Quantitation of Cyclosporin A in Cell Culture Media by Differential Mobility Mass Spectrometry (DMS-MS/MS)

  • Amol Kafle
  • James Glick
  • Stephen L. CoyEmail author
  • Paul Vouros
Part of the Methods in Molecular Biology book series (MIMB, volume 2084)


Cell permeability is an important factor in determining the bioavailability of therapeutics that is usually measured by cell culture testing. The concentration of pharmaceutical in a medium such as Hank’s Balanced Salt Solution with HEPES organic buffer (HBSS-HEPES) is measured at a series of time points, making simplicity and high throughput of the analytical method important characteristics. We report an electrospray differential mobility spectrometry mass spectrometry method (nanoESI-DMS-MS) for the rapid determination of cyclosporin A (CsA, cyclosporine) concentration in such a buffer. DMS technology provides gas phase atmospheric pressure ion filtration for small-molecule bioanalytical methods that suppresses interfering ions and reduces chemical noise, without the use of chromatography. This allows simplified sample preparation, fast calibration curve development, and shortened analysis times. It has also been noted that the DMS prefilter can reduce contamination of the mass spectrometer by salts, thereby extending mass spectrometer system uptime.

In the application described here, DMS-MS/MS is applied to cyclosporine A (CsA) in cell medium. Sample preparation is limited to dilution with an ammonium acetate-methanol-water mobile phase and the addition of CsA-d4 internal standard. The isotope ratio data are obtained in DMS-MS MRM mode observing NH3 loss from the ammonium adduct of the two species. A calibration curve with high linearity (R2 = 0.998) is rapidly obtained with nearly zero intercept, while it was found that a liquid chromatography LC-MS method required a preliminary SPE step to obtain a linear calibration curve. The time for data acquisition in the DMS-MS MRM method with flow injection (FIA) or infusion introduction at ESI flow of 400 nL/min is typically 30 s leading to a cycle time of less than 1 min.

Key words

Cyclosporine Cyclosporine A Differential mobility spectrometry Mass spectrometry DMS DMS-MS Bioavailability Cell permeability 



Development of this method was supported by NIH: RO1 CA 069390-16 (Paul Vouros, PI).


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Amol Kafle
    • 1
  • James Glick
    • 2
  • Stephen L. Coy
    • 1
    Email author
  • Paul Vouros
    • 1
    • 3
  1. 1.Department of Chemistry and Chemical BiologyNortheastern UniversityBostonUSA
  2. 2.Novartis Institutes for BioMedical ResearchEast HanoverUSA
  3. 3.Barnett Institute of Chemical and Biological AnalysisNortheastern UniversityBostonUSA

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