title: A reversed-phase high performance liquid chromatography method for quantification of methotrexate in cancer patients serum.
abstract: A simple, rapid and sensitive reversed-phase high performance liquid chromatography (HPLC) method has been developed for the determination of methotrexate in human serum. After deproteinization of the serum with 40% silver nitrate solution, methotrexate and internal standard (IS) were separated on a reversed-phase column with a mobile phase consisting of 10mM sodium phosphate buffer (pH6.40)-methanol (78:22%, v/v) and ultraviolet detection at 310nm. The linearity is evaluated by a calibration curve in the concentration range of 0.05-10.0μg/mL and presented a correlation coefficient of 0.9995. The absolute recoveries were 97.52±3.9% and 96.87±3.7% for methotrexate and ferulic acid (internal standard), respectively. The intra- and inter-day precision were less 6.19 and 5.89%, respectively (n=6). The limit of quantitation was 0.02μg/mL and the limit of detection was 0.006μg/mL. The complete analysis was achieved less than 10min with no interference from endogenous components or 22 examined drugs. This method was validated by using serum samples from high-dose methotrexate treated patients with osteosarcoma, breast cancer, acute leukemia and lymphoma. The method was demonstrated to be a simple, rapid and reliable approach in quantification of methotrexate in serum samples from patients with high-dose methotrexate therapy.
authors: Research Department, Affiliated Tumor Hospital of Guangxi Medical University, Nanning, 530021 Guangxi, People's Republic of China. Electronic address: lyd641209@163.com.
authors: Yuan-dong
authors: YD
authors: Li
authors: Research Department, Affiliated Tumor Hospital of Guangxi Medical University, Nanning, 530021 Guangxi, People's Republic of China.
authors: Yan
authors: Y
authors: Li
authors: Research Department, Affiliated Tumor Hospital of Guangxi Medical University, Nanning, 530021 Guangxi, People's Republic of China.
authors: Ning-sheng
authors: NS
authors: Liang
authors: Research Department, Affiliated Tumor Hospital of Guangxi Medical University, Nanning, 530021 Guangxi, People's Republic of China.
authors: Fan
authors: F
authors: Yang
authors: Research Department, Affiliated Tumor Hospital of Guangxi Medical University, Nanning, 530021 Guangxi, People's Republic of China.
authors: Zhi-peng
authors: ZP
authors: Kuang
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000097:blood*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D008727:Methotrexate / Q000097:blood*; D009369:Neoplasms / Q000097:blood*; D012015:Reference Standards; D015203:Reproducibility of Results
chemical_list: D000964:Antimetabolites, Antineoplastic; D008727:Methotrexate
keywords: HPLC; Method validation; Methotrexate; Serum; Silver nitrate