UPLC法测定抗肿瘤药物甲氨蝶呤血药浓度

目的临床上建立一种快速、准确测定肿瘤患儿血浆中甲氨蝶呤浓度方法。方法取肿瘤患儿血浆250μL,加入7%高氯酸200μL,涡旋1 min,室温高速离心5 min(15 000×g),取上清液5μL进样分析。色谱条件:色谱柱为BEH C18柱(50mm×2.1 mm,1.7μm);流速为0.3 mL·min-1;流动相为乙腈-40 mol·L-1磷酸二氢钾缓冲液(91∶9,pH=4.5);柱温为30℃;进样量:5μL;检测波长为304 nm。结果血浆中MTX色谱峰分离良好,无干扰,曲线为y=41 729x-810.29,R2=0.9 997,回收率在90%¹10%之间。结论 UPLC操作简便,适用于血浆中MTX快速监测。

Determination of Plasma Concentration of Methotrexate by UPLC

Objective To establish a rapid and accurate method to determine the plasma concentration of methotrexate in children with tumor. Methods 0.25μL plasma specimen taken from tumorous children was extracted with 7%perchloric acid and swirled 1 min, then it was high-speed centrifuged 5 min （15 000μg）. 5μL supernate was injected in UPLC assay for analysis. The analytical column was BEH C18 （50 mmμ2.1 mm, 1.7μm）, the flow rate was 0.3 mL·min-1, and the mobile phase was the buffer of acetonitrile-40 mol·L-1 and NaH2PO4 （91： 9, pH = 4.5）. The column temperature was 30 ℃, the sample size was 5 μL, and the wavelength was 304 nm. Results The chromatographic peak of methotrexate in plasma separated good without interference. The curve was y=41 729x-810.29, R2=0.9 997, the recycle rate was between 90%and 100%. Conclusions UPLC is easy to operate and fit for fast determination of MTX in human plasma.