Dills R L, Shen D D
Department of Pharmaceutics, School of Pharmacy, University of Washington, Seattle 98195, USA.
J Chromatogr B Biomed Sci Appl. 1997 Mar 7;690(1-2):139-52. doi: 10.1016/s0378-4347(96)00405-7.
Analysis of the branched, medium-chain fatty acid anticonvulsant, valproic acid, and its unsaturated metabolites by gas chromatography with electron-capture detection suffered from background interference caused by the derivatizing reagent pentafluorobenzyl bromide. Background was reduced by keeping the derivatization anhydrous, using an inert solvent, minimizing the amount of pentafluorobenzyl bromide, using hypernucleophilic bases and displacing the derivatization solvent with isooctane. However, these strategies proved difficult to reproduce. Post-derivatization clean-up with HPLC was much more reliable and provided sufficient sensitivity for the analysis of extracts of plasma and brain homogenate. The assay was validated for plasma and brain samples from humans, rats and mice.
采用带有电子捕获检测的气相色谱法分析支链中链脂肪酸抗惊厥药丙戊酸及其不饱和代谢物时，会受到衍生化试剂五氟苄基溴引起的背景干扰。通过保持衍生化无水、使用惰性溶剂、尽量减少五氟苄基溴的用量、使用超亲核碱并用异辛烷置换衍生化溶剂来减少背景。然而，这些策略难以重复实施。用高效液相色谱法进行衍生化后净化更为可靠，并且为血浆和脑匀浆提取物的分析提供了足够的灵敏度。该测定方法在人、大鼠和小鼠的血浆和脑样本中得到了验证。
