[采用高压液相色谱法（HPLC）对接受甲氨蝶呤（MTX）治疗的患者进行血清甲氨蝶呤（MTX）和7-羟基甲氨蝶呤浓度监测，并比较HPLC法与荧光偏振免疫分析法（FPIA）测定的血清MTX水平]

[Serum monitoring of methotrexate (MTX) and 7-hydroxymethotrexate concentrations in patients treated with MTX using high-pressure liquid chromatography (HPLC) and comparison of serum MTX levels between HPLC method and fluorescence polarization immunoassay (FPIA)].

作者信息

Sakamoto S, Matsuhisa T, Miyoshi K, Naruse R, Urayama H, Shimokawa T, Okabe K, Toki H, Motoi M, Moriwaki S

机构信息

Dept. of Pharmacy, Shikoku Cancer Center Hospital.

出版信息

Gan To Kagaku Ryoho. 1991 Jun;18(7):1119-25.

PMID:2053771

Abstract

In order to measure simultaneously the serum levels of methotrexate (MTX) and its major metabolite, 7-hydroxymethotrexate (7-OH-MTX), in samples obtained from patients treated with MTX, we have investigated the reversed-phase high-pressure liquid chromatographic assay using ion-pairing reagents. The mobile phase consisted of 77.5% solution of 0.005M tetrabutylammonium and 22.5% acetonitrile. SEP-PAK C18 Cartridges were used for the precolumn. The detectable range of MTX and 7-OH-MTX were 0.02-0.03 and 1.0 mumol/l respectively. A significant positive correlation was observed (r = 0.983) between FPIA and HPLC methods. Serum MTX levels with FPIA were significantly (p less than 0.05) higher than those of HPLC method. The serum 7-OH-MTX levels at 24 hr and 48 hr were 4.857 +/- 1.383 (n = 10) and 1.835 +/- 0.286 (n = 6) mumol/l respectively with the dosage of 400 mg/m2. The serum 7-OH-MTX levels at 48 hr were 6.254 +/- 3.053 mumol/l (n = 5) with the dosage of 3,000 mg. The serum half lives of MTX and 7-OH-MTX were 8.05 +/- 1.03 (n = 4) and 14.8 +/- 1.35 (n = 6) hours respectively between 24 hr and 48 hr after administration. The T1/2 7-OH-MTX/MTX ratio was 1.8. Percent cross-reactivity of 7-OH-MTX with concentrations ranging from 1-10 mumol/l were 0.6-2.0% by FPIA. However, patients' serum levels of 7-OH-MTX were 15-85 times (n = 21) higher than those of MTX. MTX levels of containing both MTX and 7-OH-MTX (7-OH-MTX/MTX ratio was 50/1) were significantly higher than those of containing MTX alone by FPIA.

摘要

为了同时测定接受甲氨蝶呤（MTX）治疗的患者样本中甲氨蝶呤（MTX）及其主要代谢产物7 - 羟基甲氨蝶呤（7 - OH - MTX）的血清水平，我们研究了使用离子对试剂的反相高效液相色谱法。流动相由0.005M四丁基铵的77.5%溶液和22.5%乙腈组成。SEP - PAK C18柱用于柱前处理。MTX和7 - OH - MTX的可检测范围分别为0.02 - 0.03和1.0μmol/L。荧光偏振免疫分析（FPIA）法与高效液相色谱（HPLC）法之间观察到显著正相关（r = 0.983）。FPIA法测定的血清MTX水平显著高于HPLC法（p < 0.05）。剂量为400mg/m²时，24小时和48小时的血清7 - OH - MTX水平分别为4.857±1.383（n = 10）和1.835±0.286（n = 6）μmol/L。剂量为3000mg时，48小时的血清7 - OH - MTX水平为6.254±3.053μmol/L（n = 5）。给药后24小时至48小时之间，MTX和7 - OH - MTX的血清半衰期分别为8.05±1.03（n = 4）和14.8±1.35（n = 6）小时。7 - OH - MTX/MTX的T1/2比值为1.8。FPIA法测定，浓度范围为1 - 10μmol/L的7 - OH - MTX的交叉反应率为0.6 - 2.0%。然而，患者血清中7 - OH - MTX水平比MTX高15 - 85倍（n = 21）。FPIA法测定，同时含有MTX和7 - OH - MTX（7 - OH - MTX/MTX比值为50/1）的MTX水平显著高于仅含MTX的样本。