Determination of serum concentration of methotrexate in children with acute lymphoblastic leukemia by HPLC and application practice research / 中国生化药物杂志

ABSTRACT

Objective To determine serum concentration of methotrexate in children with acute lymphoblastic leukemia by HPLC and explore the application practice.Methods The separation was performed in a Wondasil C18-WR column (4.6 mm ×150 mm, 5 μm) with a mobile phase of methanol-0.15%phosphoric acid solution(2179 ) and determined at 306 nm.The sample of serum was centrifuged after protein precipitation with perchloric acid.Detected results for children to establish monitoring files.ResuIts The linearity was well at 0.02 ~50 μg/mL of methotrexate ( r =0.9990,n=6).The average recovery was 96.50%, RSD of within day and between days were less than 15%.Established monitoring files for six children and after 3 courses of treatment, the blood concentration of methotrexate in 4 children were small and they were in a safe range , which could reduce the frequency of monitoring.ConcIusion This method is simple, sensitive and accurate for methotrexate detection in serum.The children with stable concentration and within safety range, can reduce monitioring frequency.It could bring the children less pain and relieve the burden of family.