Determination of Methotrexate in Human Serum by HPLC / 中国药房

OBJECTIVE:To determinate the serum concentration of methotrexate by HPLC. METHODS:The samples were separated on a Waters C18 column at a column temperature of 30 ℃. The mobile phase consisted of acetonitrile-phosphate buffer(pH 7.2,12∶88) at a flow rate of 1.0 mL?min-1. The detection wavelength was set at 303 nm. RESULTS:The linear range of methotrexate was 0.5～80 ?g?mL-1(r=0.999 4). The methodological recovery rates were all above 90%; the intraday RSD was 3.03 %～3.52 % and the inter-day RSD was 2.57%～4.05%. CONCLUSION:The method is proved to be simple,rapid,accurate and sensitive,and it is suitable for the determination of serum concentration and pharmacokinetic study of methotrexate.