The use of high-throughput data to study the changing behavior of biological pathways has focused mainly on examining the changes in the means of pathway genes. In this paper, we propose instead to test for changes in the co-regulated and unregulated variability of pathway genes. We assume that the eigenvalues of previously defined pathways capture biologically relevant quantities, and we develop a test for biologically meaningful changes in the eigenvalues between classes. This test reflects important and often ignored aspects of pathway behavior and provides a useful complement to traditional pathway analyses.