Sim Ah Siew, Salonikas Chris, Naidoo Daya, Wilcken David Emil Leon
Cardiovascular Genetics Laboratory, Department of Medicine, Prince of Wales Hospital, High Street, Randwick, Sydney, NSW Australia 2031.
J Chromatogr B Analyt Technol Biomed Life Sci. 2003 Mar 5;785(2):337-44. doi: 10.1016/s1570-0232(02)00956-x.
Measurement of malondialdehyde (MDA) is an important contribution to the assessment of oxidative stress. We report a sensitive and reproducible high-performance liquid chromatography (HPLC) method for measurement of plasma MDA in the assessment of lipid peroxidation. Using methyl malondialdehyde (Me-MDA) as an internal standard with reversed-phase HPLC and UV detection and derivatisation with 2,4 dinitrophenylhydrazine (DNPH), we obtained maximum MDA values with 60-min incubation of 10% plasma with 1 M NaOH at 60 degrees C. The dilution of the plasma and a longer incubation time in the alkaline hydrolysis step greatly improved recovery of MDA from its bound form. Ratios of peak height of MDA/Me-MDA were linear over a range of 0-100 microM with correlation coefficients >0.99. The recovery was 88.5%. Within and between run variations were <4 and <7%, respectively. The mean MDA value measured in 20 healthy volunteers was 13.8 microM (+/-1.32).
丙二醛（MDA）的测定对氧化应激评估具有重要意义。我们报告了一种灵敏且可重复的高效液相色谱（HPLC）方法，用于在脂质过氧化评估中测定血浆MDA。使用甲基丙二醛（Me-MDA）作为内标，采用反相HPLC和紫外检测，并与2,4-二硝基苯肼（DNPH）进行衍生化反应，我们在60℃下将10%血浆与1 M NaOH孵育60分钟后获得了最大MDA值。血浆稀释以及在碱性水解步骤中延长孵育时间极大地提高了MDA从其结合形式中的回收率。MDA/Me-MDA的峰高比在0 - 100 microM范围内呈线性，相关系数>0.99。回收率为88.5%。批内和批间变异分别<4%和<7%。在20名健康志愿者中测得的平均MDA值为13.8 microM（±1.32）。
