| Whitening and moisturizing are the skin care demands which consumers always be keen to. Application of natural, safe and effective plant extracts has become a hot spot of cosmetics development. Yellow and blue whole flower of different concentrations of ethanol extracts and ovary extract of Nymphaea odorata Aiton as the research objects, determining the main active ingredients and comparing the antioxidant capacity. Through in vitro assays, comparing tyrosinase activity inhibitory effect and moisturizing property of the extracts.Then screening extracts with good effects, through cell culture model using indicators such as intracellular tyrosinase activity and melanin content to evaluate the whitening effect. Through human trials using indicators such as skin moisture content and water loss rate to evaluate the moisturizing effect. The main results are as follows:(1) Determination of total phenols, flavonoids and anthocyanins content in Nymphaea odorata Aiton extracts by colorimetric. Total phenols and anthocyanins content in yellow and blue whole flower with 60% alcohol extracts were more than other extracts. Total phenols content were 30.29±0.19%,31.62±0.14%, flavonoids content were 5.37±0.05%,3.72±0.06% and anthocyanins content were 0.076±0.005%, 0.089±0.009%. The main components of ovary extract were polysaccharides, protein and amino acids. Polysaccharide content was 13.22±0.05%, total carbohydrate content was 35.1±1.14% and the crude protein content was 8.17±0.02%. (2) Scavenging H2O2,O2-· and HO· ability were measured by chemiluminescence and FRAP total antioxidant capacity was measured by spectrophotometry. The results showed that the whole flower extracts had good antioxidant effect. The effect of blue whole flower extracts were stronger than yellow whole flower extracts, of which 60% ethanol extracts were the strongest. The IC50 values of scavenging H2O2 of yellow and blue 60% alcohol extracts were 0.38±0.01 ?g/mL and 0.35±0.02?g/mL and blue 60% alcohol extract was significantly better than Vc (0.55±0.04?g/mL) (P<0.01). The IC50 values of scavenging O2-· were 5.21±0.31?g/mL and 4.53±0.07?g/mL, which were better than Vc (50.12±0.59 ?g/mL) (P<0.01). The IC50 value of scavenging HO· of yellow whole flower with 60% alcohol extract (1.73±0.04?g/mL) was weaker than Vc (0.68±0.03?g/mL), while the blue whole flower extract (0.50±0.09?g/mL) was similar with Vc (P>0.01).(3) In the systems of scavenging HO· and total antioxidant capacity assayed by FRAP, phenols content and antioxidant effect were positive correlation of 0.8814 and 0.8698 respectively. Therefore, presumably polyphenols is one of the main active substances and flavonoids enhances the effect.(4) Results of inhibiting tyrosinase activity in vitro showed that 60% alcohol extracts were the strongest. For further studies, using mouse B16 melanoma cell model to evaluate the whitening effect. The results showed that yellow and blue whole flower with 60% alcohol extracts could effectively inhibit the proliferation of melanoma cells and reduce melanin synthesis. At 100?g/mL concentration, cell proliferation rate decreased to 84.44±1.77% and 48.24±3.14%, relative melanin content reduced 30.31% and 33.02%, and they could effectively inhibit tyrosinase activity in cells, which indicating that they had an effective whitening effect.(5) In vitro moisture absorption experiments showed that the moisture absorption rate and moisturizing rate of water extracts were better than alcohol extracts. Moreover, blue flower extracts were better than yellow flower extracts. Water loss rate of ovary extract was lower than yellow and blue whole flower water extracts at the same concentration. The water loss rate of 1% concentration of ovary extract solution at 12th hour was significantly lower than the same concentration controls of hyaluronic acid (14.20±0.16%) (P<0.01) and it was similar with glycerin (13.49±0.11%).Select ovary extract and blue whole flower water extract for further human trials. The results showed that after using the samples, the relative moisture content of human skin was significantly increased as well as relatived to the control group (P<0.01). After 4 h, the MMV values were 37.28±6.36% and 37.71±7.09% and there were no significant difference compared with hyaluronic acid (36.58±8.27%) (P>0.05). Skin water loss rate was significantly reduced compared to the previous use (P<0.05), which were significantly better compared to the control group after 4 h (P<0.01). Skin elasticity values of R2 and R7 also had improvement. Thus, the ovary extract and blue flower water extract had good moisture capacity.To sum up, Nymphaea odorata Aiton extracts have good antioxidant activity. Whole flower with 60% alcohol extracts can reduce melanogenesis to make skin white. The whole flower water extracts and ovary extract have moisturizing effect. As a new kind of whitening moisturizer, it has a broad application prospects in cosmetics. |
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