Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Apr 21;9(5):92.
doi: 10.3390/toxics9050092.

Cytotoxicity and Microbicidal Activity of Commonly Used Organic Solvents: A Comparative Study and Application to a Standardized Extract from Vaccinium macrocarpon

Yana Ilieva  1 , Lyudmila Dimitrova  1 , Maya Margaritova Zaharieva  1 , Mila Kaleva  1 , Petko Alov  2 , Ivanka Tsakovska  2 , Tania Pencheva  2 , Ivanka Pencheva-El Tibi  3 , Hristo Najdenski  1 , Ilza Pajeva  2
Affiliations

Cytotoxicity and Microbicidal Activity of Commonly Used Organic Solvents: A Comparative Study and Application to a Standardized Extract from Vaccinium macrocarpon

Yana Ilieva et al. Toxics. .

Abstract

The cytotoxicity and microbicidal capacity of seven organic solvents commonly applied for studying plant extracts and bioactive compounds were systematically investigated based on international standards. Four cell lines of normal (CCL-1, HaCaT) or tumor (A-375, A-431) tissue origin, seven bacterial and one fungal strain were used. The impact of the least toxic solvents in the determination of in vitro cytotoxicity was evaluated using a standardized extract from Vaccinium macrocarpon containing 54.2% v/v proanthocyanidins (CystiCran®). The solvents ethanol, methoxyethanol and polyethylene glycol were the least cytotoxic to all cell lines, with a maximum tolerated concentration (MTC) between 1 and 2% v/v. Ethanol, methanol and polyethylene glycol were mostly suitable for antimicrobial susceptibility testing, with minimum inhibitory concentrations (MICs) ≥ 25% v/v. The MTC values of the solvents dimethyl sulfoxide, dimethoxyethane and dimethylformamide varied from 0.03% to 1.09% v/v. The MICs of dimethyl sulfoxide, methoxyethanol and dimethoxyethane were in the range of 3.125-25% v/v. The cytotoxic effects of CystiCran® on eukaryotic cell lines were directly proportional to the superimposed effect of the solvents used. The results of this study can be useful for selecting the appropriate solvents for in vitro estimation of the cytotoxic and growth inhibitory effects of bioactive molecules in eukaryotic and prokaryotic cells.

Keywords: cell lines; in vitro cytotoxicity; microbicidal effect; organic solvents; proanthocyanidins.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
(a) IC50 and (b) MTC of six solvents expressed as % (v/v) of solvents on a panel of four cell lines after 72 h incubation; (c) numerical values of IC50 and MTC based on three independent assays (n = 3) and used in (a,b), respectively. Legend: IC50—median inhibitory concentration; MTC—maximum tolerated concentration; CI 95%—confidence interval 95%. Asterisks (*) denote that the IC50 or MTC values exceed the maximum tested concentration.
Figure 2
Figure 2
(a) IC50 and (b) MTC of CystiCran® dissolved in four different solvents on a panel of four cell lines after 72 h of incubation. (c) Numerical values of IC50 and MTC based on three independent assays (n = 3) and used in (a,b), respectively. Legend: IC50—median inhibitory concentration; MTC—maximum tolerated concentration; CI 95%—confidence interval 95%. Asterisks (*) denote that the IC50 or MTC values exceed the maximum tested concentration.
See this image and copyright information in PMC

References

    1. ISO 10993-1:2009. Biological Evaluation of Medical Devices—Part 1: Evaluation and Testing within a Risk Management Process. International Organization for Standardization; Geneva, Switzerland: 2017.
    1. ISO 10993-5:2009. Biological Evaluation of Medical Devices—Part 5: Tests for In Vitro Cytotoxicity. International Organization for Standardization; Geneva, Switzerland: 2017.
    1. ISO 20776/1-2006. Clinical Laboratory Testing and In Vitro Diagnostic Test Systems—Susceptibility Testing of Infectious Agents and Evaluation of Performance of Antimicrobial Susceptibility Test Devices—Part 1: Reference Method for Testing the In Vitro Activity of Antimicrobial Agents against Rapidly Growing Aerobic Bacteria Involved in Infectious Diseases. International Organization for Standardization; Geneva, Switzerland: 2006. p. 19. European Committee for Standardization (CEN), Technical Committee CEN/TC 140, Technical Committee ISO/TC 212.
    1. Smith C.G., Lummis W.L., Grady J.E. An improved tissue culture assay. II. Cytotoxicity studies with antibiotics, chemicals, and solvents. Cancer Res. 1959;19:847–852. - PubMed
    1. Holmberg B., Malmfors T. The cytotoxicity of some organic solvents. Environ. Res. 1974;7:183–192. doi: 10.1016/0013-9351(74)90149-2. - DOI

LinkOut - more resources