A simple and sensitive assay for the quantitative analysis of rivastigmine and its metabolite NAP 226-90 in human EDTA plasma using coupled liquid chromatography and tandem mass spectrometry
Article first published online: 17 OCT 2006
Copyright © 2006 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 20, Issue 22, pages 3330–3336, 30 November 2006
How to Cite
Frankfort, S. V., Ouwehand, M., van Maanen, M. J., Rosing, H., Tulner, L. R. and Beijnen, J. H. (2006), A simple and sensitive assay for the quantitative analysis of rivastigmine and its metabolite NAP 226-90 in human EDTA plasma using coupled liquid chromatography and tandem mass spectrometry. Rapid Commun. Mass Spectrom., 20: 3330–3336. doi: 10.1002/rcm.2737
- Issue published online: 17 OCT 2006
- Article first published online: 17 OCT 2006
- Manuscript Revised: 11 SEP 2006
- Manuscript Accepted: 11 SEP 2006
- Manuscript Received: 6 JUL 2006
A sensitive and specific high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) assay for the determination of rivastigmine and its major metabolite NAP 226-90 is presented. A 100 µL plasma aliquot was spiked with a structural analogue of rivastigmine as internal standard (PKF214-976-AE-1) and proteins were precipitated by adding 200 µL of methanol. After centrifugation a volume of 100 µL of the clear supernatant was mixed with 100 µL of methanol/water (30:70, v/v) and volumes of 25 µL were injected onto the HPLC system. Separation was acquired on a 150 × 2.0 mm i.d. Gemini C18 column using a gradient system with 10 mM ammonium hydroxide and methanol. Detection was performed by using a turboionspray interface and positive ion multiple reaction monitoring by tandem mass spectrometry. The assay quantifies rivastigmine from 0.25 to 50 ng/mL and its metabolite NAP 226-90 from 0.50 to 25 ng/mL, using human plasma samples of 100 µL. Validation results demonstrate that rivastigmine and metabolite concentrations can be accurately and precisely quantified in human EDTA plasma. This assay is now used to support clinical pharmacologic studies with rivastigmine. Copyright © 2006 John Wiley & Sons, Ltd.