Autoxidation of brain homogenates from various animals as measured by thiobarbituric acid assay

INTRODUCTION: The TBARS assay has been well recognized for determination of lipid peroxidation and oxidative injury in biological samples including brain homogenates. In general, the homogenates are freshly prepared using rat brains as the tissue sources. In this study, we compared the rates of spontaneous lipid peroxidation in brain homogenates obtained from bovine, canine, hen, rat, and swine. In addition, the influences of lyophilization process and storage time up to six months at -20 degrees C without the freeze-thaw cycle were also determined in the swine brain preparations. METHODS: The standard assay for thiobarbituric acid-reactive substances (TBARS) was performed at 37 degrees C, using spectrophotometry to quantify the malondialdehyde (MDA) content. RESULTS: Rat brain homogenate exhibited the highest autoxidation rate (0.128+/-0.002 microM/min) whereas the bovine brain exhibited the lowest rate (0.032+/-0.001 microM/min). Swine brain homogenate could be kept at -20 degrees C up to 3 months without a significant increase in rate of autoxidation. Lyophilization caused a significant increase in the autoxidation rate of brain homogenate. However, the autoxidation rates of the lyophilized preparation were quite comparable throughout the six-month freezing time. DISCUSSION: Swine brain was a good candidate for tissue source in the TBARS reaction. The homogenate could be kept in the lyophilized form under the storage condition at -20 degrees C without the freeze-thaw cycle in the dark for at least six months.