期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 398, 期 5, 页码 2183-2190出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-010-4152-3
关键词
Capillary electrophoresis; Cerebrospinal fluid; Metabolites; Methotrexate; Triple-stacking
资金
- Center of Excellence for Environmental Medicine [KMU-EM-98-1-2]
- National Science Council of Taiwan
- Kaohsiung Medical University
We establish a triple-stacking capillary electrophoresis (CE) separation method to monitor methotrexate (MTX) and its eight metabolites in cerebrospinal fluid (CSF). Three stacking methods with different mechanisms were combined and incorporated into CE separation. Complete stacking and sharp peaks were achieved. Firstly, the optimized buffer (60 mM phosphate containing 15% THF and 100 mM SDS) was filled into the capillary, which was followed by the higher conductivity buffer (100 mM phosphate, 2 psi for 45 s). The analytes extracted from CSF were injected at 2 psi for 99.9 s, which provided long sample zones and pH junction for focusing. Finally, the stacking step was performed by sweeping, and separation was achieved by micellar electrokinetic chromatography. The results of the linear regression equations indicated high linearity (r a parts per thousand yenaEuro parts per thousand 0.9981) over the range of 0.5-7 mu M. In intra- and inter-batch results, all data of RSD and RE were below 11%, indicating good precision and accuracy of this method. The LODs (S/N = 3) were 0.1 mu M for MTX, 7-hydroxymethotrexate (7-OHMTX) and MTX-polyglutamates (MTX-(Glu) (n, n = 2-5)), 0.2 mu M for MTX-(Glu)(6), and 0.3 mu M for 2,4-diamino-N (10)-methylpteroic acid (DAMPA) and MTX-(Glu)(7). Our method was implemented for analysis of MTX and its metabolites in the CSF, and could be used for evaluation of its curative effects of acute lymphoblastic leukemia patients. The data were also confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The results showed good coincidence.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据