Student Research: Denise Bender

, , 1991
Faculty Advisor: David A Kalman

Detection and Analysis of Mutagenicity in Complex Mixtures Using the Combined Ames/TLC Assay


Abstract

The previous Ames/TLC technique is limited in its use for assessing mutagenic potency of samples by the following:

1. Inadequate methods in quantifying the dose-response of the separated compounds.
2. Inadequate separation of polar compounds.
3. Lack of detection of non-polar compounds.

The objectives of this project were to assess modifications of the present method for their ability to overome the obstacles discussed and thereby provide quantitative results. Some of these modifications include:

1. Applying known amounts of mutagenic compounds in etched out areas on thin-layer chromatography plates in the standard assay in order to establish mutagenic zones of response.
2. Selecting stationary and mobile phase(s) suitable for the separation of single compounds and mixtures.
3. Enhancing the sensitivity of the standard plate incorporation method in order to detect non-polar compounds. Modifications include varying the depth of the agar, enhancing the solubility of the test compounds in the agar, and varying the sequence of the layers in the assay.

A second aim of this study was to determine if these modifications enhance the sensitivity of the standard method; and to then relate the quantitative results from the dose-response of the known mutagenic compounds tested singly and in mixtures resolved by TLC.

Taken from the beginning of thesis.