Skin-whitening Efficacy and Dosage Form Effect of Modified Seven-White Chinese Herbal Formulation

• Main Authors: Shin-Yih Rao, 饒欣宜
• Other Authors: Jyi-Faa Hwang
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/81042117148503726886

碩士 === 大仁科技大學 === 生物科技研究所 === 101 === The objective of this study was to identify efficient skin-whitening formulation and dosage forms by using the traditional seven-white Chinese herbal formulation as the starting basis, then add or remove individual ingredients by screening their antioxidant and tyrosinase inhibitory activity. On top of that, oligopeptides and extract of natural plant, quercetin, were also included in the formulation for preventing and repairing possible photodamages associated with skin whitening. A secondary objective of this study was to establish a feasible platform for testing the clinical efficiency of skin-whitening cosmetic formulation. The experimental work of this study consisted of three parts: First, the screening of individual and grouped ingredients for free radical scavenging and tyrosinase inhibiting activity. Selected formulae with grouped ingredients showing extraordinary antioxidant and anti-tyrosinase activity were also studied for tyrosinase kinetic parameters to determine their mechanism of inhibition. Second, the skin sensitining test and skin-whitening efficacy test of dosage forms based on modified seven-white Chinese herbal formulation. Third, the treatment of B16 melanoma cells with selected formulae for testing their growth retardation activity and their inhibitory activity against melanin synthesis within B16 melanoma cells. The results showed that, among extracts of the modified seven-white formulae, selected based on their combined performance of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity and tyrosinase inhibitory activity, the 25% and 50% ethanol extracts of formulae D5, D6 and D7 demonstrated the best skin-whitening effect. The tyrosinase kinetic parameter of D7-50 (50% ethanol extract of D7 formulations, weight ratio of Atractylodes macrocephala: Paeonia lactiflora: Taxillus chinensis : White Rose = 10:3:3:3) revealed a Michaelis Menten constant KM of about 0.02 mM, much smaller than 0.275 mM of α-Arbutin, indicating that the affinity of D7-50 extract with tyrosine is considerably good. Furthermore, the pattern of its Michaelis Menten plot, compared with that without tyrosinase inhibitor, suggests that D7-50 is a non-competitive inhibitor of tyrosine and tyrosinase. On the skin of 10 volunteers, and 6 Specific Pathogens Free mice, that applied the cream base of this study containing extracts of D5, D6 and D7 formulae, no significant skin sensitization phenomena was observed. Among 10 volunteers, age 25 to 30, that applied facially the cream made of quercetin and oligopeptide-containing D7-50 formula for four consecutive weeks, more than 50% showed apparent skin-whitening effect based on Fitzpatrick’s skin typing. During preliminary test, the treatment of B16 melanoma cells with quercetin (Q)-containing Q+D7-50 extract showed considerable efficacy in retarding the growth of B16 melanoma cell, and in suppressing melanin synthesis within B16 melanoma cells. For instance, the Q+D7-50 extract (1:9; 0.02 mg/mL) demonstrated inhibitory effect equivalent to 80% that of the control standard vitamin C (0.2 mg/mL) against melanin synthesis within B16 melanoma cells. This seems to suggest that the utilization of B16 melanoma cells as a platform to evaluate the skin-whitening efficiency of modified seven-white Chinese herbal formulation containing non-conventional ingredients, such as quercetin and oligopeptides, is feasible.