Genotoxicity of selected pharmaceuticals, their binary mixtures, and varying environmental conditions – study with human adenocarcinoma cancer HT29 cell line

Abstract

Pharmaceutical residues are present in the environment in mixtures and their adverse effects may also result from interactions that occur between compounds. Studies presented in this work focus on genotoxicity of pharmaceuticals from different therapeutic groups in mixtures and in individual solutions impacted with different environmental conditions assessed using comet assay (alkaline approach). Binary mixtures of pharmaceuticals (in different concentration ratios) and in individual solutions impacted with pH change (range from 5.5 to 8.5) or addition of inorganic ions, were incubated with HT29 cells and after 24 h time period cells were tested for the presence of DNA damage. To estimate whether mixtures act more (synergistic) or less (antagonistic) efficiently Concentrations Addition (CA) and Independent Action (IA) approaches were applied followed by a calculation of the Model Deviation Ratio (MDR) to determine deviation from the predicted values. Addition of inorganic ions mainly reduced their genotoxicity. Diclofenac s. was the most susceptible to potassium, fluoride, and bromide ions. Change of the pH of pharmaceutical solutions had significant impact on genotoxicity of diclofenac s. and fluoxetine h. Among mixtures, more commonly observed interactions were synergistic ones, exactly twenty-five cases (ten pairs containing chloramphenicol or oxytetracycline h.) and ten cases of antagonism (four for pairs containing chloramphenicol or fluoxetine h.). The results obtained indicate that interactions between tested compounds occur frequently and can lead to DNA damage. This topic especially concerning in vitro tests using cells is still rare, however, it should not be neglected.

Keywords:

• Drugs residues
• drugs mixtures
• cytotoxicity
• genotoxicity
• MTT
• comet assay

Acknowledgments

Acknowledgments are given to Professor Agnieszka Bartoszek and PhD Eng. Izabela Koss-Mikołajczyk from the Department of Chemistry, Technology, and Biotechnology of Food at Gdańsk University of Technology for sharing the equipment, knowledge and cell line for studies.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

Manuscript was co-funded by the National Science Centre, Poland, based on decision number DEC-2013/09/N/NZ8/03247.