De Zwart L L, Venhorst J, Groot M, Commandeur J N, Hermanns R C, Meerman J H, Van Baar B L, Vermeulen N P
Leiden-Amsterdam Center for Drug Research (LACDR), Department of Pharmacochemistry, Amsterdam, Netherlands.
J Chromatogr B Biomed Sci Appl. 1997 Jul 4;694(2):277-87. doi: 10.1016/s0378-4347(97)00144-8.
One of the major processes that occur as a result of radical-induced oxidative stress is lipid peroxidation (LPO). Degradation of lipid peroxides results in various products, including a variety of carbonyl compounds. In the present study eight different lipid degradation products, i.e., formaldehyde, acetaldehyde, acetone, propanal, butanal, pentanal, hexanal and malondialdehyde were identified and measured simultaneously and quantitatively in rat urine after derivatization with O-(2,3,4,5,6-pentafluorbenzyl)hydroxylamine hydrochloride, extraction with heptane and using gas chromatography-electron-capture detection (GC-ECD). The identity of the respective oximes in urine was confirmed by gas chromatography-negative ion chemical ionization mass spectrometry (GC-NCI-MS). Simultaneously measured standard curves were linear for all oxime-products and the detection limits were between 39.0 +/- 5.3 (n=9) and 500 +/- 23 (n=9) fmol per microl injected sample. Recoveries of all products from urine or water were 73.0 +/- 5.2% and higher. In urine of CCl4-treated rats an increase in all eight lipid degradation products in urine was found 24 h following exposure. ACON showed the most distinct increase, followed by PROPA, BUTA and MDA. It is concluded that the rapid, selective and sensitive analytical method based on GC-ECD presented here is well suited for routine measurement of eight different lipid degradation products. These products appear to be useful as non-invasive biomarkers for in vivo oxidative stress induced in rats by CCl4.
自由基诱导的氧化应激所引发的主要过程之一是脂质过氧化（LPO）。脂质过氧化物的降解会产生多种产物，包括各种羰基化合物。在本研究中，用盐酸O-（2,3,4,5,6-五氟苄基）羟胺进行衍生化、用庚烷萃取并采用气相色谱-电子捕获检测（GC-ECD）法，同时对大鼠尿液中的八种不同脂质降解产物，即甲醛、乙醛、丙酮、丙醛、丁醛、戊醛、己醛和丙二醛进行了定性和定量测定。通过气相色谱-负离子化学电离质谱（GC-NCI-MS）法确认了尿液中相应肟类化合物的身份。所有肟类产物的同时测量标准曲线均呈线性，检测限在每微升进样样品39.0±5.3（n = 9）至500±23（n = 9）飞摩尔之间。所有产物从尿液或水中的回收率均在73.0±5.2%及以上。在四氯化碳处理的大鼠尿液中，暴露24小时后发现所有八种脂质降解产物均增加。丙烯醛显示出最明显的增加，其次是丙醛、丁醛和丙二醛。结论是，本文介绍的基于GC-ECD的快速、选择性和灵敏的分析方法非常适合对八种不同脂质降解产物进行常规测量。这些产物似乎可用作四氯化碳诱导大鼠体内氧化应激的非侵入性生物标志物。
