 검증(Validation) 53
2.1. 특이성 53
2.2. 직선성, 검출한계 및 정량한계 54
2.3. 정확성 및 정밀성 56
2.4. 회수율 57
3. 인체모니터링 결과 및 고찰 58
3.1. 인체모니터링 결과 58
3.2. 외국의 모니터링 결과와 비교 및 고찰 60
3.3. 지역별, 연령별 그리고 성별 비교 62
IV. 결론 67
V. 참고문헌 69
VI. 국문초록 73
VII. Abstract 75
Table 1. Analytical methods review for determination of acrylamide in food 14
Table 2. Analytical methods review for determination of acrylamide and metabolites in urine 15
Table 3. Evaluation of volatility before and after drying under a nitrogen stream (peak area) 30
Table 4. Comparison of different types of SPE cartridge (peak area) 32
Table 5. Comparison of elution solvent volume (peak area) 35
Table 6. Comparison of injection volume (peak area) 43
Table 7. LC-ESI-MS/MS conditions for acrylamide and its metabolites 45
Table 8. Result of blank test to evaluate reproducibility 52
Table 9. Calibration curves, linearity, LOD, and LOQ 55
Table 10. Intra- and inter-day precision and accuracy of AA, GA and AAMA 56
Table 11. Recovery of analytes spiked in artificial urine 57
Table 12. Concentration of AA, GA and AAMA in korea population 59
Table 13. Comparison concentration of acrylamide and metabolites measured korea population (n=1903) with monitoring study results in other paper 61
Table 14. Result of comparison mean concentration of acrylamide and metabolites measured Korean population (n=1903) according to region 63
Table 15. Result of comparison concentration of acrylamide and metabolites measured Korean population (n=1903) according to ages 65
Table 16. Result of comparison concentration of acrylamide and metabolites measured Korean population (n=1903) according to sex 66
Fig. 1. Possible routes to acrylamide formation in foodstuff. 12
Fig. 2. Metabolic pathway of acrylamide with respect to the formation of mercapturic acids. 13
Fig. 3. Structure of acrylamide and its metabolites 19
Fig. 4. Analysis order for quality control in one batch 28
Fig. 5. Evaluation of volatility before and after drying under a nitrogen stream 30
Fig. 6. Comparison of different types of SPE cartridge 32
Fig. 7. Selection of buffer pH for the sample loading step 33
Fig. 8. Selection of solvent for the washing step 34
Fig. 9. Scheme of sample preparation 36
Fig. 10. Precursor ion and product ion scan spectra of (A) AA (B) GA produced by LC-ESI-MS/MS 38
Fig. 11. Effect of buffer concentration in the mobile phase (retention time) 41
Fig. 12. Effect of buffer concentration in the mobile phase (peak area) 41
Fig. 13. Comparison of mobile phase additive 42
Fig. 14. MRM chromatogram of AA and AA-d₃ (I.S.) in real urine sample at a flow rate of (A) 150 μL/min and (B) 100 μL/min 44
Fig. 15. MRM chromatogram for confirmation of identity of AA (A) before spiking AA standard and (B) after spiking AA standard in real urine 47
Fig. 16. Result of blank test of (a) HLB type, (b) MCX type and (c) ENV+ type 49
Fig. 17. MRM chromatograms of analytes (A) spiked in artificial urine and (B) in real urine sample (FG-03-18) 53
Fig. 18. Calibration curves of AA, GA and AAMA generated from spiked artificial urine sample 55
Fig. 19. The detection frequency of acrylamide and metabolites measured in korean population 59