Validation of the red blood cell test system as in vitro assay for the rapid screening of irritation potential of surfactants.

Molecular toxicology Pub Date : 1987-09-01
W J Pape, U Pfannenbecker, U Hoppe
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Abstract

An in vitro red blood cell assay (RBC assay) is presented that allows one to estimate irritation potentials of tensides and detergents. The estimation is based on the differentiation between cell membrane lysis and cell protein denaturation. Both effects are measured photometrically by use of the inherent native dye oxyhemoglobin (HbO2). Besides hemolysis (H50) as a test parameter, the denaturation index DI is introduced, which is defined to be equal to 100% at a concentration of 30 mol sodium dodecylsulfate (SDS) per mole HbO2 as internal standard. The HbO2 release (H50), its denaturation (DI), and the ratio of both parameters (L/D ratio) are used to characterize the in vitro effects of surfactants. All data, including the L/D ratio are compared with in vivo data on eye irritancies, evaluated according to OECD Guideline #405 for testing chemicals, and with other published results from in vivo experiments. The good correlation of the L/D ratio of a broad range of 100 marketable shampoos with their corresponding Draize data (r = .806, p less than .0001) allows one to predict eye irritation potentials from another 20 commercially available shampoos in a blind trial with highly significant rank correlations to their in vivo irritancies (rs = .911, p less than .0001). Nearly similar good correlations were obtained by comparing ranks of in vivo and in vitro data of 16 anionic surfactants. All correlations found were significant (rs greater than .80 and p less than .0001). The RBC assay is an inexpensive, rapid, irritancy screening test that provides reliable results with good reproducibility. The test helps to reduce or even avoid animal testing in this application.

验证红血球测试系统作为快速筛选表面活性剂刺激潜能的体外试验。
体外红细胞测定(RBC测定)提出,允许一个估计张力和洗涤剂的刺激电位。估计是基于细胞膜裂解和细胞蛋白变性之间的区别。这两种效应都是用固有的天然染料氧合血红蛋白(HbO2)光度法测量的。除了溶血(H50)作为测试参数外,还引入了变性指数DI,定义为在每摩尔HbO2浓度为30 mol十二烷基硫酸钠(SDS)为内标时,DI等于100%。用HbO2的释放量(H50)、变性率(DI)和两者的比值(L/D比)表征表面活性剂的体外作用。将所有数据(包括L/D比率)与根据经合组织第405号化学品测试指南评估的体内眼睛刺激数据进行比较,并与其他已发表的体内实验结果进行比较。广泛的100种市场上销售的洗发水的L/D比率与其相应的Draize数据(r = 0.806, p小于0.0001)的良好相关性允许人们在盲试验中预测另外20种市售洗发水的眼睛刺激电位,其体内刺激具有高度显著的等级相关性(rs = 0.911, p小于0.0001)。通过比较16种阴离子表面活性剂的体内和体外数据,得到了几乎相似的良好相关性。所有发现的相关性均显著(rs大于0.80,p小于0.0001)。红细胞分析是一种廉价、快速、刺激筛选试验,可提供可靠的结果和良好的重复性。该测试有助于减少甚至避免在此应用程序中进行动物测试。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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