research
NMN VOLUNTARY INSPECTION INFORMATION
acute oral toxicity test
summary
AS A RESULT OF ADMINISTERING OUR NMN RAW MATERIALS TO MICE AND TESTING FOR TOXICITY, NO TOXICITY WAS OBSERVED EVEN WHEN THE MAXIMUM DOSE WAS ADMINISTERED.
test facility
Japan Food Analysis Center, Tama Research Institute
test purpose
Samples will be tested for acute oral toxicity in mice according to the OECD Guideline for Testing of Chemicals 420 (2001).
test overview
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.
1) Deaths
No deaths were observed during the observation period in either male or female treatment group.
2) General condition
No abnormalities were observed during the observation period in either male or female treatment group.
3) Weight change (Tables -1 and 2)
Weight measurements at 7 and 14 days after administration No difference in body weight values was observed in the test groups of both males and females compared to the control group.
4) Necropsy findings
No abnormalities were observed in all test animals of both males and females at necropsy at the end of the observation period.
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.
1) Deaths
No deaths were observed during the observation period in either male or female treatment group.
2) General condition
No abnormalities were observed during the observation period in either male or female treatment group.
3) Weight change (Tables -1 and 2)
Weight measurements at 7 and 14 days after administration No difference in body weight values was observed in the test groups of both males and females compared to the control group.
4) Necropsy findings
No abnormalities were observed in all test animals of both males and females at necropsy at the end of the observation period.
Whitening, anti-aging, and antioxidant tests
summary
Addition of NMN (Nicotinamide mononucleotide) to cells significantly increased the
□ rate of inhibition of tyrosinase activity compared to the negative control.
□ Significantly increased inhibition of elastase activity compared to the negative control.
The percentage of reactive oxygen species (DPPH) scavenging was significantly increased compared to the negative control.
From the above results, NMN (Nicotinamide mononucleotide) is considered to have potential as a raw material for whitening, anti-aging, and anti-oxidation.
test facility
Clean Test Lab, Inc.
test case
□ Tyrosinase inhibitory activity
□ Elastase inhibitory activity
□ Reactive oxygen species (DPPH) scavenging
test overview
IN AGING HUMAN SKIN, AGE SPOTS, WRINKLES, SAGGING, AND OTHER SIGNS OF AGING ARE OBSERVED. TYROSINASE ACTIVITY IN MELANOCYTES PLAYS AN IMPORTANT ROLE IN MELANIN ACCUMULATION IN THE EPIDERMIS. ELASTASE ACTIVITY, WHICH DEGRADES ELASTIC FIBERS (ELASTIN) PRODUCED BY DERMAL FIBROBLASTS, IS THOUGHT TO BE INVOLVED IN THE LOSS OF SKIN ELASTICITY (FIRMNESS), WHICH IS ONE OF THE CAUSES OF WRINKLES AND SAGGING. IN ADDITION, IT IS SAID THAT ABOUT 80% OF WRINKLES AND SAGGING ARE CAUSED BY PHOTOAGING DUE TO LONG-TERM UV EXPOSURE, WHILE VARIOUS PHENOMENA INDUCED IN THE SKIN BY UV LIGHT ARE KNOWN TO BE MEDIATED BY REACTIVE OXYGEN SPECIES. THEREFORE, THE SCAVENGING OF REACTIVE OXYGEN SPECIES IS AN IMPORTANT MEASURE AGAINST PHOTOAGING. IN THIS STUDY, WE EVALUATED THE TYROSINASE INHIBITORY ACTIVITY, ELASTASE INHIBITORY ACTIVITY, AND REACTIVE OXYGEN SPECIES (DPPH) SCAVENGING ABILITY OF THE TEST SUBSTANCE, AND EXAMINED THE POSSIBILITY OF USING THE TEST SUBSTANCE IN ANTI-AGING COSMETICS.
radiation test
summary
The results of the analysis of radioactive iodine (I-131) and radioactive cesium (Cs-137, Cs-134) in our NMN material showed that none of the radioactive substances were detected.
test facility
Farmers' Federation Food Analysis Center, Inc.
test case
Radioactive iodine I-131
Radioactive cesium Cs-137
Radioactive cesium Cs-134
test overview
The following table 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 "Office 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. The measurements of radioactive iodine (I-131) and radioactive cesium (Cs-137, Cs-134) were conducted using a germanium semiconductor detector based on "Office Communication - Matters to be Considered in Inspections - (Washing with Running Water)" (March 18, 2011). For the determination of nuclides, efficiency calibration using the mathematical efficiency creation program "LabSOCS" was used.
sirtuin test
summary
TWENTY-TWO HUMAN TESTS CONFIRMED THAT 3 MONTHS OF NMN TREATMENT SIGNIFICANTLY ACTIVATED THE SIRTUIN GENE.
test facility
Immunoanalysis Center, Inc.
test case
RNA IS EXTRACTED FROM WHOLE BLOOD AND GENE EXPRESSION LEVELS ARE MEASURED BY QUANTITATIVE RT-PCR.
test overview
RNA is extracted from the collected specimen (2.5 cc of frozen whole blood) and the sirtuin gene expression level is measured by quantitative RT-PCR. The cells in the blood are lysed, RNA is extracted, and cDNA obtained by reverse transcription of the RNA is used as a template for quantitative PCR. The Β-actin gene is used as an internal standard for relative quantification of SIRT1 gene expression. In the test, two cell lines with different SIRT1 gene expression levels are simultaneously measured as controls. The coefficient of variation (CV) of the Ct value (the number of cycles when a certain amount of PCR amplified product is reached), which indicates the variability of replicate samples, is on average about 1%.
Absorption testing by body site
summary
NMN ABSORPTION EFFICIENCY IN THE STOMACH, SUBLINGUAL, AND INTESTINE WAS TESTED BY MEASURING NAD+ LEVELS IN THE BLOOD. AS A RESULT, IT WAS CONFIRMED THAT ABSORPTION EFFICIENCY INCREASED 1.1-FOLD IN THE SUBLINGUAL REGION AND 6-FOLD IN THE INTESTINE, BASED ON GASTRIC ABSORPTION.
test facility
Testing FacilityAdvanced Biomedical Research Institute, Inc.
test case
MEASURE CHANGES IN BLOOD NAD+ BEFORE AND AFTER NMN INGESTION FROM THE STOMACH, SUBLINGUAL, AND INTESTINAL SITES.
test overview
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 IS COLLECTED, 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 TOTAL AMOUNT OF NAD+/NADH.
