Intracellular drug uptake: a comparison of single cell measurements using ToF-SIMS imaging and quantification from cell populations with LC/MS/MS

Newman, Carla F.; Havelund, Rasmus; Passarelli, Melissa K.; Marshall, Peter S.; Francis, Ian; West, Andy; Alexander, Morgan R.; Gilmore, Ian S.; Dollery, Colin T.

Abstract

ToF-SIMS is a label-free imaging method that has been shown to enable imaging of amiodarone in single rat macrophage (NR8383) cells. In this study, we show that the method extends to three other cell lines relevant to drug discovery: human embryonic kidney (HEK293), cervical cancer (HeLa), and liver cancer (HepG2). There is significant interest in the variation of drug uptake at the single cell level, and we use ToF-SIMS to show that there is great diversity between individual cells and when comparing each of the cell types. These single cell measurements are compared to quantitative measurements of cell-associated amiodarone for the population using LC/MS/MS and cell counting with flow cytometry. NR8383 and HepG2 cells uptake the greatest amount of amiodarone with an average of 2.38 and 2.60 pg per cell, respectively, and HeLa and Hek 293 have a significantly lower amount of amiodarone at 0.43 and 0.36 pg per cell, respectively. The amount of cell-associated drug for the ensemble population measurement (LC/MS/MS) is compared with the ToF-SIMS single cell data: a similar amount of drug was detected per cell for the NR8383, and HepG2 cells at a greater level than that for the HEK293 cells. However, the two techniques did not agree for the HeLa cells, and we postulate potential reasons for this.

Citation

Newman, C. F., Havelund, R., Passarelli, M. K., Marshall, P. S., Francis, I., West, A., Alexander, M. R., Gilmore, I. S., & Dollery, C. T. (2017). Intracellular drug uptake: a comparison of single cell measurements using ToF-SIMS imaging and quantification from cell populations with LC/MS/MS. Analytical Chemistry, 89(22), 11944-11953. https://doi.org/10.1021/acs.analchem.7b01436

• Journal Article Type: Article
• Acceptance Date: Oct 17, 2017
• Online Publication Date: Nov 8, 2017
• Publication Date: Nov 10, 2017
• Deposit Date: Dec 7, 2017
• Publicly Available Date: Nov 9, 2018
• Journal: Analytical Chemistry
• Print ISSN: 0003-2700
• Electronic ISSN: 1520-6882
• Publisher: American Chemical Society
• Peer Reviewed: Peer Reviewed
• Volume: 89
• Issue: 22
• Pages: 11944-11953
• DOI: https://doi.org/10.1021/acs.analchem.7b01436
• Public URL: https://nottingham-repository.worktribe.com/output/893746
• Publisher URL: https://doi.org/10.1021/acs.analchem.7b01436
• Additional Information: This material is excerpted from a work that was published in Analytical Chemistry, copyright American Chemical Society after peer review. To access the final edited and published work see https://doi.org/10.1021/acs.analchem.7b01436 see http://pubs.acs.org/page/policy/articlesonrequest/index.html]
• Contract Date: Dec 7, 2017