基于in vitro和in silico技术的化学品眼刺激性评价替代方法研究

doi:10.3969/j.issn.1001-1803.2022.09.015

Abstract

Abstract:

The aim of the study was to establish the alternative method for preliminary assessing eye irritation of chemicals using Fluorescein Leakage test （FL test） and Quantitative Structure Activity Relationship （QSAR）, to detect and classify the ocular irritation of the recommended substances, and to explore the application value of different alternative methods. Using Toxtree models to predict eye irritation and irritation potency of 8 recommended substances for demonstrating technical proficiency. The test substances were prepared into series of concentrations and exposed with the Madin-Darby canine kidney cell （MDCK） for 1 min to test the percentage of fluorescein leakage （FL/%）. The results were categorized according to the classification criteria for assessing eye irritation. Then, the sensitivity, specificity, and accuracy of FL test and Toxtree models were evaluated. Furthermore, the consistency of FL test or Toxtree model was compared with UN-GHS （United Nationas Globally Harmonized System of Classification and Labelling of Chemicals） classification. When FL test is used to distinguish if the test substance belongs to severe irritation/corrosion （GHS Cat. 1）, the irritation potency of all test substances is concordant with that in GHS classification （Kappa=1, P=0.005）. When FL test is used to distinguish if the test substance is severe irritation/corrosion （GHS Cat. 1）, moderate irritation （GHS Cat. 2A） or mild/no irritation （GHS Cat. 2B/No Cat.） substances, the irritation potency of all test substances is concordant with GHS classification in the eye irritant potency （Kappa=0.579, P=0.035）. The results predicted by Toxtree software demonstrate that it is more suitable to firstly estimate the possible substances with mild/no ocular irritation. The FL test is an effective and valuable alternative method for evaluating eye irritation, and it is more applicable to identify chemicals with severe irritation/corrosion or to be the study firstly carried out using the “Top-Down approach” of the integrated decision-tree testing strategy for eye irritation. The results predicted by Toxtree software could be reference data for the integrated assessment on eye irritation.

Key words: eye irritation, alternative method, in vitro, quantitative structure-activity relationship, fluorescein leakage test, Toxtree model

CLC Number:

TQ658

Lin Ni,Luo Feiya,Cao Chunran,Hu Yuchi. Study on the alternative method for assessing eye irritation of chemicals using in vitro and in silico approaches[J].China Surfactant Detergent & Cosmetics, 2022, 52(9): 1016-1022.

Figures/Tables 5

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References 19

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眼刺激程度分级	UN GHS C&L ^[11]	MMAS ^[12]	FL₂₀ ^[7]/（mg/mL）	Toxtree ^[13]
无刺激性	No Cat.	<2.5	>750	Nonirritant/not corrosive
轻刺激性	Cat. 2B	2.5~25.0
中度刺激性	Cat. 2A	25.0~50.0	100~750	Moderate reversible irritation to the eye
严重刺激性/腐蚀性	Cat. 1	>50.0	≤100	Serious lesions to the eye /corrosion

序号	受试物	CAS号	化学类别	理化性质
序号	受试物	CAS号	化学类别	物理状态	分子量	pK_a	熔点	c lg P^*
1	苯扎氯铵（5%）	8001-54-5	季铵盐,阳离子表面活性剂	液体	340	N/A	N/A	2.126
2	盐酸异丙嗪	58-33-3	胺/脒,杂环,有机硫化合物	固体	321	9.287	228.5	4.915
3	氢氧化钠（10%）	1310-73-2	碱	液体	40	N/A	323	0.467
4	十二烷基硫酸钠（15%）	151-21-3	羧酸盐	液体	288	N/A	205.5	4.106
5	4-醛基苯甲酸	619-66-9	羧酸,醛	固体	150	4.089	256	1.33
6	2-甲基乙酰乙酸乙酯	609-14-3	酮,酯	液体	144	N/A	187	0.924
7	月桂醇聚氧乙烯醚 Brij-35（30%）	9002-92-0	脂肪醇,醚,非离子型表面活性剂	液体	1 199.5	14.436	40~45	4.28
8	甘油	56-81-5	醇	液体	92.1	17.965	18	-1.88

序号	UN-GHS眼刺激性类别	Draize test		FL test		Toxtree
序号	UN-GHS眼刺激性类别	MMAS	分级	FL₂₀/ （mg/mL）	分级	结果	分级	警示结构
1	1	108	严重刺激性/腐蚀性	6.33	严重刺激性/腐蚀性	中度可逆性眼刺激 Moderate reversable irritation to the eye	中度刺激性	芳香盐铵 Aromatic ammonium salts
2	1	71.7	严重刺激性/腐蚀性	12.43	严重刺激性/腐蚀性	非皮肤腐蚀性 NOT skin corrosion R34 or R35	N/A
3	1	108	严重刺激性/腐蚀性	9.73	严重刺激性/腐蚀性	非皮肤腐蚀性 NOT skin corrosion R34 or R35	N/A
4	1	59.2	严重刺激性/腐蚀性	8.58	严重刺激性/腐蚀性	非皮肤腐蚀性 NOT skin corrosion R34 or R35	N/A
5	2A	50.0	中度刺激性	>750	轻刺激性/无刺激性	非刺激性Nonirritant	无刺激性
6	2B	18	轻刺激性	>750	轻刺激性/无刺激性	非刺激性Nonirritant	无刺激性
7	2B	16	轻刺激性	133.82	中度刺激性	未知分类Unknown	N/A
8	No Cat.	1.7	无刺激性	>750	轻刺激性/无刺激性	未知分类Unknown	N/A

眼刺激程度分级/结果		GHS结果		灵敏度/%	特异度/%	准确度/%
眼刺激程度分级/结果		阳性（4）	阴性（4）	灵敏度/%	特异度/%	准确度/%
FL test	阳性	4	0	100	100	100
FL test	阴性	0	4	100	100	100
Toxtree	阳性	0	0	0	50	25
Toxtree	阴性	1	2	0	50	25

眼刺激程度分级/结果		GHS分级			准确度/%
眼刺激程度分级/结果		严重刺激性/腐蚀性（4）	中度刺激性（1）	无刺激性/轻刺激（3）	准确度/%
FL test	严重刺激性/腐蚀性	4	0	0	75
	中度刺激性	0	0	1
	无刺激性/轻刺激性	0	1	2
Toxtree	严重刺激性/腐蚀性	0	0	0	12.5
	中度刺激性	1	0	0
	无刺激性/轻刺激性	0	1	1

Study on the alternative method for assessing eye irritation of chemicals using in vitro and in silico approaches

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