Steijger O M, Lingeman H, Brinkman U A, Holthuis J J, Smilde A K, Doornbos D A
Department of Analytical Chemistry, Free University, Amsterdam, Netherlands.
J Chromatogr. 1993 May 19;615(1):97-110. doi: 10.1016/0378-4347(93)80295-f.
N-(4-Aminobutyl)-N-ethylisoluminol was used for labelling of carboxylic acids. The derivatization reaction was carried out with 1-hydroxybenzotriazole as pre-activator of the carboxylic acid function and N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide as the coupling reagent. Optimum conditions for the derivatization were determined by using factorial design analysis, with ibuprofen as the test compound. Chemiluminescence detection was carried out using a post-column on-line electrochemical hydrogen peroxide generation system and the addition of microperoxidase as the catalyst. The detection limit of derivatized ibuprofen in human saliva was 0.7 ng per 0.5 ml of saliva, with a recovery of 96.1 +/- 1.3%. The method was linear over at least three decades (2.5 ng to 2.5 micrograms) and the repeatability was satisfactory (R.S.D. = 5.2% at the 25 ng level; n = 4).
N-(4-氨基丁基)-N-乙基异鲁米诺用于羧酸的标记。衍生化反应以1-羟基苯并三唑作为羧酸官能团的预活化剂，N-乙基-N'-(3-二甲基氨基丙基)碳二亚胺作为偶联试剂进行。通过因子设计分析，以布洛芬作为测试化合物，确定了衍生化的最佳条件。化学发光检测使用柱后在线电化学过氧化氢生成系统，并添加微过氧化物酶作为催化剂。衍生化布洛芬在人唾液中的检测限为每0.5毫升唾液0.7纳克，回收率为96.1±1.3%。该方法至少在三个数量级（2.5纳克至2.5微克）范围内呈线性，重复性良好（25纳克水平时相对标准偏差为5.2%；n = 4）。
