Simultaneous quantitative determination of cyclosporine A and its three main metabolites (AM1, AM4N and AM9) in human blood by liquid chromatography/mass spectrometry using a rapid sample processing method
Article first published online: 2 FEB 2006
Copyright © 2006 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 20, Issue 5, pages 733–740, 15 March 2006
How to Cite
Koseki, N., Nakashima, A., Nagae, Y. and Masuda, N. (2006), Simultaneous quantitative determination of cyclosporine A and its three main metabolites (AM1, AM4N and AM9) in human blood by liquid chromatography/mass spectrometry using a rapid sample processing method. Rapid Commun. Mass Spectrom., 20: 733–740. doi: 10.1002/rcm.2358
- Issue published online: 2 FEB 2006
- Article first published online: 2 FEB 2006
- Manuscript Accepted: 23 DEC 2005
- Manuscript Revised: 22 DEC 2005
- Manuscript Received: 1 JUL 2005
We have developed a sensitive and specific liquid chromatography/mass spectrometry (LC/MS) method for the simultaneous determination of cyclosporine A (CsA) and its three main metabolites (AM1, AM4N and AM9) in human blood. Following protein precipitation, supernatant was directly injected into the LC/MS system. Chromatographic separation was accomplished on a Symmetry C8 (4.6 × 75 mm, 3.5 µm) column with a linear gradient elution prior to detection by atmospheric pressure chemical ionization (APCI) MS using selected ion monitoring (SIM) in positive mode. This method can be applied to single mass equipment. The analytical range for each analyte was set at 1–2500 ng/mL using 100 µL of blood sample. The analytical method was fully validated according to FDA guidance. Intra-day mean accuracy and precision were 95.2–113.5% and 0.9–8.9%, respectively. Inter-day mean accuracy and precision were 95.8–107.0% and 1.5–10.7%, respectively. In blood all analytes were stable during three freeze/thaw cycles, for 24 h at room temperature and for 12 months at or below −15°C. Stability was also confirmed in processed samples for 24 h at 10°C and for 6 months at 4°C in methanol. In addition, we confirmed the method could avoid matrix effects from transplant subjects' samples. This LC/MS technique provided an excellent method for simultaneous quantitative determination of CsA and its three metabolites for evaluation of their pharmacokinetic profiles. Copyright © 2006 John Wiley & Sons, Ltd.