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목차
I. 서론 11
II. 재료 및 방법 13
1. 실험 재료 13
1) 시료 13
2) 정제된 실험재료 15
3) 시약 및 기기 16
2. 실험 방법 19
1) 항산화 효과 측정 19
2) 미백 효과 측정 22
3) 주름개선 효과 측정 23
4) 항염증 효과 측정 25
5) 항균 효과 측정 30
6) W/O/W multiple emulsion 제조 32
7) EGCG를 첨가한 W/O/W multiple emulsion의 안정성 측정 35
8) EGCG를 첨가한 W/O/W multiple emulsion의 유효성측정 39
9) EGCG를 첨가한 W/O/W multiple emulsion의 안전성 측정 40
III. 결과 및 고찰 41
1. 항산화능 측정결과 41
1) 전자공여능 확인 41
2) Superoxide dismutase(SOD) 유사활성 확인 43
3) Superoxide anion radical 소거 능 확인 45
2. 미백 효과 측정 결과 47
1) Tyrosinase 저해 활성 확인 47
3. 주름개선 효과 측정 결과 49
1) Elastase 저해활성 확인 49
2) Collagenase 저해활성 확인 51
4. 항염증 효과 측정 결과 53
1) Hyaluronidase 저해활성 확인 53
2) Nitric oxide 저해활성 확인 55
3) iNOS 단백질의 발현 확인 57
5. 항균 효과 측정 결과 59
1) 생육 저해환(clear zone) 확인 59
6. EGCG를 첨가한 화장품의 안정성 측정결과 62
1) EGCG를 첨가한 W/O/W multiple emulsion 입자 안정성 확인 62
7. EGCG를 첨가한 W/O/W multiple emulsion의 유효성 측정결과 72
1) EGCG를 첨가한 W/O/W multiple emulsion의 항산화능 측정결과 72
2) EGCG를 첨가한 W/O/W multiple emulsion의 미백효과 측정결과 78
3) EGCG를 첨가한 W/O/W multiple emulsion의 주름 효과 측정결과 80
4) EGCG를 첨가한 W/O/W multiple emulsion의 항염증 효과 측정결과 82
8. EGCG를 첨가한 화장품의 안전성 측정결과 84
1) 첩포시험 84
IV. 참고문헌 86
초록 94
Abstract 98
Table 1. The experimental formulation of the W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate(EGCG). 33
Table 2. Check points in stability test of the W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate in constant temperature condition (0, 25, 40℃). 37
Table 3. The expression method of patch test. 40
Table 4. Antimicrobial activity of (-)-epigallo-catechin-3-O-gallate on several microorganisms. 60
Table 5. pH measurement of (-)-epigallo-catechin-3-O-gallate. 64
Table 6. Result of stability test of the W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate in constant temperature conditions(0, 25, 40℃). 69
Table 7. Result of stability test of the W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate in cycle chamber condition 71
Fig. 1. Procedure for polyphenolic compound isolated from Korean green tea. 14
Fig. 2. Structure of (-)-epigallocatechin-3-O-gallate isolated from Korean green tea. 15
Fig. 3. Optimization of cell concentration on macrophage cell(RAW 264.7). 26
Fig. 4. Preparation of the W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate. 34
Fig. 5. Cycle temperature change with day for1 cycle.(-15℃,-10℃, 0℃, -5℃, 0℃, 5℃, 10℃, 15℃, 25℃, 37.5℃, 40℃) 38
Fig. 6. Electron donating ability of (-)-epigallo-catechin-3-O-gallate. 42
Fig. 7. SOD-likeactivity of (-)-epigallo-catechin-3-O-gallat. 44
Fig. 8. Superoxide anion radical scavenging activity of (-)-epigallo-catechin-3-O-gallate. 46
Fig. 9. Inhibition rate of (-)-epigallo-catechin-3-O-gallate on tyrosinase. 48
Fig. 10. Inhibition rate of (-)-epigallo-catechin-3-O-gallate on elastase. 50
Fig. 11. Inhibition rate of (-)-epigallo-catechin-3-O-gallate on collagenase. 52
Fig. 12. Inhibition rate of (-)-epigallo-catechin-3-O-gallate on hyaluronidase. 54
Fig. 13. Effect of (-)-epigallo-catechin-3-O-gallate on the production of nitric oxide in RAW 264.7 cells. 56
Fig. 14. Effects of(-)-epigallo-catechin-3-O-gallate on the iNOS protein expression in Raw264.7 cells. Histogram show the densitometric of iNOS protein normalized to GAPDH. Result are means ± S.D. of triplicate date. 58
Fig. 15. Antimicrobial activity of EGCG on several microorganisms. A : Staphylococcus aureus KCTC 1621 B : Staphylococcus epidermidis KCTC 1917 C : Escherichia coli KCTC 1039 D : Propionibacterium acnes KCTC 3314 61
Fig. 16. Microscopic aspect of the W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate. (1 month afterpreparation) 63
Fig. 17. pH measurement of the W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate 65
Fig. 18. Viscosity change of the W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate 67
Fig. 19. Electron donating ability of W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate 73
Fig. 20. SOD-like activity of W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate 75
Fig. 21. Inhibition rate of W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate on NBT xnathine oxidase 77
Fig. 22. Inhibition rate of W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate on tyrosinase 79
Fig. 23. Inhibition rate of W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate on collagenase 81
Fig. 24. Inhibition rate of W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate on hyaluronidase 83
Fig. 25. Photograph of patch test of the W/O/W multiple emulsion containing (-)-epigallo-catechin-3-O-gallate. 85