Irritation Test

There was no damage on the surface of the chicken chorioallantoic membrane (HET-CAM), and there was no irritation of blood vessels, blood clotting or melting. According to the scoring criteria, ES=0 for no eye irritation.

Irritation tests
Cell culture tests and human skin tests
Cell cultures are used to check whether a product poses a definitive hazard to normal human cells after penetration into the skin (cell mortality >50%).

Before the human test, BioTruly will conduct an internal risk evaluation of the raw materials or formulation’s ingredients to confirm that there is no risky substance in excess of the hazard components. Their pH value should be within a reasonable range so that we can conduct human test. We use the gold standard cosmeceutical evaluation method: PATCH TEST. Patches are applied directly to the surface of human skin for 24-48 hours. It is used to confirm whether the product will have a clear irritating effect on normal skin when it leave on the body for a long time.

In the future, BioTruly will continue to pay attention to more alternative testing methods. In order not to harm animals, we will use high quality and reliable models to proceed cosmetics irritation tests.

Safety Evaluation
Result of Eye irritation test
The eye irritation method of BioTruly fermented serum (ie, the original cosmetic shown in the chart) was evaluated by qualitative and quantitative methods using in vitro experiments: HET-CAM experiments and CAM-TBs.

Result of HET-CAM test
After the reaction of sample and the chicken chorioallantoic membrane, the blood vessels on the surface of the allantoic membrane were hemorrhagic, coagulated, dissolved, and graded to evaluate the intensity of eye irritation.
The judgment criteria and test methods are shown in Annex 1, and the test results are shown in Figure 1.

Before use, the surface of the chicken embryo chorioallantoic membrane was not damaged. The blood vessel was intact without damage, which met the experimental requirements.
After use, the surface of the chicken embryo allantoic membrane was not damaged. The blood vessels did not have the irritation of bleeding, coagulation, and melting.
According to the scores, BioTruly fermented serum resulted to be no eye irritation. Through HET-CAM experiment, ES=0.

Result of CAM-TBs test
After the sample reacts with the chicken embryo chorioallantoic membrane, the damaged allantoic membrane can adsorb the blue pepper. The sample eye irritation was evaluated by quantitatively measuring the amount of the mixture. The judgment criteria and test methods are shown in Annex 2. The specific test results are shown in Figure 2.

According to the negative control and positive control results, the test method and process are credible.
According to the scoring standard, the absorption of the Trypan Blue of fermented serum is less than 15 nmol. So no irritation.

Skin Model Test
The skin irritation was determined by the size of epidermal cell activity in the skin model after the sample reacted with the skin model. The specific criteria and test methods are shown in Annex 4. The specific results are shown in Figure 4.

According to the blank control, negative control, and positive control results, the test process and method are credible and correct.
According to the judgment criteria, the cell viability of BioTruly fermented serum was greater than 50%, indicating that it did not cause skin irritation.

PATCH TEST
BioTruly fermented serum are applied for patch test. Only one of the 30 volunteers had a first grade reaction when the patch was removed after 0.5 h. There was weak redness reaction occurred in the test area. The specific results are shown in Figure 5. The volunteers’ response record is shown in the appendix. According to the standard, if the number of people whose response is Level 1 less than 5 people, it results that sample did not cause negative reactions on human body and not irritation.