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KMID : 0941820060160010023
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Korean Journal of Clinical Pharmacy
2006 Volume.16 No. 1 p.23 ~ p.27
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Validation of HPLC MS/MS Method for Determination of Doxorubicin in Mouse Serum and its Small Tissues
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Park Jung-Sun
Kim Hye-Kyung
Lee Hye-Won
Lee Mi-Hyun
Kim Hyun-Gi
Chae Soo-Wan
Chae Han-Jung
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Abstract
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Doxorubicin (DXR) is a type of anti-cancer drug called an "anthracycline glycoside". It words 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 liq- uid 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, 5μL of 1μM/ml daunorubicin methanol solution) into the serum sample, the drug and IS were extracted by methanol. Following vortex for 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 350μL of mobile phase and 10μL was injected into C18 column with moile 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 350μL /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 ana- lytes were within 15%, respectively.
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KEYWORD
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doxorubicin, daunorubicin, HPLC-MS/MS, validation
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