Determination of Cisplatin using Hydrophilic Interaction Liquid Chromatography (HILIC) Coupled with Inductively Coupled Plasma Mass Spectrometry (ICP-MS): A Study on Cisplatin’s Retention Mechanisms in HILIC and Uptake and Intracellular Decay Kinetics in in-vitro Grown Cells
Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Cisplatin (cis-diamminedichloroplatinum(II), CDDP) is widely used for the treatment of several solid tumors. The antitumor property of cisplatin is generally proved that the covalent binding of cisplatin to DNA distorts the DNA structure, which leads to the death of the cancer cells. The obstacle for chemotherapy is that tumors can acquire resistance to cisplatin and induce the severe side effects associated with cisplatin treatment. How cisplatin enters the cells and metabolizes in cytoplasm is not completely elucidated. The purpose of this project was to study the kinetics of cellular uptake of cisplatin and decay of intracellular cisplatin in in-vitro grown cells. Another aim was to investigate the retention mechanism of cisplatin on a zwitter-ionic hydrophilic interaction liquid chromatography (ZIC®-HILIC) column.
HILIC coupled on-line to inductively coupled plasma mass spectrometry was applied to separate and quantify cisplatin content in cell lysates from malignant melanoma cells exposed to cisplatin. The cells were lysed by using a ceramic bead method.
Both sensitive and resistant cells gained the maximum total platinum uptake after 60 minutes. The sensitive cells accumulate more total-Pt but not more free cisplatin than resistant cells.The half-life of cisplatin in resistant cells exposed to 20 and 100μM cisplatin was found to be 17 and 14 minutes, respectively. The results also indicated that the interaction of cisplatin with ZIC-HILIC stationary phase was likely to be hydrogen bonding interaction.
Place, publisher, year, edition, pages
2012. , 46 p.
IdentifiersURN: urn:nbn:se:umu:diva-56554OAI: oai:DiVA.org:umu-56554DiVA: diva2:535739
UppsokPhysics, Chemistry, Mathematics