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Validation Cytotoxicity Assay for Lipophilic Substances.
Natalia Mencacci Esteves-Pedro1, Kenji Sugibayashi2, Elissa A Ostrosky3
1Departamento de Farmacia, Faculdade de Ciencias Farmaceuticas, Universidade de Sao Paulo, Sao Paulo, Brazil.
Current Topics in Medicinal Chemistry
|April 12, 2018
Summary
Dispersing lipophilic essential oils in cytotoxicity assays is challenging. Polysorbate 20, guided by Hydrophile-Lipophile Balance theory, effectively solubilized these oils, improving assay reliability.
Area of Science:
- Pharmacology
- Toxicology
- Analytical Chemistry
Background:
- Lipophilic substances (log Kow ≥ 5) pose dispersion challenges in validated cytotoxicity assays, potentially causing false negatives.
- Essential oils from Minthostachys setosa, Pimenta pseudocaryophyllus, and Drimys brasiliensis are lipophilic and require effective dispersion methods.
Purpose of the Study:
- To elucidate challenges in validated cytotoxicity testing of lipophilic essential oils.
- To compare the efficacy of Neutral Red (NR) and MTS assays for cell viability detection.
- To identify optimal solubilizing agents for lipophilic substances using Hydrophile-Lipophile Balance (HLB) theory.
Main Methods:
- Evaluation of essential oil dispersion and cytotoxicity using the Hydrophile-Lipophile Balance (HLB) technique.
- Comparison of Neutral Red (NR) and MTS vital dyes for cell viability assessment via TOST equivalence test.
- Adherence to OECD / GD 129 guidelines for validated cytotoxicity assays.
Main Results:
- Demonstrated alternative methods for dispersing lipophilic substances in cytotoxicity assays.
- Identified polysorbate 20 as the most effective solubilizing agent for the studied essential oils in D10 medium based on HLB theory.
- Confirmed the potential for improved accuracy in cytotoxicity testing of lipophilic compounds.
Conclusions:
- Effective dispersion of lipophilic substances is crucial for accurate cytotoxicity assay results.
- Hydrophile-Lipophile Balance (HLB) theory provides a robust framework for selecting appropriate solubilizing agents.
- Polysorbate 20 is a suitable agent for enhancing the solubility of Minthostachys setosa, Pimenta pseudocaryophyllus, and Drimys brasiliensis essential oils in cytotoxicity assays.
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