Holley A E, Walker M K, Cheeseman K H, Slater T F
Brunel University, Department of Biology and Biochemistry, Uxbridge, Middlesex, UK.
Free Radic Biol Med. 1993 Sep;15(3):281-9. doi: 10.1016/0891-5849(93)90075-6.
A modified method was developed to measure nM levels of a range of n-alkanals and hydroxyalkenals in biological samples such as blood plasma and tissue homogenates and also in Folch lipid extracts of these samples. Butylated hydroxytoluene (BHT) and desferrioxamine (Desferal) were added to samples to prevent artifactual peroxidation. Aldehydes were reacted with 1,3-cyclohexanedione (CHD), cleaned up by solid-phase extraction on a Sep-Pak C18 cartridge and the fluorescent decahydroacridine derivatives resolved by reverse-phase high-performance liquid chromatography (HPLC) with gradient elution. A wider range of aldehydes was detected in lipid extracts of plasma and liver homogenate compared to whole (unextracted) samples. Human plasma contained nM levels of acetaldehyde, propanal, butanal, pentanal, hexanal, and heptanal. 4-Hydroxynonenal (0.93 nmol/g) and alkanals with two to six carbons (up to 7.36 nmol/g) were detected in rat liver. Recovery of aldehydes added to whole plasma or to lipid extracts of plasma was dependent on carbon chain length, varying from 95% for acetaldehyde to 8% for decanal. Recovery from biological samples was significantly less than that of standards taken through the Sep-Pak clean-up procedure, suggesting that aldehydes can bind to plasma protein and lipid components.
已开发出一种改良方法，用于测量生物样品（如血浆和组织匀浆）以及这些样品的福氏脂质提取物中一系列正构烷醛和羟基烯醛的纳摩尔水平。向样品中添加丁基羟基甲苯（BHT）和去铁胺（去铁敏）以防止人为的过氧化反应。醛与1,3 - 环己二酮（CHD）反应，通过Sep - Pak C18柱进行固相萃取净化，然后通过反相高效液相色谱（HPLC）梯度洗脱分离荧光十氢吖啶衍生物。与完整（未提取）样品相比，在血浆和肝脏匀浆的脂质提取物中检测到更广泛的醛类。人血浆中含有纳摩尔水平的乙醛、丙醛、丁醛、戊醛、己醛和庚醛。在大鼠肝脏中检测到4 - 羟基壬烯醛（0.93 nmol/g）和含有2至6个碳的正构烷醛（高达7.36 nmol/g）。添加到全血浆或血浆脂质提取物中的醛的回收率取决于碳链长度，从乙醛的95%到癸醛的8%不等。从生物样品中的回收率明显低于通过Sep - Pak净化程序的标准品回收率，这表明醛可以与血浆蛋白和脂质成分结合。
