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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
Author(s) -
Frankfort Suzanne V.,
Ouwehand Mariët,
van Maanen Maria J.,
Rosing Hilde,
Tulner Linda R.,
Beijnen Jos H.
Publication year - 2006
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.2737
Subject(s) - chemistry , chromatography , metabolite , tandem mass spectrometry , rivastigmine , mass spectrometry , high performance liquid chromatography , liquid chromatography–mass spectrometry , biochemistry , medicine , dementia , disease , pathology , donepezil
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.

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