hplc analysis of DoxorubicinDid you know that the cancer drug Doxorubicin is derived from a bacterial species by chemical semisynthesis and is used to treat a wide range of cancers? (Link & source Wikipedia). A pegylated liposomal formulation of doxorubicin is used in cancer treatment and it is essential to establish a fast and robust LC-MS/MS method for separation and determination of liposomal and non-liposomal (free) doxorubicin present in circulating blood.

The method is described in Application Note 20937, Separation and Determination of Liposomal and Non-Liposomal (Free) Doxorubicin from Human Plasma by SPE and LC-MS/MS, (downloadable PDF), and features the use of one of our fritless, solid-phase extraction (SPE) cartridges (Thermo Scientific SOLA HRP SPE cartridge) specifically designed for bioanalysis. These SPE cartridges provide higher levels of reproducibility, higher levels of extract cleanliness, and reduced solvent requirements.

Following the selective retention of liposomal (encapsulated) doxorubicin and non-liposomal (free) doxorubicin on the SPE cartridge, researchers at our Asia Pacific Center of Excellence, located in the city of Ahmedabad, India, treated the liposomal (encapsulated) fraction with a protein precipitation methodology to disrupt the liposome with subsequent SPE clean up. This was followed by separation on a fast, high-resolution HPLC column (Thermo Scientific Accucore C18 HPLC column) and one of our rapid-separation HPLC systems (Thermo Scientific Dionex UltiMate 3000RS system). For the detection, one of ourtriple quadrupole mass spectrometers (Thermo Scientific TSQ Vantage triple quadrupole mass spectrometer) was used.

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In their results, the researchers achieved a limit of quantitation of 1 ng/mL of doxorubicin in plasma; the extraction recovery and matrix interference were found within the limits of acceptance generally applied to bioanalytical methods; good accuracy and precision was observed for both liposomal (encapsulated) and non-liposomal (free) doxorubicin at each QC level.

The stunning image is of the structures of doxorubicin and the internal standard, daunorubicin.

Additional Resources

  • Our recently released Chromatography Solution Online Center features many useful and complimentary chromatography tools which can help speed up your analysis. The site is updated on a monthly basis; therefore, do check out the Archives section to see what was previously featured.

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