改良光鸡胚试验评估化妆品用超细二氧化钛的光毒性

doi:10.3969/j.issn.1001-1803.2021.02.006

摘要/Abstract

摘要：

通过对德国人Neumann的光鸡胚试验进行了改良,并对其评估化妆品用超细二氧化钛的光毒性方面进行了初步应用。实验采用2×2阶测试设计,将受试物以无毒浓度作用于培育9 d的鸡胚绒毛尿囊膜（CAM）,继而暴露于5 J/cm ²的UVA紫外区,去除受试物后再孵育24 h,观察CAM血管反应并计算光刺激指数,区分光毒性效力。通过对10种不同粒径、晶型、表面涂层的超细二氧化钛原料的应用研究发现,该改良光鸡胚试验可作为一种对超细无机防晒原料进行光安全性筛选的简便快捷、高灵敏度的体外替代方法。

关键词: 鸡胚绒毛尿囊膜, 光毒性, 动物替代试验, 超细二氧化钛

Abstract:

A modified Neumann’s photo hen’s egg test was developed and applied in the phototoxicity assessment of ultrafine titanium dioxide ingredients used in cosmetics. The test substance was directly applied onto 9-day chicken embryo chorioallantoic membrane （CAM） at non-toxic dosage. After exposure to 5 J/cm ² UVA, the test substance was removed from CAM and eggs were incubated for 24 hours. The vascular responses of CAM and the embryo lethality were observed and a phototoxic index was calculated to quantify the phototoxic potential. Ten ultrafine titanium dioxide samples differing in particle size, crystal form and surface coating were investigated, suggesting that this improved photo-CAM assay could be used as a simple, rapid and sensitive in vitro method in the photosafety screening of ultrafine inorganic sunscreen materials.

Key words: chicken embryo chorioallantoic membrane, phototoxicity, alternative to animal testing, ultrafine titanium dioxide

中图分类号:

TQ658

刘磊,孔雪,唐颖. 改良光鸡胚试验评估化妆品用超细二氧化钛的光毒性[J]. 日用化学工业, 2021, 51(2): 115-120.

LIU Lei,KONG Xue,TANG Ying. Application of a modified photo hen’s egg test to assess the phototoxicity of ultrafine titanium dioxide particles used in cosmetics[J]. China Surfactant Detergent & Cosmetics, 2021, 51(2): 115-120.

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光照	受试物
光照	有	无
有	n=12	n=12
无	n=12	n=12

得分	血管反应
0	CAM无可见变化
1	仅轻微可见的出血、凝血以及膜变色
2	部分可见的出血、凝血以及膜变色
3	几乎完全覆盖的出血、凝血以及膜变色

样品编号	表面性质	表面涂层	晶型	粒径/nm	^*3T3预测结果
TiR10OC	亲油	硅石/聚二甲基硅氧烷	金红石	10~30	N/A
TiR10HC	亲水	硅石	金红石	10~30	-
TiR30OC	亲油	硅油	金红石	30~50	N/A
TiAR30HC	亲水	二氧化硅/氧化铝等	金红石/锐钛矿	30~50	+
TiAR30OC	亲油	月硅酸/硬脂酸等	金红石/锐钛矿	30~50	N/A
P25	亲水		金红石/锐钛矿	10~30	?
TiAR30H	亲水		金红石/锐钛矿	30~50	-
TiAR30O	亲油		金红石/锐钛矿	30~50	N/A
TiA10O	亲水		锐钛矿	10~30	-
TiAR100H	亲水		金红石/锐钛矿	≥100	-

样品	质量浓度/ （μg·mL^-1）	光致死率/ %	1~3级光致效应			光毒性指数
样品	质量浓度/ （μg·mL^-1）	光致死率/ %	R₁/%	R₂/%	R₃/%	光毒性指数
氯丙嗪	0.01	33.33	4.17	37.5	50	3.63
8-甲氧基补骨脂	2.5	0	8.33	16.67	75	2.75
L-抗坏血酸	1 000	8.33	72.73	9.09	18.18	1.79
茶多酚	10 000	8.33	0	8.33	8.33	0.75
L-组氨酸	3	0	8.33	0	0	0.17
TiAR30HC	100	33.33	8.33	50.00	25.00	3.17
TiAR30OC	100	16.67	33.33	33.33	33.33	2.67
TiA10O	100	25.00	58.33	11.11	22.22	2.47
TiR10OC	100	16.67	22.22	19.44	33.33	2.28
P25	100	25.00	25.56	33.33	11.11	2.26
TiAR30O	100	16.67	37.50	33.33	16.67	2.21
TiAR30H	100	16.67	26.26	44.44	11.11	2.15
TiR30OC	100	16.67	31.67	40.00	11.67	2.13
TiR10HC	100	16.67	30.00	27.50	12.50	1.89
TiAR100H	100	8.33	36.67	2.22	11.11	1.08