An artificial aqueous humor as a standard matrix to assess drug concentration in the anterior chamber by high performance liquid chromatography methods.

Abstract

The aim of the study was to recreate in-vitro artificial aqueous humor with the same physico-chemical properties of human aqueous humor to be used as a standard matrix in chromatography to assess drug concentration in the anterior and posterior chamber of the human eye. The artificial aqueous humor was prepared according to the human aqueous humor chemical compositions reported in the literature. The artificial matrix was then analysed via the HPLC-UV method and compared with aqueous humor from 15 patients who underwent cataract surgery. Known concentrations of widely-used ophthalmological drugs were added to the artificial aqueous humor in order to assess whether it can be used to explore ocular disposition towards topically or systemically administered drugs. No significant differences were found between the two examined aqueous humor types. There were no significant qualitative differences between examined fluids in terms of presence of ophthalmological drugs. The composition of artificial, in-vitro recreated aqueous humor was similar to that of the human kind. The absence of significant differences in the analysis of tested drugs both in the artificial and in human aqueous humor indicates that artificial aqueous humor may be used to generate a matrix-based standard curve for analytical method validation.