Research And Development Of Selenium-enriched Mung Bean Fermentation Broth Mask And A Preliminary Study On The Activity Of Selenium-enriched Dark Tea

Mung bean is a crop rich in protein and flavonoids,and has been widely used in cosmetics.Dark tea is a post-fermented tea containing tea polyphenols,tea pigments and other substances.In this study,the selenium-enriched mung bean and selenium-enriched dark tea grown in Enshi,Hubei,a well-known selenium-enriched region in China,were used as research objects.To explore the whitening,moisturizing and antioxidant activities of selenium-enriched mung bean fermentation broth,and antioxidant and antibacterial activities of selenium-enriched dark tea.The main results are as follows:?1?A selenium content of 0.002 ppm,a total nitrogen content of 18.96 mg/mL,and a?-aminobutyric acid??-GABA?content of 195356.08 nmol/mL were determined in the selenium-enriched mung bean fermentation broth which was produced by microbial fermentation.In the DPPH and ABTS free radical scavenging experiments,the scavenging rates of high concentration selenium-enriched mung bean fermentation broth for DPPH and ABTS free radicals were 96.87%and 99.66%,respectively.In vitro tyrosinase inhibition experiments,the tyrosinase inhibition rate of 40?L selenium-enriched mung bean fermentation broth reached 80.72%.In addition,8%selenium-enriched mung bean fermentation broth could make melanin content decreased to 66.79%in mouse melanoma cells?B16F10?.The results of AQP3 content determination in fibroblasts?HFF-1?showed that,the expression levels of AQP3 gene were 1.86,3.12and 4.23 in HFF-1 cells which were treated with 1.56%,3.13%,and 6.25%selenium-enriched mung bean fermentation broth,compared with the expression level of 1.02 in the blank group,the expression level of AQP3 gene increased significantly.It has been proved that the selenium-enriched mung bean fermentation broth has obvious in vitro antioxidant,whitening and moisturizing effects.?2?The dermal irritation test results showed that surfaces of the four guinea pigs were normal,with no adverse reactions such as erythema,eschar and edema.The results of the acute eye irritation test showed that the eyes of the three rabbits did not show congestion or edema,and the cornea was clear,the iris was normal,there was no eye damage such as edema to the near or half of the eyelid closed.The skin allergy test results showed that there was no erythema,scorching,or edema on the skin of twenty guinea pigs after induction exposure and challenge exposure.Mercury?Hg?,lead?Pb?and cadmium were not determined in the selenium-enriched mung bean fermentation broth,and arsenic?As?was only 0.115 ppm,which was far less than the limit of 2 ppm in the relevant regulations of the?Safety and Technical Standards for Cosmetics??2015?.The above experimental results show that the selenium-enriched mung bean fermentation liquid has no irritation to the skin and eyes,it can be applied to skincare products without safety problems.?3?A mask with selenium-enriched mung bean fermentation broth was provided to thirty-one women aged 18-60 years for using four weeks.At 0,2 and 4 weeks,the indicators of anti-aging such as the number of wrinkles,the volume of wrinkles,the area of wrinkles,and the length of wrinkles,the indicators of skin whitening such as the L*value,b*value,ITA?value,and melanin content,the moisturizing indicators such as the cuticle moisture content of the skin?CORNE?and transdermal water loss rate?TEWL?were determined.The results showed that the number,area and length of wrinkles were significantly reduced;the volume of wrinkles did not change significantly at 2 weeks,while decreased significantly after 4 weeks of use.The L*value and ITA°value,which represent the black and whiteness of the skin,became significantly larger.The melanin content was significantly reduced,while the b*values,which represents the yellowness of the skin,was not significantly different.The long-lasting moisturizing results showed that the CORNE of the subjects was significantly increased,and the TEWL was significantly decreased;the short-term moisturizing test showed that the CORNE was increased.Overall,it has been proved that the selenium-enriched mung bean fermentation broth has excellent in vivo anti-aging,whitening and moisturizing effects.?4?The analysis of the components of the selenium-enriched dark tea extract showed that the contents of selenium,tea polysaccharides,tea polyphenols,and flavonoids were higher,which were 0.18 ppm,24.88%,25.42%and 21.1%,respectively,the content of tea pigments was lower.The 0.32 mg/mL selenium-enriched dark tea extract had 92.73%and 98.43%clearance of DPPH and ABTS free radicals,respectively.The selenium-enriched dark tea extract increased the survival rate of 2?MH₂O₂-stimulated HFF-1 cells from 63.32%to 79.83%at 0.08 mg/mL,besides,the contents of ROS,SOD,and MDA in the H₂O₂-stimulated cells are 38.67%,1.08U/10²Cell and 13.75 nmol/10²Cell respectively,ROS and MDA decreased significantly,and SOD increased significantly after treating the cells with a series of selenium-enriched dark tea extracts.At 0.08 mg/mL,the contents of ROS,SOD,and MDA were24.10%,2.01 U/10²Cell and 9.35 nmol/10²Cell.The results of bacteriostatic experiments showed that the extract had an inhibitory effect on S.aureus and Pseudomonas aeruginosa at 0.4 g/mL,but had no inhibitory effect on E.coli and Candida albicans,indicating the inhibitory effect of the dark tea extract on harmful bacteria is not obvious.The contents of mercury,arsenic,lead and cadmium were 0.004ppm,0.058 ppm,0.82 ppm,and 0.104 ppm,which were all within the limits.The dermal irritation test,eye irritation test and skin allergy test have proved the safety of 2%selenium-enriched dark tea extract.