Validation Study in Korea for HET-CAM Test and NRU Assay as Alternatives to the Draize Eye Irritation Test

Bae-Hwan Kim1, Seong-Joon Moon1, Ih-Seop Chang1, Young-Chul Sim1, Jin-Cheon Hong2, Ki-Moon Kim3, Moon-Uk Park4, Chang-Seok Rhoo5 and Min-Seok Jeong6
1 Skin Research Institute, Pacific Co. R&D Center, Kyonggi-do 449-729, Korea
2 Nadri Cosmetic Co., Kyonggi-do 451-860, Korea
3 Boryung Pharm. Co., Kyonggi-do 435-050, Korea
4 LG Chem. Research Park, Taejon 305-343, Korea
5 Coreana Cosmetic Co. Ltd., Chungchung 333-830 Nam-do, Korea
6 Hankuk Cosmetic Co. Ltd., Kyonggi-do 421-150, Korea
Correspondence: Bae-Hwan Kim, DVM
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Original paper :AATEX 8(3-4):78-90
The objective of this study is the validation of two alternative methods to rabbit eye irritation test (Draize test). The henfs egg chorioallantoic membrane (HET-CAM) test is actively studied by several countries as an alternative to the rabbit eye irritation test. It employs the vascular membrane of a fertile henfs egg to assess eye irritation potential. We assembled a co-operating organization including 6 cosmetic research centers and carried out validation studies to assure the HET-CAM test as an acceptable alternative to the Draize test. The NRU assay, a cytotoxicity test was also conducted in this study to complement the HET-CAM method.
We selected 25 substances for pre-validation study, 20 substances for 1st- validation study, and 12 substances for 2nd-validation study from common cosmetic ingredients. The correlation study to compare HET-CAM test, NRU assay and Draize test was performed with Pearson and Spearman correlation coefficient values. We also analyzed the inter-laboratory correlation based on the results of the HET-CAM obtained by 6 cosmetic research centers.
We conclude that the HET-CAM test and the NRU assay are good methods to replace the Draize test because of the high correlations observed among HET-CAM test, NRU assay and Draize test. However, additional validation studies are needed to accept these methods as integral components of the guideline due to the big inter-laboratory validation. Keywords: Validation study, in vitro alternatives, Draize test, HET-CAM, NRU assay, cytotoxicity