표제지
목차
Ⅰ. 서론 15
Ⅱ. 재료 및 방법 18
1. 실험 재료 18
(1) 재료 18
(2) 시료 추출 18
(3) 초임계를 이용한 추출 20
(4) Column chromatography 를 이용한 미백활성물질의 분리 21
(5) 시약 및 기기 23
2. 실험 방법 26
(1) 항산화 효과 측정 26
1) 전자공여능 측정 26
2) Superoxide dismutase (SOD) 유사활성 측정 27
3) Xanthine oxidase 저해활성 측정 28
(2) 미백 효과 측정 29
1) Tyrosinase 저해 활성 측정 29
2) Column chromatography 로 분리된 분획의 Tyrosinse 저해측정 29
(3) Astringent 활성측정 31
(4) 아질산염 소거능 측정 32
(5) 항염효과 측정 34
1) 판람근 추출물에 의한 Macrophage cell 의 생존율측정 34
2) Nitric oxide 저해활성 측정 35
3) Western blot 에 의한 iNOS 단백질 발현량 측정 36
(6) 항암 효과 측정 39
1) 세포배양 39
2) MTT assay 에 의한 항암 효과 측정 39
(7) PCR 에 의한 Propionibacterium acnes 동정 41
(8) 항균 효과 측정 44
1) 균배양 44
2) 생육 저해환 (Clear zone) 측정 44
(9) 통계처리 46
(10) 판람근 크림의 안정성 및 안전성 실험 47
1) 판람근 크림의 제조 처방 및 방법 47
2) 판람근 크림의 색도 측정 47
3) pH 측정 49
4) 점도 측정 50
5) 동적 점탄성 측정 (Oscillatory shear test) 51
6) 일반적 보존 시험 53
가. 온도 (Incubation) 에 따른 안정성 시험법 53
나. 광안정성 시험법 54
a. 인공광 (Artificial sun lamp) 노출 시험법 54
b. 자연광 노출 시험법 54
7) 특수ㆍ가혹 보존 시험 55
가. 온도순환 (Cycle chamber) 에 따른 안정성 시험법 55
8) 판람근 크림의 안전성 시험 56
(1) 판람근 크림의 안정성 측정결과 94
1) 판람근 크림의 제조 94
2) 색도 측정결과 94
3) 판람근 추출물 및 판람근 크림의 pH 측정결과 97
4) 판람근 크림의 점도 측정결과 101
5) 판람근 크림의 동적 점탄성 (Oscillatory shear test) 측정결과 104
가. 복소탄성률 (Complex modulus, G) 측정결과 104
나. 손실각 (Loss angle, δ ) 측정결과 105
6) 일반적 보존 시험 108
가. 온도 (Incubation) 에 따른 안정성 결과 108
나. 광안정성 110
a. 인공광 (Artificial sun lamp) 노출 결과 110
b. 자연광 노출 결과 110
7) 특수ㆍ가혹 보존 시험 113
가. 온도순환 (Cycle chamber)에 따른 안정성 시험결과 113
(2) 판람근 크림의 안전성 측정 115
1) 첩포 시험결과 115
Ⅳ. 참고문헌 117
초록 130
Abstract 132
Table 1. Condition of Isatis tinctoria L. supercritical fluid extraction 20
Table 2. The experimental formulation of the cream containing Isatis tinctoria L. 48
Table 3. Check points in stability test of the cream containing Isatis tinctoria L. in constant temperature conditions (0, 25, 40 ) 53
Table 4. The expression method of patch test 56
Table 5. Antimicrobial activity of Isatis tinctoria L. on several microorganisms 91
Table 6. Changes in the Hunter color values of cream containing Isatis tinctoria L. 91
Table 7. Results of stability test of the cream containing Isatis tinctoria L . in constant temperature conditions 109
Table 8. Results of Artificial Sun lamp test of the cream containing Isatis tinctoria L. 111
Table 9. Results of Sun test of the cream containing Isatis tinctoria L. 112
Table 10. Results of stability test of the cream containing Isatis tinctoria L. in cycle chamber condition 114
Table 11. Results for patch test of the cream containing Isatis tinctoria L. 116
Fig. 1. Procedure for extraction from Isatis tinctoria L. 19
Fig. 2. Procedure for methanol extraction from Isatis tinctoria L. 21
Fig. 3. Procedure for CHCl₃/MeOH fraction form /Isatis tinctoria L . 22
Fig. 4. Photograph for Thin layer chromatography form Isatis tinctoria L. extracted . 22
Fig. 5. The method of electron donating ability. 26
Fig. 6. The method of superoxide dismutase (SOD)-like activity. 27
Fig. 7. The method of xanthine oxidase inhibition. 28
Fig. 8. The method of tyrosinase inhibition. 30
Fig. 9. The method of Astringent activity. 31
Fig. 10. The method of Nitrite-scavenging ability. 33
Fig. 11. The method of cell viability on Raw 264.7 34
Fig. 12. The method of NO - reduction. 35
Fig. 13. The method of Western blot. 37
Fig. 14. Western blotting antibody reaction. 38
Fig. 15. The method of growth inhibition on cancer cell. 40
Fig. 16. Primers used for the amplification of 16S rRNA of Propionibacterium acnes acnes. 42
Fig. 17. The polymerase Chain Reaction. 43
Fig. 18. The identification method of inhibition zone on several microorganism. 45
Fig. 19. Procedure of viscosity measurement. 50
Fig. 20. Procedure of rheological properties measurement. 52
Fig. 21. Cycle temperature change with day for 1 cycle. 55
Fig. 22. Electron donating ability of Isatis tinctoria L. extracted. 59
Fig. 23. SOD-like activity of Isatis tinctoria L. extracted. 62
Fig. 24. Inhibition rate of Isatis tinctoria L. extracted on xanthine oxidase. 64
Fig. 25. Inhibition rate of Isatis tinctoria L. extracted on tyrosinase. 67
Fig. 26. Inhibition rate of the various fractions from Isatis 68
Fig. 27. Comparison of astringent activities of Isatis tinctoria L . 71
Fig. 28. Nitrite scavenging ability of Isatis tinctoria L . 73
Fig. 29. Viability of Isatis tinctoria L. extract on Macrophage cell (RAW 264.7). 75
Fig. 30. Inhibition rate of Isatis tinctoria L. ethanol extract on Nitric oxide production. 77
Fig. 31. Inhibition rate of Isatis tinctoria L. water extract on Nitric oxide production. 78
Fig. 32. Modulation by the Isatis tinctoria L. ethanol extract of LPS-induced iNOS expression in RAW 264.7 cells. LPS-induced iNOS expression in RAW 264.7 cells. 80
Fig. 34. Growth inhibition rate of Isatis tinctoria L. extract on mouse melanoma cancer(B16F10). 84
Fig. 35. Growth inhibition rate of Isatis tinctoria Linne extract on breast cancer(MDA). 85
Fig. 36. Growth inhibition rate of Isatis tinctoria L. extract on lung cancer(A549). 86
Fig. 37. Growth inhibition rate of Isatis tinctoria L. extract on colon cancer(HT-29). 87
Fig. 38. PCR products the strains tested for 16s rRNA of Propionibacterium acnes. 89
Fig. 39. Antimicrobial activity of Isatis tinctoria L. with supercritical fluid extracted on several microorganisms. 92
Fig. 40. Antimicrobial activity of Isatis tinctoria L. with supercritical fluid extracted on several microorganisms. 93
Fig. 41. Photograph of the cream containing Isatis tinctoria L. extracted. 95
Fig. 42. pH change with concentration of of Isatis tinctoria L. 98
Fig. 43. pH change of the cream containing Isatis tinctoria L . extracted with water water. 99
Fig. 44. pH change of the cream containing Isatis tinctoria Linne extracted with ethanol ethanol. 100
Fig. 45. Viscosity change of the cream containing Isatis tinctoria L. extracted with water. 102
Fig. 46. Viscosity change of the cream containing Isatis tinctoria L. extracted with ethanol. 103
Fig. 47. Complex modulus change with concentration of the cream containing Isatis tinctoria L. 106
Fig. 48. Loss angle changes of the cream containing Isatis tinctoria L. 107