마우스 혈장과 조직에서의 doxorubicin 측정 HPLC-MS/MS 방법

Abstract

Doxorubicin (DXR) is a type of anti-cancer drug called an “anthracycline glycoside”. It works by impairing DNA synthesis, a crucial feature of cell division, and thus is able to target rapidly dividing cells. Doxorubicin is a very serious anti-cancer medication with definite potential to do great harm as well as great good. A liquid chromatography-tandem mass spectroscopy (LC–MS/MS) method was developed to identify and quantify DXR in small-volume biological samples. After the addition of internal standard (IS, 5 μL of 1 μM/ml daunorubicin methanol solution) into the serum sample, the drug and IS were extracted by methanol. Following vortex for a 1 min and centrifugation at 15,000g for 10 min the organic phase was transferred and evaporated under a vacuum. The residue was reconstituted with 350 μL of mobile phase and 10 μL was injected into C18 column with mobile phase composed of 0.05 M ammonium acetate (0.1 M acetic acid adjusted to pH 3.5) and acetonitrile (40:60, v/v). The flow rate was kept constant at 350 μL/min. The ions were quantified in the multiple reaction mode (MRM), using positive ions, on a triple quadrupole mass spectrometer. The lower limits of quantification for Doxorubicin in plasma and small tissues were approximately 0.5 ng/mL and 0.5 ng/mL respectively. Intra- and inter-assay accuracy (% of nominal concentration) and precision (% CV) for all analytes were within 15%, respectively.