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Histamine determination in tears as an index of safety in use of cosmetic products
Authors:A. BETTERO  M. R. ANGI  F. GALIANO  C. A. BENASSI
Affiliation:Department of Pharmaceutical Sciences, University of Padova, Italy;Department of Ophthalmology, University of Padova, Italy
Abstract:To assess ocular irritancy caused by chemical and cosmetic products a reliable method based on evaluation of histamine (Hm) in tears is presented. Hm is measured at picomole levels by HPLC and fluorimetric detection after fluorescamine-HM derivatization. In order to avoid any uncontrolled irritation and stimulation of the conjunctiva during sample collection, a procedure of conjunctiva lavage was developed. A balanced salt solution (50 μl) containing a known amount of Hm-fluorophore as reference standard is instilled in the conjunctiva fornix. After a few seconds 20 μl of tear fluid is collected: 10 μl are immediately analysed and 10 μl after derivatization reaction. In this way it is possible to evaluate tear dilution and to assess Hm content in less than 10 minutes.
In a group of 20 normal subjects Hm has been determined in comparison with that of two volunteers after topical application of 50 μ of 0.2% and 0.4% sodium lauryl sulphate solution. A contact of 30 seconds of the cosmetic ingredient caused an immediate dose-dependent Hm release through a direct cytotoxic damage of cell membranes due to the surfactant action.
Évaluation de l'irritation occulaire due aux cosmétiques par la détermination de l'histamine lacrimale
Keywords:ocular irritancy    fluorescamine-histamine derivatization    fluorophore reference standard    conjunctival lavage fluid    histamine-release
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