Research of cytotoxic activity of drinking water using methods in vitro

  • M. R. Vergolyas International Academy of Ecology and Medizine, Bogdan Khmelnitsky Melitopol State Pedagogical University
  • D. V. Kovalenko Bogdan Khmelnitsky Melitopol State Pedagogical University
  • I. O. Khaliman Bogdan Khmelnitsky Melitopol State Pedagogical University
  • M. V. Vikhliaieva Bogdan Khmelnitsky Melitopol State Pedagogical University
  • K. O. Molozhan Bogdan Khmelnitsky Melitopol State Pedagogical University
  • О. О. Verholias Bogdan Khmelnitsky Melitopol State Pedagogical University


Aim. The aim of this work is to assess the quality of drinking water of various origins by its cytotoxic effect on human and animal cell cultures in in vitro experiments. Methods. The cytotoxic effect of control water obtained in accordance with the requirements of DSTU 4174: 2003, tap water, pump room and packaged Evian water was studied. The studies were carried out on NEC-293 cells (human kidney cells) and L929 (mouse fibroblasts) cells from the Institute of Microbiology and Virology named after V.I. D.K. Zabolotny National Academy of Sciences of Ukraine. Results. Studies in the MTT test of cytotoxicity of these water samples showed that tap water and bottled water exhibited the greatest toxic activity against human kidney cells of the NEC-293 line (the number of viable cells under their influence was 61.84% and 79.06%, respectively). Under the influence of water from the pump room - 84.55%, the control water showed the least influence - 93.13%. In the test with sulforhodamine B, the number of living cells under the influence of control water was 100.00%, tap water 80.15%, water from pump rooms 97.11%, Evian 84.70%. Conclusions. The greatest cytotoxic effect according to the MTT test and the test with sulforhodamine B on human kidney cells of the HEK-293 line and fibroblasts of the L929 mouse was exhibited by tap and bottled water. The least influence on the viability of the cells was exerted by the water from the pump room and the control. The negative effect of water on cell viability was manifested in the dysfunction of mitochondria and protein synthesis.

Keywords: cytotoxicity, cell culture, drinking water.


Serdyuk A.M., Polka N.S., Makhnyuk V.M. Modern problems of hygiene of planning and building of settlements (normative-legal regulation). Kyiv: Medinform, 2014. 174 p. [in Ukraine]

Prokopov V.O. Drinking water of Ukraine: medical-ecological and sanitary-hygienic aspects. Кyiv: «Medicine», 2016. 400 p. [in Ukrainian].

Vergolyas M. Сytogenetic evaluation of the drinking water toxicity. EUREKA: Life Sciences. 2016. Vol. 1. Р. 47–54. doi: 10.21303/2504-5695.2016.00054

Trachtenberg I.M. Alternative methods and test systems. Kyiv: VD "Avicenna", 2008. 268 p. [in Ukrainian]

Clemedson C., Kolman A., Forsby A. The Integrated Acute Systemic Toxicity Project (ACuteTox) for the optimisation and validation of alternative in vitro tests. Altern Lab Anim. 2007. Vol. 35. doi: 10.1177/026119290703500102.

Vergolyas M.R., Trachtenberg I.M., Dmitrukha I.M. Estimation of cytotoxic activity of water from different sources of water supply. Environment and Health. 2016. Vol. 4 (80). P. 19–22 [in Ukrainian].

Combes R.D. The use of human cells in biomedical research and testing. Altern. Lab. Anim. 2004. Vol. 32. P. 43–49.

Cavas T. The evaluation of toxicity and mutagenicity of various drinking waters in the human blood lymphocytes (HULYs) in vitro. Food and Chemical Toxicology. 2008. Vol. 46 (7). P. 247–254.

Trakhtenberg I.M., Ulberg Z.R., Chekman I.S., Dmitrukha N.M. Methodical recommendations. Safety assessment of medicinal nanopreparations. Kyiv, 2011. 108 p. [in Ukrainian].

Lee S.K., Kim H.J., Chi S.G., Jang J.Y. Bioflor suppresses expression of Interleukin-8 in HT-29 cell. Intestinal Res. 2004. Vol. 2 (2). P. 96–101.

Atlas R. M. Handbook of Microbiological Media. 4th edition. London New York: Boca Raton: CRC Press; 2010.

Jozefczuk J., Drews K., Adjaye J. Preparation of mouse embryonic fibroblast cells suitable for culturing human embryonic and induced pluripotent stem cells. J Vis Exp. 2012. Vol. 64. P. 3854. doi: 10.3791/3854.

Vergolyas M. Research of cytotoxic activity of water from different water supply sources. Adv Tissue Eng Regen Med Open Access. 2019. Vol. 5 (3). Р. 92‒96. doi: 10.15406/atroa.2019.05.00105.

Yakubchak О.М., Zagrebelny V.O., Adamenko L.V. Regarding the toxicity of chlorinated tap water. Scientific Bulletin of the National University of Life and Environmental Sciences of Ukraine. 2013. Vol. 188 (3). P. 182–185.