The dosing determines mutagenicity of hydrophobic compounds in the Ames II assay with metabolic transformation: Passive dosing versus solvent spiking

Smith, Kilian E. C.; Heringa, Minne B.; Uytewaal, Marijan; Mayer, Philipp

doi:10.1016/j.mrgentox.2012.07.006

Cited by 32 publications

(26 citation statements)

References 34 publications

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“…Passive dosing has successfully been applied and tailored toward various standard in vitro assays (Booij et al, 2011;Kramer et al, 2010;Oostingh et al, 2015;Smith et al, 2013a) and has shown to improve the sensitivity of the assays while facilitating a better understanding of the obtained toxicological data.…”

Section: Passive Dosingmentioning

confidence: 99%

Endocrine activity of persistent organic pollutants accumulated in human silicone implants — Dosing assays by partitioning from silicone

Gilbert

Mayer

Pedersen

et al. 2015

Environment International

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Section: Passive Dosingmentioning

confidence: 99%

Endocrine activity of persistent organic pollutants accumulated in human silicone implants — Dosing assays by partitioning from silicone

Gilbert

Mayer

Pedersen

et al. 2015

Environment International

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“…An alternative way to evaluate the Ames MPF results is to use a detection limit based on the binomial formula employed by Smith et al [ 24 ], since Ames test responses are not normally distributed, but follow a binomial distribution. -aminocytidine, 9 -AAC 9 aminoacridine, 2 -AA 2-aminoanthracene a Different concentrations of 2-AA may be used depending on the S9-inducing agent (Aroclor-1254 or phenobarbital/ ß-naphthofl avone), on the different Salmonella strains, and on the actual S9 batch.…”

Section: Preparation Of Strains and Mediamentioning

confidence: 99%

The Ames II and Ames MPF Penta I Assay: A Liquid Microplate Format Modification of the Classic Ames Test

Flückiger-Isler¹,

Kamber²

2014

Genotoxicity and DNA Repair

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This chapter describes a protocol modifi cation of the bacterial reverse mutation test (Ames test). It is based on the same principle as the Ames test but uses a liquid, low-volume microplate version of the fl uctuation method. Clear strengths are the low compound requirement and the increased throughput as compared to the standard format. The liquid system allows for processing several replicates at once with the possibility of using pipetting robots and has an easy colorimetric readout. The Ames II/Ames MPF also uses less S9 and produces less hazardous waste due to the low-volume multiwell format.

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“…For an HOC requiring metabolic activation for its genotoxic activity, passive dosing brings additional benefi ts [ 23 ]. The high solubility of the HOC in the dosing polymer means a large mass of test compound can be introduced into the assay.…”

Section: Control Of Hydrophobic Compound Exposurementioning

confidence: 99%

“…The C Free -based effect concentrations when passive dosing were 3-33 times lower than those based on the nominal spiked concentrations, with the differences increasing for the more hydrophobic compounds due to their enhanced sorption and thus lower availability. The same silicone format was applied in the Ames II assay to determine the concentration-response of benzo(a)pyrene after metabolic activation by S9 [ 23 ]. When spiking, nominal concentrations had to exceed aqueous solubility before any mutagenicity was observed.…”

Section: Control Of Hydrophobic Compound Exposurementioning

confidence: 99%

“…In most cases a simple pre-equilibration step can be incorporated into the in vitro test protocol to ensure equilibrium partitioning prevails. For example, in Smith et al [ 23 ] the benzo(a)pyrene loaded silicone was left to equilibrate overnight with the medium prior to starting the exposure of the bacterial cells in the presence of S9 activation. When passive dosing is used in treatment regimens with metabolic activation, HOC loss kinetics due to the (bio)transformation process require consideration.…”

Section: Step 21: Passive Dosing Phase Depletionmentioning

confidence: 99%

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The Control of Hydrophobic Compound Exposure in In Vitro Tests for Genotoxicity

Smith

2014

Genotoxicity and DNA Repair

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Several of the OECD recommended tests for genotoxicity are in vitro tests, involving the use of open plastic vessels and culture plates, rich culture media, or metabolic activation. These result in volatile, sorptive, and biotransformation losses of the test compound, and thus to poorly defi ned exposure. This has implications for the apparent sensitivity of the assay as well as for establishing a reliable relationship between concentration and response. These issues are particularly relevant for hydrophobic organic compounds (HOCs) which are diffi cult to initially dissolve in aqueous media and particularly susceptible to sorptive losses. Therefore, the in vitro genotoxicity testing of HOCs requires techniques that compensate for losses and lead to defi ned and constant dissolved exposure concentrations. Here, passive dosing can play a useful role. Passive dosing involves a dominating reservoir of polymer-sorbed HOC acting as a partitioning source to the test medium, maintaining defi ned and constant dissolved concentrations. This chapter introduces passive dosing for the control of HOC dissolved concentrations during in vitro genotoxicity tests, covering considerations for selecting a suitable passive dosing polymer and its dimensioning, as well as procedures for cleaning and loading the dosing polymer with the test HOC(s).

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The dosing determines mutagenicity of hydrophobic compounds in the Ames II assay with metabolic transformation: Passive dosing versus solvent spiking

Cited by 32 publications

References 34 publications

Endocrine activity of persistent organic pollutants accumulated in human silicone implants — Dosing assays by partitioning from silicone

Endocrine activity of persistent organic pollutants accumulated in human silicone implants — Dosing assays by partitioning from silicone

The Ames II and Ames MPF Penta I Assay: A Liquid Microplate Format Modification of the Classic Ames Test

The Control of Hydrophobic Compound Exposure in In Vitro Tests for Genotoxicity

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