| With the acceleration of the aging process,people’s anti-aging demand is increasing,so it is of great significance to develop products with anti-aging effect.A large number of studies have proved that both Nicotinamide Mononucleotide(NMN)and blueberry have anti-aging effects,but the most common NMN products on the market currently remain in the form of capsules,tablets and other relatively simple products,so this paper developed a liquid drink with anti-aging effects,which not only enriches the product form,but also conforms to the market development.The main research results are as follows:(1)NMN blueberry liquid drink was prepared with NMN,blueberry and white granulated sugar as raw materials.The optimal conditions of complex enzymolysis were obtained by single factor and orthogonal enzymolysis tests with the juice yield as the standard: the addition of pectinase was 0.5%,the addition of cellulase was 1.2%,the enzymolysis time was 2.0 h,and the enzymolysis temperature was 55℃.The supernatant of enzymolysated blueberry pulp was centrifuged for color protection experiment.The optimal color protection agent was 0.4 g /L ascorbic acid based on the Browning degree and anthocyanin content.On this basis,the best processing technology of NMN blueberry liquid drink was obtained,and the liquid drink was good in flavor,rich in taste and stable in color.(2)The liquid drink was prepared according to the final processing technology.The physical and chemical indexes and main components of the liquid drink were measured and calculated according to the national standards and the common methods for the determination of physical and chemical indexes,such as ultra-high performance liquid chromatography tandem mass spectrometry,p H differential method,and Folinphenol method.Finally,the content of NMN in NMN blueberry liquid drink was 6.01mg/mL.Anthocyanin content was 157.30 mg/L;Polyphenol content was 0.0459 μg/mL;Flavonoid content was 1.78 g/L;Browning degree OD value was 0.321;Soluble solid content was 12.47 Brix%;pH was 3.51;Protein content was 0.42 g/100 g;Fat content is 0.40 g /100 g;Carbohydrate content is 9.70 g /100 g;The energy is 187 KJ/100 g;Sodium content is 166 mg/100 g;Water content was 89.2%;Ash content was 0.28 g/100 g;Dietary fiber was not detected.(3)The prepared NMN blueberry liquid drink was stored at -18℃,4℃ and 25℃in the dark to study the quality changes of liquid drink during storage.The results showed that with the increase of storage temperature and storage time,anthocyanin content and NMN content in liquid drinks showed a decreasing trend,but the decrease rate of anthocyanin content and NMN content was slow at -18℃ and 4℃,and the decrease trend of NMN content gradually became gentle with the increase of storage time.The higher the temperature is,the greater the Browning degree is,and the lower the Browning degree is at low temperature.However,solid content and pH value did not change significantly during storage,and no bacteria was detected in NMN blueberry liquid drink at three temperatures during storage.It is concluded that low temperature storage is beneficial to the preservation of nutrients and color protection of NMN blueberry liquid drink.However,because -18℃ may cause trouble in thawing and microbial contamination in the thawing process,4℃ is a better storage temperature.(4)Stable conditions were found through D-galactose concentration pretest,and metformin was used as positive control.ROS staining and β-galactosidase staining were used to verify the successful establishment of the aging model.The results showed that 40 mg/mL D-galactose could induce the production of reactive oxygen species and improve the activity of β-galactosidase,a marker of aging,in zebrafish compared with the blank control group and the positive control group.In addition,the zebrafish aging model experiment established under this condition requires short time,is simple to operate,and the effect is stable,which provides a new idea for the establishment of zebrafish aging model with D-galactose.(5)The anti-aging effect of NMN Blueberry liquid drink was verified by establishing a successful aging model of zebrafish.By comparing the ROS level,MDA content,CAT enzyme activity,SOD enzyme activity and β-galactosidase activity of zebrafish larvae in different experimental groups,it was found that NMN blueberry liquid drink could inhibit the generation of D-galactose induced reactive oxygen species in zebrafish,and reduce the content of lipid peroxidation product MDA in zebrafish larvae induced by D-galactose.It can increase the activities of CAT enzyme,SOD enzyme and β-galactosidase in zebrafish.The above results indicate that NMN blueberry liquid drink has certain anti-aging activity. |
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