Validation of a HPLC MS/MS Method for Determination of Doxorubicin in Mouse Serum and its Small Tissues

마우스 혈장과 조직에서의 doxorubicin 측정 HPLC-MS/MS 방법

  • Park, Jung-Sun (Department of Pharmacology and Cardiovascular Research Center, Medical School, Chonbuk National University) ;
  • Kim, Hye-Kyung (Clinical Trial Center for Functional Foods, Chonbuk Hospital) ;
  • Lee, Hye-Won (Clinical Trial Center for Functional Foods, Chonbuk Hospital) ;
  • Lee, Mi-Hyun (Clinical Trial Center for Functional Foods, Chonbuk Hospital) ;
  • Kim, Hyun-Gi (Department of Pharmacology and Cardiovascular Research Center, Medical School, Chonbuk National University) ;
  • Chae, Soo-Wan (Department of Pharmacology and Cardiovascular Research Center, Medical School, Chonbuk National University) ;
  • Chae, Han-Jung (Department of Pharmacology and Cardiovascular Research Center, Medical School, Chonbuk National University)
  • 박정선 (전북대학교 의과대학 약리학교실 및 심혈관연구소) ;
  • 김혜경 (전북대학교병원 기능성식품 임상시험지원센타) ;
  • 이혜원 (전북대학교병원 기능성식품 임상시험지원센타) ;
  • 이미현 (전북대학교병원 기능성식품 임상시험지원센타) ;
  • 김현기 (전북대학교 의과대학 약리학교실 및 심혈관연구소) ;
  • 채수완 (전북대학교 의과대학 약리학교실 및 심혈관연구소) ;
  • 채한정 (전북대학교 의과대학 약리학교실 및 심혈관연구소)
  • Published : 2006.06.30

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, $5{\mu}L\;of\;1{\mu}M/ml$ daunorubicin methanol solution) into the serum sample, the drug and IS were extracted by methanol. Following vortex for a 1min and centrifugation at 15,000g for 10 min the organic phase was transferred and evaporated under a vacuum. The residue was reconstituted with $350{\mu}L$ of mobile phase and $10{\mu}L$ was injected into C18 column with mobile phase composed of 0.05M 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 $350{\mu}L/min$. 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.

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