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Joint International Conference on Long-term Experiments ...

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attempted accommodati<strong>on</strong> to the diet, it was decided to carry out the experiment with twothirds<br />

of the recommended ethanol level (4.3 % of the diet, 24% of total calories). NDMA<br />

was included in both ethanol and c<strong>on</strong>trol diets at 10 ppm, and the mice were pair-fed for 2<br />

weeks so that mice were given NDMA-c<strong>on</strong>trol diet in amounts equal to that c<strong>on</strong>sumed by<br />

the mice given NDMA –ethanol diet.<br />

Estimati<strong>on</strong> of Tissue C<strong>on</strong>tent of NDMA. The amount of NDMA in samples of blood and<br />

other organs (pooled from 5-10 mice) was estimated by methylene chloride extracti<strong>on</strong>thermal<br />

energy analysis as described previously. The limit of detecti<strong>on</strong> of NDMA by this<br />

procedure was 0.5-1 ppb, depending <strong>on</strong> the amount of starting material. Water blanks and<br />

tissues from c<strong>on</strong>trol mice were repeatedly assayed and did not c<strong>on</strong>tain measurable levels of<br />

NDMA (

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