Protheragen-ING offers comprehensive cosmetic analysis services to ensure the safety and efficacy of cosmetic products. Eye irritation is an important issue for cosmetic products because they come into direct contact with the sensitive area around the eyes.

We offer Hen's Egg Test-Chorioallantoic Membrane (HET-CAM) test services to evaluate the ocular safety of cosmetic products before they are placed on the market, providing our clients with valuable information on regulatory compliance and consumer safety.

What Is the HET-CAM Test?

One of the important tests carried out by Protheragen-ING is the HET-CAM test. The test is an in vitro experiment that evaluates the potential ocular irritation of cosmetic formulations and components using egg embryos.It focuses on evaluating these compounds' potential for causing ocular discomfort by looking at alterations in the egg's chorionic allantoic membrane (CAM).

HET-CAM has been shown to be a robust qualitative method for assessing the potential irritation of chemicals. The CAM is an entire tissue with veins, arteries, and capillaries. It reacts to damage by inducing an inflammatory reaction like to that seen in the conjunctiva of rabbits. Its highly developed vascularization makes it the perfect model for research on ocular irritation.

The hen egg chorioallantoic membrane.Fig. 1 The hen's egg chorioallantoic membrane. (McKenzie B, et al. 2015)

Our HET-CAM Testing Services

Protheragen-ING offers comprehensive HET-CAM testing services to meet your specific needs. We follow standardized protocols and use state-of-the-art equipment to provide accurate and reliable results. Below is an overview of the services we offer:

Sample Preparation

Protheragen-ING will assist you in preparing your test samples. We ensure that the samples are properly formulated and adjusted to the concentration required for accurate evaluation.

HET-CAM Test Procedure

We carefully open the incubated eggs and expose the CAM on day 9 and place the test sample in pure or diluted solution directly on the exposed chorionic urothelium membrane and visually inspect the membrane with a microscope for any signs of irritation or damage.

The reaction time on the membrane was 5 minutes. We carefully record and analyze the test results to provide you with a thorough evaluation.

Schematic representation of the chorioallantoic membrane vascular assay (CAMVA), also known as the Hen egg test (HET).Fig. 2 Schematic representation of the chorioallantoic membrane vascular assay (CAMVA), also known as the Hen's egg test (HET). (Wilson SL, et al. 2015)

Reactions result in bleeding, coagulation and fragmentation. Based on the reaction time, an irritation score is calculated and the items are categorized according to the score as follows.

0 to 0.9 points Non-irritating
1 to 4.9 points Slightly irritating
5 to 8.9 points Moderately irritating
9 to 21 points Strongly irritating

Data Analysis and Reporting

Upon completion of the HET-CAM test, Protheragen-ING thoroughly analyzes the data obtained. We generate a detailed report that summarizes the test results, including any observed irritation effects, scores, and interpretation of the results.

Our reports are clear and concise and are designed to help you make informed decisions about the safety of your cosmetic products.

Contact Protheragen-ING today to discuss your needs or to learn more about our comprehensive cosmetic analysis solutions.We will respond within 2-4 working days.


  1. Wilson SL, et al. (2015). "An Overview of Current Techniques for Ocular Toxicity Testing." Toxicology, 327, 32-46.
  2. McKenzie B, et al. (2015). "The hen's egg chorioallantoic membrane (HET-CAM) test to predict the ophthalmic irritation potential of a cysteamine-containing gel: Quantification using Photoshop® and ImageJ." International Journal of Pharmaceutics, 490(1-2), 1-8.


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