Rapid sample preparation for determination of iron in tissue by closed-vessel digestion and microwave energy.

We developed a rapid acid-digestion method for preparing tissue samples for iron determination. Specimens were digested in nitric acid and hydrogen peroxide under high temperature and pressure in closed Teflon vessels, with microwave energy. Analysis for iron in 25- to 250-mg portions of digested bovine liver powder (National Bureau of Standards Certified Reference Material no. 1577a) showed excellent linearity ([predicted] = 1.007[actual] - 0.166 micrograms per sample) and analytical recovery (98%). Precision (CV) was 5.4% when iron content was 10 micrograms per sample. Assaying split samples of mouse tissues, we found a close correlation between iron concentrations obtained with closed vs open vessels ([closed] = 0.878[open] + 68 micrograms/g, r = 0.994, range 400-4600 micrograms/g dry weight). In contrast to time-consuming conventional procedures for tissue dissolution, closed-vessel digestion with microwave energy dramatically shortens time for tissue preparation, minimizes use of caustic acid, reduces risk of sample loss or contamination, and yields accurate and reproducible results.