Five-week-old male and female ICR mice were pre-reared for approximately one week to ensure that there were no abnormalities in their general condition before being used for the study. The test group was administered 2000 mg/kg as the specimen dose, and the control group was administered injectable water as the solvent control.
The test animals were fasted for approximately 4 hours prior to administration. After weighing the animals, a forced single oral dose of the test solution was administered to the test group and a forced single oral dose of water for injection was administered to the control group at a dose volume of 20 mL/kg, respectively, using a gastric sonde.
The observation period was 14 days, with frequent observation on the day of administration and once a day from the following day. Body weights were measured at 7 and 14 days after administration, and Levene's test was performed. Since there was no difference in variance, Student's t-test was used to compare the groups. The significance level was set at 5%. All animals were necropsied at the end of the observation period. The following findings were observed:

1) No deaths were observed in either sex during the observation period.

2) No abnormalities were observed in either sex during the observation period.

3) Weight changes (Tables - 1 and 2) Weighing at 7 and 14 days after administration No difference in body weight was observed in the test groups of both males and females compared to the control group.

4) Necropsy findingsAt necropsy at the end of the observation period, no abnormalities were observed in all test animals of both males and females.

The following is based on the "Manual for Measuring Radioactivity of Foods in Emergency Situations" (March 2002, Monitoring and Safety Division, Food Insurance Department, Pharmaceutical Affairs Bureau, Ministry of Health, Labour and Welfare), "No. 7 Gamma-ray Spectrometry with Germanium Semiconductor Detector" (revised 1992), Radioactivity Measurement Method Series, Ministry of Education, Culture, Sports, Science and Technology (MEXT), and "Administrative Communication - Points to be noted in inspections - (washing with running water)" (March 18, 2011), Monitoring and Safety Division, Food Safety Department, Pharmaceutical Affairs Bureau, Ministry of Health, Labour and Welfare. Based on the "Office Communication - Matters to be noted in inspections - (washing with running water)" (March 18, 2011), measurements of radioactive iodine (I-131) and radioactive cesium (Cs-137, Cs-134) were conducted using a germanium semiconductor detector. For quantification of nuclides, efficiency calibration with the mathematical efficiency creation program "LabSOCS" was used.

DETERMINATION OF TOTAL NAD+/NADH, NADH AND NAD+ AMOUNTS IN SERUM AND THE RATIO OF NAD+ TO NADH. BY HEAT TREATMENT OF CELL LYSATES PREPARED WITH EXTRACTION BUFFER, ONLY THE AMOUNT OF INTRACELLULAR NADH CAN BE QUANTIFIED, AND THE AMOUNT OF INTRACELLULAR NAD+ CAN BE OBTAINED BY SUBTRACTING THE AMOUNT OF NADH FROM THE SEPARATELY MEASURED TOTAL NAD+/NADH AMOUNT. AFTER BLOOD COLLECTION, SERUM IS SEPARATED, PROTEINS ARE REMOVED BY ULTRAFILTRATION, AND THE TOTAL NAD+/NADH CONTENT IS MEASURED BY THE COLORIMETRIC METHOD. SINCE NAD+ IS DEGRADED BY HEAT TREATMENT, ONLY THE AMOUNT OF NADH IN THE SERUM CAN BE DETERMINED. THE AMOUNT OF NAD+ CAN BE CALCULATED BY SUBTRACTING THE AMOUNT OF NADH FROM THE AMOUNT OF TOTAL NAD+/NADH.

This study was conducted as a single-blind, non-application controlled comparative study to evaluate the anti-wrinkle effect of NMN serum application on the skin. If they are using mosaic, eye tape, or other products to form double eyelids, they must be able to remove the products when they come to the center, The 23 subjects who fulfilled the selection criteria of having the same level of difference in wrinkle grade between the left and right eye corners (up to a difference of 1.0 was acceptable) were subjected to a wrinkle grade visual judgment (Trained Expert), digital photography (anti-wrinkle test), and wrinkle replica analysis (3D wrinkle analysis) according to the ethical principles based on the Declaration of Helsinki. analysis) were performed.
In the wrinkle grade visual inspection, the wrinkle grade of the treatment group was significantly lower than that of the non-application group after 4 weeks of treatment in the wrinkle grade evaluation item.
In the visual wrinkle grade evaluation, the wrinkle grade of the NMN-applied group was significantly lower than that of the non-applied group after 4 weeks of application in the wrinkle grade evaluation item, where a smaller value is better. Therefore, the anti-wrinkle evaluation of the skin by the application of the NMN-formulated serum was found to be effective.

A CLOSED PATCH TEST (OBSERVATION PERIOD: 3 DAYS) WAS CONDUCTED ON 20 JAPANESE MALE AND FEMALE SUBJECTS, AND THE NUMBER OF SKIN IRRITATIONS IN THE CLOSED PATCH TEST USING CONTINUOUS APPLICATION FOR 24 HOURS WAS 0.0 FOR THE SERUM CONTAINING NMN, INDICATING THAT IT WAS A SAFE PRODUCT.