An excellent pre-column fluorescent derivatization reagent N-acetylhydrazine acridone for the quantitative determination of malondialdehyde was synthesized. Malondialdehyde was derivatized at 80 °C for 30 min in the presence of trichloroacetic acid. The separation of the derivative was performed on an Agilent ZORBAX SB-C18 column in conjunction with gradient elution. The excitation and emission wavelengths were 370 nm and 420 nm, respectively. The developed method demonstrated good linear relationship in the range of 0.02 pmol to 2.5 pmol (r = 0.9998). The calculated limit of detection and limit of quantification were 2.5 fmol and 8.3 fmol, respectively. The analytical precisions of the method were in the range of 1.36-2.27% (intra-day) and 2.36-3.92% (inter-day) respectively. The method was sensitive, specific and simple. It was successfully implemented to analysis the malondialdehyde in rat prostate.Reactive oxygen species (ROS) can cause oxidative stress that is related to cell aging and plays an essential role in the pathogenesis of a variety of illnesses. During oxidative stress, the double bonds of polyunsaturated fatty acids are peroxided by ROS to yield lipid hydroperoxides, which are readily decomposed into some secondary products by several sequential reactions. One of the major secondary products is malondialdehyde (MDA) 1,2 . MDA is reactive toward proteins and nucleic acids. It has been inferred to have mutagenic and cytotoxic effects by generating DNA-protein cross-links. MDA was found elevated in various diseases including cancers 3 , diabetes 4 , cardiovascular diseases 5 and liver diseases 6 , making it an indirect indicator of these diseases. Therefore, it is of great significance to determine its content accurately. However, the accurate determination is extremely difficult because of the lower concentration and many interfering factors existing in biological samples 1 . Therefore, a sensitive and selective method for the determination of MDA is needed.Many methods have been developed for the determination of MDA, mainly including spectrometry 7,8 , gas chromatography-mass spectrometry (GC-MS) 9 , high performance liquid chromatography (HPLC) 10 and liquid chromatography with mass spectrometry (LC-MS) 11,12 . MDA shows weak absorption in the UV-visible region, so it is very difficult for direct determination of biological sample by spectrometric techniques. To overcome this difficulty, derivatization has been adopted by most methods developed in recent years. The most commonly used derivatization reagent is 2-thiobarbituric acid (TBA) that can react with MDA at 100 °C with acid as catalyst, and the reaction product has strong ultraviolet-visible absorption at 533 nm 13 . However, TBA can also react with many other carbonyl compounds existing in biological samples, which would result in considerable overestimation for MDA 14 . Furthermore, the sensitivity of this method is not high enough for accurate determination of trace MDA in biological samples 15 . HPLC with strong separation capability co...