HPLC method development for quantification of doxorubicin in cell culture and placental perfusion media

Mansi Shah, Luke Bourner, Shariq Ali, Sanaalarab Al-Enazy, Menatallah M. Youssef, Morgan Fisler, Erik Rytting

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Assessment of drug transport across the placenta is important in understanding the effect of drugs on placental and fetal health. These phenomena can be studied in both in vitro cell lines and ex vivo placental perfusions. We have successfully developed a sensitive yet simple high performance liquid chromatography (HPLC) method coupled with fluorescence detection to determine the concentration of doxorubicin (DXR) in cell culture media for transport studies in human trophoblast cells (BeWo, b30 clone) and in fetal media for placental perfusion experiments. The method was developed based on a protein precipitation technique and was validated in both media types for linearity, intra-day, and inter-day precision and accuracy. The relationship of peak area to concentration was linear with R2values of 0.99 or greater obtained over the concentration range of 1.5 to 15,000 ng/mL. Despite the high concentrations of albumin in fetal perfusion media (30 mg/mL), the lower limits of detection and quantification for DXR were found to be 1.5 and 5 ng/mL, respectively. This analytical method may be used to study the transport of DXR across BeWo cells and human placenta during placental perfusion studies.

Original languageEnglish (US)
Article number9
JournalSeparations
Volume5
Issue number1
DOIs
StatePublished - Mar 2018

Keywords

  • Analytical method
  • Anthracycline
  • BeWo cell culture media
  • Doxorubicin
  • Fluorescence
  • High performance liquid chromatography
  • Perfusion
  • Placenta

ASJC Scopus subject areas

  • Analytical Chemistry
  • Filtration and Separation

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