van Tellingen O, Beijnen J H, van der Woude H R, Bruning P F, Nooyen W J
Department of Clinical Chemistry and Clinical Oncology, Netherlands Cancer Institute, Amsterdam.
J Chromatogr. 1990 Jul 13;529(1):135-43. doi: 10.1016/s0378-4347(00)83814-1.
10-Ethyl-10-deaza-aminopterin (10-EdAM) is a novel folic acid antimetabolite currently being tested in phase II clinical trials. We have developed an isocratic high-performance liquid chromatographic method for the quantification of 10-EdAM and metabolites in plasma. Solid-phase extraction was used for sample clean-up. Adequate accuracy was obtained without the use of an internal standard. Fluorometric detection with excitation at 243 nm and emission at 488 nm was used for accurate quantification of samples containing small amounts of drug or metabolites (2.0-4.0 nM, depending on the compound). Ultraviolet detection at 350 nm was only applicable for the analysis of plasma concentrations of 10-EdAM exceeding 50 nM. The usefulness of the assay was demonstrated by the results obtained in a pharmacokinetic study. The assay could separate the parent compound from seven identified and two unknown products.
10-乙基-10-脱氮氨基蝶呤（10-EdAM）是一种新型叶酸抗代谢物，目前正在进行II期临床试验。我们开发了一种等度高效液相色谱法，用于定量血浆中的10-EdAM及其代谢物。采用固相萃取法进行样品净化。在不使用内标物的情况下获得了足够的准确度。使用激发波长为243nm、发射波长为488nm的荧光检测法对含有少量药物或代谢物（2.0-4.0 nM，取决于化合物）的样品进行准确定量。350nm处的紫外检测仅适用于分析血浆中浓度超过50 nM的10-EdAM。药代动力学研究结果证明了该测定方法的实用性。该测定方法能够将母体化合物与七种已鉴定的产物和两种未知产物分离。
