高效液相色谱法测定人体血浆中甲氨蝶呤浓度方法的改进

Improved high-performance liquid chromatographic determination of methotrexate in human plasma

目的:建立高效液相色谱法测定人体血浆中甲氨蝶呤浓度的方法。方法:血浆样品经处理后,色谱分离采用Zorbax-ODSC18(250mm×4.6mm,5μm)色谱柱,检测波长为302nm,流动相为0.01mol.L-1磷酸盐(用磷酸溶液调pH6.7)-甲醇(80∶20),流速为1.0mL.min-1,柱温为35℃。结果:甲氨蝶呤血药质量浓度在0.2～50mg.L-1(r=0.9997)范围内线性关系良好,最低检测质量浓度为5μg.L-1,提取回收率为96.5%～98.7%,方法回收率为98.2%～101.2%,日内、日间RSD分别为3.62%和4.57%。结论:该方法简单、快速、准确适用于临床甲氨蝶呤浓度监测及药动学研究。

PBKECTIVE To develop a new HPLC method for determination of methotrexate in human plasma. METHODS The Zorbax-ODS C18column （250 mm× 4. 6 mm,5μm） was used. The mobile phase consisted of 0. 01 mol·L^-1 phosphate buffer （adjusted pH to 6. 7 with phosphoric acid ）- methanol （80：20）. The flow rate was 1.0 mL·min^-1 and the column temperature was 35 ℃. The detective wavelength was 302 nrn. RESULTS The linear relation was in the range of 0. 2 - 50 mg · L^-1 （r= 0. 999 7）, The detection limit of the methotrexate was 5μg·L^- 1. The extraction and method recovery were 96. 5G - 98. 7G and 98. 2G - 101.2G respectively. The RSDs of intra-day and inter-day were 3.62G and 4. 57G respectively. CON- CLUSION This method is simple and accurate, and is suitable for determination of methotrexate concentration in human plasma and pharmacokinetis studies.