Distribution of platinum between nuclear and cytosolic fractions: can subcellular fractionation be performed quantitatively?

Celina Støving Dam, Ian Henry Lambert, Bente Gammelgaard, Stefan Stürup*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

6 Citations (Scopus)

Abstract

The feasibility of quantitatively tracking platinum, derived from platinum-based compounds, during subcellular fractionation was studied. Cisplatin-exposed murine Ehrlich Lettré Ascites cells were fractionated into cytosolic and crude nuclear fractions. The latter was subsequently purified. Every residue and fraction produced during the fractionation procedure were collected and the platinum content determined by inductively coupled plasma mass spectrometry. Western blotting verified that the nuclear and cytosolic fractions were pure. It was found that 18% of platinum taken up by the cells was located in the nuclear fraction while 66% was located in the cytosolic fraction. Accumulated uncertainty originating from invariable sample characteristics and giving fraction purity priority had a negative effect on platinum recovery. Thus, overall 81% (n = 3, RSD = 3.4%) of the platinum taken up by the cells was recovered in the residues and final fractions. In conclusion, a reliable intracellular localization and quantitation of platinum following administration of Cisplatin can be determined by application of the method.

Original languageEnglish
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume165
Pages (from-to)82-89
Number of pages8
ISSN0731-7085
DOIs
Publication statusPublished - 2019

Keywords

  • Cisplatin
  • Inductively coupled plasma mass spectrometry
  • Quantitative
  • Subcellular distribution
  • Subcellular fractionation

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