Comparison of Two Methods for Preparing Mitochondria from Tomato Leaves to Study the Ascorbate-Glutathione Cycle Activity

Mitochondria were purified from tomato leaves by a differential centrifugation method and a Percoll density-gradient method. A comparative study on the ascorbate-glutathione cycle activity revealed that there was no difference in the specific activity of the antioxidative enzymes and ascorbate and glutathione concentrations between mitochondria prepared by these two methods. However, the contents of dehydroascorbic acid and oxidised glutathione in Percoll-separated organelles were 260 and 210 %, respectively, of those found in mitochondria purified by the differential centrifugation method.