| 摘要: | 傳統的化粧品安全性評估方法是以小鼠或兔子作為受測動物，但歐盟已從2009年3月起禁止利用動物進行化粧品的測試，並且歐盟化粧品銷售的法規從2013年起更規定禁止販售以利用動物作為毒性安全實驗的化粧品，因此各大化粧品公司及學術單位都積極地投入非動物型粧品毒性的安全檢測方法，而目前已發展的方法，主要是以細胞毒性檢測作為化粧品安全的指標。但仍有以非細胞毒性的檢測方法做為化粧品安全性的評估，如量測可被化粧品原料調控參與生理代謝過程中的特定酵素活性及觀察化粧品原料對穩定生物分子中二、三和四級結構的化學鍵能 (氫鍵) 的影響，因為這些酵素活性及化學鍵能的破壞將終止許多生理上重要的代謝過程，進而造成人體的毒性。本研究所發展的非動物性化粧品安全性評估策略正是建立在瓊脂凝固的過程中，化粧品原料對化學鍵能的影響。瓊脂是由海藻中提取的多醣體，其降溫凝固的過程中與氫鍵的形成有密切的關係，本實驗選用瓊脂的凝固作為化粧品安全性評估的模式，探討額外添加的化粧品原料對瓊脂凝固的影響並進行毒性的評估。為了解化粧品原料與瓊脂凝固程度的關係，本研究評估化粧品原料對瓊脂凝固時間的影響，探討其與化粧品原料的細胞毒性IC50及小鼠口服急毒性LD50之間的相關性，並以正確率、敏感性、特異性與化粧品安全檢測套組做對照比較。結果顯示瓊脂的凝固實驗結果與小鼠口服急毒性LD50相比，其正確率為73.6%，敏感性為43.7%，特異性為95.4%，偽陽性為23.6%，偽陰性為2.6%；與B16-F10的細胞毒性實驗結果相比，其正確率為84.2%，敏感性為60%，特異性為100%，偽陽性為15.7%，偽陰性為0%；與3T3的細胞毒性實驗結果相比，其正確率為89.4%，敏感性為73.3%，特異性為100%，偽陽性為10.5%，偽陰性為0%。此外瓊脂的凝固實驗結果與化粧品安全檢測套組的結果相比，瓊脂凝固結果具有較低的偽陰性。由以上結果分析，瓊脂的凝固實驗具有作為非動物性化粧品安全性評估檢測方法的潛力並可降低動物實驗的使用量。
The experimental testees of traditional cosmetics safety evaluation are mainly mice or rabbits. As of March 2009, the EU forbade the use of animals in cosmetics testing. Starting from 2013, the EU further forbade the sales in the EU region of cosmetics that used animals in toxicity tests. Therefore, various leading cosmetics companies and academic institutions are actively engaged in developing non-animal safety testing methods. Currently, the methods mainly involve the use of cells to test toxicity. There are also other non-animal methods that test whether the cosmetics will restrain some enzyme activities or change the active macromolecular structure, resulting in cellular degeneration. These methods are like this experiment, using non-living methods to test toxicity. Hydrogen bonding is an integral bonding in the composition of an organism. A substance that damages hydrogen bonds may also damage or prevent the formation of the macromolecular structure, which is toxic to human bodies.But there is still a method to detect non-cytotoxic as cosmetics safety assessment, as measured cosmetic ingredients may be physiological processes involved in the regulation of enzyme activity and observe specific cosmetic ingredients for stable biomolecules II, III and IV bond energy (hydrogen) impact structure because the enzyme activity and destruction of these chemical bonds will be able to terminate a number of important physiological metabolic processes, thuscausing the toxicity. Non-animal cosmetics safety assessment strategy of the Institute for Development is built on the agar solidification process, the impact on the bond energy of cosmetic ingredients.Agar is a polysaccharide extracted from seaweed. Its coagulation process is associated with the formation of hydrogen bonds. This experiment used agar as the test subject. If a substance added will damage its coagulation, it may also damage the hydrogen bonding. In order to examine whether the destruction of coagulation has any association with toxicity, the raw materials of many kinds of cosmetics were tested in this experiment to observe the correlation between the extension of coagulation time with the IC50 cytotoxicity and the LD50 acute toxicity of products taken orally by mouse. Its accuracy, specificity and sensibility were tested and compared with cosmetics safety testing results. The results showed that compared to a coagulation experiment with LD50 toxicity orally taken by mouse, its accuracy was 73.6%, sensibility 43.7%, specificity 95.4%, false positive 23.6% and false negative 2.6%; compared to the coagulation experiment with B16-F10 cytotoxicity, accuracy was 84.2%, sensibility 60%, specificity 100%, false positive 15.7% and false negative 0%; compared with 3T3 cytotoxicity, accuracy was 89.4%, sensibility 73.3%, specificity 100%, false positive 10.5% and false negative 0%. Compared with the cosmetics safety testing results, coagulation experiment had a lower false negative rate. Based on the above results, the coagulation experiment seems applicable as one of the testing methods for non-animal cosmetics safety testing. It could reduce the use of animals in the preliminary stage of experiments. |
