Improvement of the Bovine Corneal Opacity and Permeability (BCOP) assay as an in vitro alternative to the Draize rabbit eye irritation test

Summary The Draize rabbit eye irritation test has several disadvantages and inadequacies when used as an indicator for potential irritancy of detergent-based commercial products. As a possible in vitro screen, it was decided to use mouse LS cells cultured in suspension, taking 50% cell death (CD50) after exposure to the product for 4h as the endpoint. This figure for 11 formulations was compared with eye irritation data ranked using an arbitrary classification of mild, moderate and severe. All samples with a CD50 less than 0.5mg/ml were severe eye irritants, while most of those with a CD50 greater than 1.0mg/ml were mild irritants. Samples between these two cytotoxicity levels were, in general, moderately irritant to the rabbit eye. It would appear that this in vitro test is a possible screen for the irritancy of detergent-based products.

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Studies of an in vitro alternative method to the draize rabbit eye irritation test. Comparison of hemolysis of erythrocytes and neutral red uptake in culture cell.

Journal of Society of Cosmetic Chemists of Japan ◽

10.5107/sccj.23.272 ◽

1990 ◽

Vol 23 (4) ◽

pp. 272-279 ◽

Cited By ~ 10

Author(s):

Yuuko Okamoto ◽

Noriko Kanzaki ◽

Nobuyuki Tanaka

Keyword(s):

Neutral Red ◽

Culture Cell ◽

Eye Irritation ◽

Rabbit Eye ◽

Alternative Method ◽

Irritation Test ◽

Neutral Red Uptake

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In Vitro Effectiveness of Neem Oil (Azadirachta Indica A. Juss) Shampoo as Anti Head Lice (Pediculus humanus capitis)

Pharmacology and Clinical Pharmacy Research ◽

10.15416/pcpr.v4i3.25780 ◽

2019 ◽

Vol 4 (3) ◽

pp. 76

Author(s):

Norisca A. Putriana ◽

Azizul Hakim ◽

Patihul Husni ◽

Taofik Rusdiana

Keyword(s):

Azadirachta Indica ◽

Corneal Opacity ◽

Head Lice ◽

Neem Oil ◽

Pediculus Humanus Capitis ◽

Eye Irritation ◽

Pediculus Humanus ◽

Study Results ◽

Irritation Test

Head lice (Pediculus humanus capitis), a parasite causing hair problem, often occurs in society.The study aimed to evaluate in vitroeffectiveness of shampoo formulation containing neem oil as an anti head lice. An experimental method was used to examine the formulation of shampoo, irritation test, testing the effectiveness of shampoo against head lice (Pediculicidal activity), and testing the effectiveness of shampoo against nits (Ovicidal activity). The effectiveness test employed an in vitromethod where neem oil shampoo were tested directly towards head lices and nits isolated from humans, then the mortality was observed. To recognize the safety of the shampoo, skin and eye irritation tests were carried out by applying 0.5 ml on the skin and dripping 0.1 ml shampoo into New Zealand white rabbit’s eyes.Data were analyzed using Kruskal Wallis and Mann Whitney. The study results showed that the primary irritation index for 5%, 10%, 15%, and 20% neem oil shampoo were 0; 0111; 0.222; and 0.222, respectively. All of the shampoos were very mild irritation response category. Eye irritation tests showed that only the 20% neem oil shampoo gave a mild irritation response with degree of corneal opacity ≥ 1 and recovered after 7 days of observation. The 5%, 10%, 15%, and 20% neem oil shampoo were effective as an anti head lice. The 15% and 20% neem oil shampoo had anti head lice effectiveness activity as same as 1% permethrin lotion. In conclusion, the 15% neem oil shampoo was chosen as the best shampoo, its high anti head lice effect and light irritation response. Keywords: Head lice, neem oil (Azadirachta indica A. Juss), shampoo

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An evaluation of five potential alternatives in vitro to the rabbit eye irritation test in vivo

Toxicology in Vitro ◽

10.1016/0887-2333(92)90017-l ◽

1992 ◽

Vol 6 (4) ◽

pp. 275-284 ◽

Cited By ~ 49

Author(s):

D.M. Bagley ◽

L.H. Bruner ◽

O. de Silva ◽

M. Cottin ◽

K.A.F. O'Brien ◽

...

Keyword(s):

Eye Irritation ◽

Rabbit Eye ◽

Irritation Test

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Using In Vitro Prediction Models Instead of the Rabbit Eye Irritation Test to Classify and Label New Chemicals: A Post Hoc Data Analysis of the International EC/HO Validation Study

Alternatives to Laboratory Animals ◽

10.1177/026119290303100105 ◽

2003 ◽

Vol 31 (1) ◽

pp. 31-46 ◽

Cited By ~ 8

Author(s):

Ferdinand Moldenhauer

Keyword(s):

Validation Study ◽

Random Data ◽

Data Set ◽

Eye Irritation ◽

Mean Values ◽

Rabbit Eye ◽

Irritation Test ◽

The Mean ◽

British Home

The international validation study on alternative methods to replace the Draize rabbit eye irritation test, funded by the European Commission (EC) and the British Home Office (HO), took place during 1992–1994, and the results were published in 1995. The results of this EC/HO study are analysed by employing discriminant analysis, taking into account the classification of the in vivo data into eye irritation classes A (risk of serious damage to eyes), B (irritating to eyes) and NI (non-irritant). A data set for 59 test items was analysed, together with three subsets: surfactants, water-soluble chemicals, and water-insoluble chemicals. The new statistical methods of feature selection and estimation of the discriminant function's classification error were used. Normal distributed random numbers were added to the mean values of each in vitro endpoint, depending on the observed standard deviations. Thereafter, the reclassification error of the random observations was estimated by applying the fixed function of the mean values. Moreover, the leaving-one-out cross-classification method was applied to this random data set. Subsequently, random data were generated r times (for example, r = 1000) for a feature combination. Eighteen features were investigated in nine in vitro test systems to predict the effects of a chemical in the rabbit eye. 72.5% of the chemicals in the undivided sample were correctly classified when applying the in vitro endpoints lgNRU of the neutral red uptake test and lgBCOPo5 of the bovine opacity and permeability test. The accuracy increased to 80.9% when six in vitro features were used, and the sample was subdivided. The subset of surfactants was correctly classified in more than 90% of cases, which is an excellent performance.

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Application of the EYTEX™ System to the Evaluation of Cosmetic Products and their Ingredients

Alternatives to Laboratory Animals ◽

10.1177/026119299202000120 ◽

1992 ◽

Vol 20 (1) ◽

pp. 146-163

Author(s):

Francis H. Kruszewski ◽

Laura H. Hearn ◽

Kyle T. Smith ◽

Janice J. Teal ◽

Virginia C. Gordon ◽

...

Keyword(s):

Double Blind ◽

Eye Irritation ◽

Ocular Irritation ◽

Rabbit Eye ◽

Wide Range ◽

High Concentrations ◽

Alkaline Membrane ◽

Positive Agreement

465 cosmetic product formulations and raw ingredients were evaluated with the EYTEX™ system to determine the potential of this in vitro alternative for identifying eye irritation potential. The EYTEX™ system is a non-animal, biochemical procedure developed by Ropak Laboratories, Irvine, CA, that was designed to approximate the Draize rabbit eye irritation assay for the evaluation of ocular irritation. Avon Products Inc. provided all the test samples, which included over 30 different product types and represented a wide range of eye irritancy. All the EYTEX™ protocols available at the time of this study were used. Samples were evaluated double-blind with both the membrane partition assay (MPA) and the rapid membrane assay (RMA). When appropriate, the standard assay (STD) and the alkaline membrane assay (AMA) were used, as well as specific, documented protocol modifications. EYTEX™ results were correlated with rabbit eye irritation data which was obtained from the historical records of Avon Products Inc. A positive agreement of EYTEX™ results with the in vivo assay was demonstrated by an overall concordance of 80%. The assay error was 20%, of which 18% was due to an overestimation of sample irritancy (false positives) and 2% was attributed to underestimation (false negatives). Overestimation error in this study was due in part to the inability of the protocols to accurately classify test samples with very low irritation potential. Underestimation of sample irritancy was generally associated with ethoxylated materials and high concentrations of specific types of surfactants. 100% sensitivity and 85% predictability were described by the data, indicating the efficiency of EYTEX™ in identifying known irritants. A specificity rate of 39% showed the EYTEX™ assay to be weak in discerning non-irritants. However, the EYTEX™ protocols used in this study were not designed to identify non-irritants. A compatibility rate of 99% proved the effectiveness of the EYTEX™ assay in accommodating a diversity of product types. The EYTEX™ system protocols, when used appropriately, can provide a conservative means of assessing the irritant potential of most cosmetic formulations and their ingredients.

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