7-羟基甲氨蝶呤代谢物对甲氨蝶呤测定临床相关性的影响。
The influence of 7-OH methotrexate metabolite on clinical relevance of methotrexate determination.
作者信息
Klapkova Eva, Kukacka Jiri, Kotaska Karel, Suchanska Iveta, Urinovska Romana, Prusa Richard
机构信息
Department of Clinical Biochemistry and Pathobiochemistry, Charles University, 2nd Faculty of Medicine and Faculty Hospital Motol, Prague, Czech Republic.
出版信息
Clin Lab. 2011;57(7-8):599-606.
BACKGROUND
A modified high performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of methotrexate (MTX) and its main metabolite 7-hydroxymethotrexate (7-OHMTX) and compared to the immunochemical fluorescence polarization immunoassay (FPIA2) determination of methotrexate.
METHODS
Methotrexate was determined by HPLC with UV detection at 303 nm after precipitation of proteins with trichloroacetic acid. Fluorescence polarization immunoassays (FPIA2) of methotrexate were performed on the TDx FLx Immunoassay Analyzer.
RESULTS
Our data indicate good correlation between methotrexate levels > 1 micromol/L determined by HPLC and FPIA2. (r = 0.94, Spearman correlation coefficient). However, concentrations of methotrexate < 1 micromol/L measured by fluorescence polarization immunoassay were overestimated.
CONCLUSIONS
The concentration of MTX < 1 micromol/L are overestimated due to the cross reactivity with metabolites 7-OHMTX and 2,4-diamino-N10-methylpteroic acid (DAMPA). The cross reaction may affect the therapy and lead to relapse in children with acute lymphoblastic leukemia.
背景
已开发出一种改良的高效液相色谱（HPLC）方法，用于同时测定甲氨蝶呤（MTX）及其主要代谢产物7-羟基甲氨蝶呤（7-OHMTX），并与甲氨蝶呤的免疫化学荧光偏振免疫测定法（FPIA2）进行比较。
方法
用三氯乙酸沉淀蛋白质后，通过在303nm处进行紫外检测的HPLC法测定甲氨蝶呤。在TDx FLx免疫分析仪上进行甲氨蝶呤的荧光偏振免疫测定（FPIA2）。
结果
我们的数据表明，通过HPLC和FPIA2测定的甲氨蝶呤水平>1微摩尔/升之间具有良好的相关性。（r = 0.94，斯皮尔曼相关系数）。然而，通过荧光偏振免疫测定法测得的甲氨蝶呤浓度<1微摩尔/升被高估了。
结论
由于与代谢产物7-OHMTX和2,4-二氨基-N10-甲基蝶酸（DAMPA）的交叉反应，MTX浓度<1微摩尔/升被高估。这种交叉反应可能会影响治疗，并导致急性淋巴细胞白血病患儿复发。
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